Bacillus subtilis

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Bacillus subtilis is gram-positive model organism. Thus, much is known about this organism. The genome of Bacillus subtilis strain 168 has been sequenced.

Although the current part collection for B. subtilis is small, many are now using B. subtilis as a candidate host for synthetic devices and systems. Please read more about the advantages and disadvantages of using B. subtilis as a chassis.

Plasmid backbones (?) Promoters (?) Ribosome binding sites (?)
Protein coding sequences (?) DNA parts (?)
Help: Want to know more about Bacillus subtilis? See the help pages for more information.

Plasmid backbone

BioBrickbackbonevspart.png

Plasmids are circular, double-stranded DNA molecules typically containing a few thousand base pairs that replicate within the cell independently of the chromosomal DNA. Plasmid DNA is easily purified from cells, manipulated using common lab techniques and incorporated into cells. Most BioBrick parts in the Registry are maintained and propagated on plasmids. Thus, construction of BioBrick parts, devices and systems usually requires working with plasmids.

Note: In the Registry, plasmids are made up of two distinct components:

  1. the BioBrick part, device or system that is located in the BioBrick cloning site, between (and excluding) the BioBrick prefix and suffix.
  2. the plasmid backbone which propagates the BioBrick part. The plasmid backbone is defined as the sequence beginning with the BioBrick suffix, including the replication origin and antibiotic resistance marker, and ending with the BioBrick prefix. [Note that the plasmid backbone itself can be composed of BioBrick parts.]

Many BioBrick parts in the Registry are maintained on more than one plasmid backbone!


The plasmids for B. Subtilis you can find in the registry are usually not built as standard registry plasmid so far. They come from third parties sources, that have been biobricked. Hence, these plasmid are often not silent. Cloning inside often works in different conditions than with standard E. coli plasmids. You may need to optimize again your ligation conditions.

As a particular drawback we noticed using replicative plasmids, is that they are often subject to recombination. We invite you to read in detail the experiment done and the peer review page before choosing a plasmid and a strategy.

There are 2 main kind of plasmids for B. Subtilis:

  • Replicative or also called episomal plasmids, are the same kind of plasmid with a replication origin and a resistance gene we are used to work with in E. Coli. These plasmids are often made to be multihost, so that they can be cloned using E. Coli and then the miniprep can be transformed into B. subtilis by starvation or electroporation. (Note that some replicative plasmids requires to be transformed into RecA+ E. Coli strains first before incorporating by starvation)
  • Integration plasmids is the most used technique by specialist in B. subtilis. They are E. Coli plasmids with two homologous sequence with two fragments of a chromosomal gene of B. Subtilis, and a resistance cassette inserted in between. If your brick is cloned in between these two homologous region, the vector act as a shuttle that integrate your construct into the chromosome.


Note that you should check your backbone and biobrick part sequence whether more than one side can be cut by same enzyme.You should not use if it has more than one same restriction enzyme site.

If you always see same bands in every gel images as different bp length from part page, you can try to transform your plasmid. Make sure that antibiotic you will use matches with which the backbone has..

And also if you see another band too, always at the same place and since first run. You can try transform your plasmid and check the viability when you exclude the possibility of contamination or any other things which can cause this problem in your lab. Another possibility could be that the part itself has already additional contaminated DNAs.



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NameDescriptionResistanceRepliconCopy
number
ChassisLength
BBa_I742123Multi-host vector pTG262 converted to BioBrick vectorCMulti-host Multi-host5564
BBa_J179000pBS1C: Bacillus subtilis vector, amyE integration, CAM-resistanceA(E. coli) C(B. subtilis)  B. subtilis + E. coli6105
BBa_J179001pBS2E: Bacillus subtilis vector, lacA integration, MLS resistanceA(E. coli) E(B. subtilis)  B. subtilis + E. coli6258
BBa_J179002pBs4S: Bacillus subtilis vector, thrC integration, spec resistanceA(E. coli) S(B. subtilis)pDG1731integrative in B. subtilisB. subtilis + E. coli4573
BBa_K090402B. subtilis Episomal Vector with Constitutive GFP     6337
BBa_K090403Gram-positive Shuttle Vector for Chromosomal Integration    5632
BBa_K1065204Efe+Bba_B0015 in BBa_K823024 (pXyl) A(E. coli) S(B. subtilis)  B. subtilis + E. coli6017
BBa_K1085014pSB1AC3-HySp: Integrational backbone into B. subtilis amyE locus with IPTG inducible promoter    7625
BBa_K1185004Integration vector for Bacillus subtilis derived from pSac-CmACmulti-host multi-host5210
BBa_K1364021Integrative plasmid for Bacillus subtilis (pSBbs4E)    5760
BBa_K818000Integration vector for Bacillus subtilis derived from pSac-CmACMulti host Multi host5223
BBa_K823021pSBBs1C-lacZ (lacZ reporter vector for B. subtilis)A(E. coli) C(B. subtilis)pAC6integrative in B. subtilisE.coli and B. subtilis9792
BBa_K823022pSBBs4S: Empty backbone for integration into Bacillus subtilis thrC locusA(E. coli) S(B. subtilis)pDG1731integrative in B. subtilisB. subtilis + E. coli4573
BBa_K823023pSBBs1C: Empty backbone for integration into Bacillus subtilis amyE locusA(E. coli) C(B. subtilis)  B. subtilis + E. coli6105
BBa_K823024pSBBs4S-Pxyl: Integrative expression vector for Bacillus subtilisA(E. coli) S(B. subtilis)  B. subtilis + E. coli4794
BBa_K823025pSBBs3C-luxABCDE (lux reporter vector for B.subtilis)A(E. coli) + C(B. subtilis)pAH328integrative in B. subtilisB. subtilis + E. coli10640
BBa_K823027pSBBs2E: Empty backbone for integration into Bacillus subtilis lacA locusA(E. coli) E(B. subtilis)  B. subtilis + E. coli6193

Constitutive promoters

ConstitutivePromoter.png

The promoters here are B. subtilis promoters that are constitutive meaning that the activity of these promoters should only be regulated by the levels of RNA polymerase and the appropriate σ factor.

The sequence of these promoter are adapted to the σ factor of B. subtilis. However, some of these promoter also works in E. coli. Generally speaking, standard E. coli promoters don't work (or are very weak) in B. subtilis strains, whereas the contrary generally works. However, it doesn't mean that the efficiency will be the same in both strains. The pVeg promoter, for instance, works fine at a high level of expression in both E. coli and B. subtilis strains - Contribution from User: Cyrpaut (31 October 2011)

Constitutive B. subtilis σA promoters

This section lists promoters that are recognized by B. subtilis σA RNAP. σA is the major B. subtilis sigma factor so there should be RNAP present to transcribe these promoters under most growth conditions (although maximally during exponential growth).


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NameDescriptionPromoter SequencePositive
Regulators
Negative
Regulators
LengthDocStatus
BBa_K143012Promoter veg a constitutive promoter for B. subtilis . . . aaaaatgggctcgtgttgtacaataaatgt976561In stock
BBa_K143013Promoter 43 a constitutive promoter for B. subtilis . . . aaaaaaagcgcgcgattatgtaaaatataa5611873Not in stock
BBa_K780003Strong constitutive promoter for Bacillus subtilis . . . aattgcagtaggcatgacaaaatggactca361559It's complicated
BBa_K823000PliaG . . . caagcttttcctttataatagaatgaatga1217765In stock
BBa_K823002PlepA . . . tctaagctagtgtattttgcgtttaatagt1577245In stock
BBa_K823003Pveg . . . aatgggctcgtgttgtacaataaatgtagt23714126In stock

Constitutive B. subtilis σB promoters

This section lists promoters that are recognized by B. subtilis σB RNAP. σB is the major stationary phase E. coli sigma factor. Use these promoters when you want high promoter activity during stationary phase or during starvation.


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NameDescriptionPromoter SequencePositive
Regulators
Negative
Regulators
LengthDocStatus
BBa_K143010Promoter ctc for B. subtilis . . . atccttatcgttatgggtattgtttgtaat564297Not in stock
BBa_K143011Promoter gsiB for B. subtilis . . . taaaagaattgtgagcgggaatacaacaac382765Not in stock
BBa_K143013Promoter 43 a constitutive promoter for B. subtilis . . . aaaaaaagcgcgcgattatgtaaaatataa5611873Not in stock

Positively regulated promoters

PositivePromoter.png

The B. subtilis promoters of this section is the ones that are said to be positively regulated. It means that meaning their expression level increase with the help of another third party protein called transcription activator (This category exclude the sigma factor protein itself). With the appropriate protein, you would be able to increase the activity of your promoter. Please read the description and characterization of each parts for more details.

Positively regulated B. subtilis σA promoters

This section lists the promoters recognized by B. subtilis σA RNA polymerase sub-unit. σA is the major B. subtilis sigma factor that is present under most growth conditions (but maximal during exponential growth phase).


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NameDescriptionPromoter SequencePositive
Regulators
Negative
Regulators
LengthDocStatus
BBa_K090504Gram-Positive Strong Constitutive Promoter . . . acatgggaaaactgtatgtatttgatcctc2392275It's complicated

Positively regulated B. subtilis σB promoters

This section lists promoters that are recognized by B. subtilis σB RNA polymerase. σB is the polymerase subunit that is the most present during the stationary growth phase. You can use these promoters if you want your construct to be mostly expressed during stationary growth phase or under starvation conditions.


There are no parts for this table

Repressible promoters

NegativePromoter.png

The B. subtilis promoters of this section are said negativly regulated promoters, because they can be repressed by the expression of a third party protein. The inhibition can be released by the addition of a molecule, like for the LacI E. coli promoter.

In the following biobricks, the proposed promoters are build with the fusion of one or several operons with a σA type contitutive promoter.


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NameDescriptionPromoter SequencePositive
Regulators
Negative
Regulators
LengthDocStatus
BBa_K090501Gram-Positive IPTG-Inducible Promoter . . . tggaattgtgagcggataacaattaagctt1072054It's complicated
BBa_K143014Promoter Xyl for B.subtilis . . . agtttgtttaaacaacaaactaataggtga822112Not in stock
BBa_K143015Promoter hyper-spank for B. subtilis . . . aatgtgtgtaattgtgagcggataacaatt1012527Not in stock


In the future, we may also find promoters builded with the σB promoter.


There are no parts for this table

Ribosome binding sites

B. subtilis have the same system of RBS than E. coli to drive its transcription. Hoever, the RBS sequences are not always compatible since B. subtilis ribsomomal RNA (rRNA) is slightly different in sequence compare to E. coli.

In this list, you can find the most common RBSs usually use in B. subtilis. They are classified by strengh in a qualitative manner, since no strengh reference ranking have been established for B. subtilis yet. It is not obvious to transport directly the system for E. coli since we need to have a promoter that is exactly the same strengh as an E. coli one to make both scales comparable between the organism.

We can also notice that E. coli RBSs usually doesn't works for B. subtilis, but it is not rare to find B. subtilis RBSs that works fine in E. coli (the SpoVg RBS for instance).

Please go in the description of each parts for more information.


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NameSequenceDescriptionRelative
Strength
Predicted
Strength
BBa_K090505aaaggaggtgt''Bacillus subtilis'' consensus RBS  
BBa_K090506agaggtggtgt''Bacillus subtilis'' weak RBS  
BBa_K104003agggggccgRBS_spaR  
BBa_K104005ggggcgttgRBS_spaK  
BBa_K104009agaggaggRBS_gfp  
BBa_K1351028aaggagggataLMU Bacillus RBS collection 1  
BBa_K1351029agaggaggataLMU Bacillus RBS collection 2  
BBa_K1351030aaggagagataLMU Bacillus RBS collection 3  
BBa_K1351031aggagaggataLMU Bacillus RBS collection 4  
BBa_K1351032aagaggagataLMU Bacillus RBS collection 5  
BBa_K1351033agaaagggataLMU Bacillus RBS collection 6  
BBa_K1351034aagaagagataLMU Bacillus RBS collection 7  
BBa_K143020taaaggaggaaGsiB ribosome binding site (RBS) for B. subtilis  
BBa_K143021aaaggtggtgaaSpoVG ribosome binding site (RBS) for B. subtilis  
BBa_K2586010 . . . tgctacaattactagaggtttaaaggaggaPalf4-RBS: Palf4 linked to a RBS  
BBa_K3831005gtaataagtaggttaggagaggSynthetic RBS_a  
BBa_K3831006 . . . actacacgacaattaaagaaggtattttttSynthetic RBS_b  
BBa_K3831007ggtggaaaggaggtgatcgacNative RBS R2  
BBa_K3831008ggtgggaaggagggggttcgacRBS R6  
BBa_K3831009gggatagacccagggggaggtttttttSynthetic RBS_c  
BBa_K3831014gctcttaaggaggattttagaNative RBS R1  
BBa_K3831018gattaactaataaggaggacaaacNative RBS R0  
BBa_K3831020ggtgggaaggaggacattcgacArtificial RBS R3  
BBa_K3831047aaaaaaacctccccctgggtctatcccSynthetic RBS_c - reversed  
BBa_K3831049gtcgaaccccctccttcccaccRBS R6 - reversed  
BBa_K3831051gtcgatcacctcctttccaccRBS R2 - reversed  
BBa_K3831053 . . . aaataccttctttaattgtcgtgtagtaatSynthetic RBS_b - reversed  
BBa_K3831055cctctcctaacctacttattacSynthetic RBS_a - reversed  
BBa_K4382009aaggaggaaggatcaRBS for Bacillus Subtilis  
BBa_K780001atattaagaggaggagStrong RBS for Bacillus Subtilis  
BBa_K780002agagaacaaggaggggStrong RBS for Bacillus Subtilis  

Protein coding sequences

Protein coding sequences are DNA sequences that are transcribed into mRNA and in which the corresponding mRNA molecules are translated into a polypeptide chain. Every three nucleotides, termed a codon, in a protein coding sequence encodes 1 amino acid in the polypeptide chain. In some cases, different chassis may either map a given codon to a different sequence or may use different codons more or less frequently. Therefore some protein coding sequences may be optimized for use in a particular chassis.

In the Registry, protein coding sequences begin with a start codon (usually ATG) and end with a stop codon (usually with a double stop codon TAA TAA). Protein coding sequences are often abbreviated with the acronym CDS.

Although protein coding sequences are often considered to be basic parts, in fact proteins coding sequences can themselves be composed of one or more regions, called protein domains. Thus, a protein coding sequence could either be entered as a basic part or as a composite part of two or more protein domains.

  1. The N-terminal domain of a protein coding sequence is special in a number of ways. First, it always contains a start codon, spaced at an appropriate distance from a ribosomal binding site. Second, many coding regions have special features at the N terminus, such as protein export tags and lipoprotein cleavage and attachment tags. These occur at the beginning of a coding region, and therefore are termed Head domains.
  2. A protein domain is a sequence of amino acids which fold relatively independently and which are evolutionarily shuffled as a unit among different protein coding regions. The DNA sequence of such domains must maintain in-frame translation, and thus is a multiple of three bases. Since these protein domains are within a protein coding sequence, they are called Internal domains. Certain Internal domains have particular functions in protein cleavage or splicing and are termed Special Internal domains.
  3. Similarly, the C-terminal domain of a protein is special, containing at least a stop codon. Other special features, such as degradation tags, are also required to be at the extreme C-terminus. Again, these domains cannot function when internal to a coding region, and are termed Tail domains.

For more details on protein domains including how to assemble protein domains into protein coding sequences, please see Protein domains.


Protein coding sequences should be as follows

GAATTC GCGGCCGC T TCTAG [ATG ... TAA TAA] T ACTAGT A GCGGCCG CTGCAG


Note of the codon availability in B. subtilis: B. subtilis is more codon tolerent than E. coli, which means that you can generally express any protein from E. coli into B. subtilis without any problem. However, some proteins are said to be for subtilis for two reasons: One is because they comes from B. subtilis genome, (which means that you may have trouble in expressing them in E. coli) or because they are made to trigger a certain effect in B. subtilis (for instance kill it).



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NameProteinDescriptionTagDirectionUniProtKEGGLengthStatus
BBa_E0040GFPmut3b green fluorescent protein derived from jellyfish Aequeora victoria wild-type GFP (SwissProt: P42212NoneForward  720In stock
BBa_E1010mRFP1 **highly** engineered mutant of red fluorescent protein from Discosoma striata (coral)NoneForward  706In stock
BBa_K104004 spaR CDS (Coding sequence)  663Not in stock
BBa_K104006 spaK CDS (Coding Sequence)  1182Not in stock
BBa_K1085000 RBS Start-codon Strep-tag SpiderSilkSubunitE1  268In stock
BBa_K1085001 RBS Start-codon SpiderSilkSubunitE1   226In stock
BBa_K1085002 SpiderSilkSubunitE2  210In stock
BBa_K1085003 SpiderSilkSubunitE2 Strep-tag   252In stock
BBa_K1085004 RBS Start-codon SpiderSilkSubunitN1 Strep-tag  199In stock
BBa_K1085005 RBS Start-codon SpiderSilkSubunitN1   157In stock
BBa_K1085006 SpiderSilkSubunitN2   129In stock
BBa_K1085007 SpiderSilkSubunitN2 Strep-tag  171In stock
BBa_K1085010 RBS Start-codon EstA Strep-tag SpiderSilkSubunitE1   907It's complicated
BBa_K1085012 RBS Start-codon EstA Strep-tag SpiderSilkSubunitN1   838In stock
BBa_K1085029 FliZ Strep-tag SpiderSilkSubunitE1  339In stock
BBa_K1085034 MotB Strep-tag SilkSubunitN1  309In stock
BBa_K1085050 des knock out  3604Not in stock
BBa_K1122000 SinR transcription factor  333It's complicated
BBa_K1122666 Ferric uptake repressor  450It's complicated
BBa_K1351000 beta-Neurexin-derived peptide binding to S. aureus adhesin SdrC  33In stock
BBa_K1351001 Contactin-4-derived peptide binding to NADH Oxidase of S. pneumoniae   42In stock
BBa_K1351002 Chondroitin 4 sulfotransferase-derived peptide binding to NADH Oxidase of S. pneumoniae   39In stock
BBa_K1351003 Laminin-derived peptide binding to NADH Oxidase of S. pneumoniae   27In stock
BBa_K1351006 CWB domain of B. subtilis major autolysin LytC  999In stock
BBa_K1351007 CWB domain of B. subtilis minor autolysin LytE  621In stock
BBa_K1351008 Signal peptide of B. subtilis endonuclease YhcR  138In stock
BBa_K1351010 Cell wall-anchoring domain of B. subtilis endonuclease YhcR  402In stock
BBa_K1351011 Bacillus subtilis sortase YhcS  597In stock
BBa_K1351012 spaS Subtilin (antimicrobial peptide, Freiburg standard)  168In stock
BBa_K1351013 spaS Subtilin (antimicrobial peptide)  171In stock
BBa_K1351017 SdpI with RBS: Immunity against the cannibalsim toxin sdpC of B. subtilis  333In stock
BBa_K1351021 Monomeric Red Fluorescent Protein from Discosoma striata  675In stock
BBa_K1351026 Signal peptide of B. subtilis major autolysin LytC  75In stock
BBa_K1351027 Signal peptide of B. subtilis minor autolysin LytE  75In stock
BBa_K1362461mRFP1**highly** engineered mutant of red fluorescent protein from Discosoma striata (coral) with barcode NoneForward  706Not in stock
BBa_K1399000mRFP1RFP from Discosoma striata (coral) with AAV-ssrA degradation tagSsrA-AAVForward  714In stock
BBa_K1399001mRFP1RFP from Discosoma striata (coral) with LVA-ssrA degradation tagSsrA-LVA degradation tagForward  714In stock
BBa_K1399002mRFP1RFP from Discosoma striata (coral) with LAA-ssrA degradation tag (wt)SsrA-LAA degradation tagForward  714In stock
BBa_K1399003mRFP1RFP from Discosoma striata (coral) with DAS-ssrA degradation tagSsrA-DAS degradation tagForward  714In stock
BBa_K1399004GFPmut3bGFP (mut3b) with LVA-ssrA degradation tagSsrA-LVA degradation tagForward  753In stock
BBa_K1399005GFPmut3bGFP (mut3b) with AAV-ssrA degradation tagSsrA-AAV degradation tagForward  753In stock
BBa_K1399006GFPmut3bGFP (mut3b) with LAA-ssrA degradation tagSsrA-LAA degradation tagForward  753In stock
BBa_K1399007GFPmut3bGFP (mut3b) with SsrA-DAS+2 degradation tagSsrA-DAS+2 degradation tagForward  759In stock
BBa_K1399008GFPmut3bGFP (mut3b) with DAS-ssrA degradation tagSsrA-DAS degradation tagForward  753In stock
BBa_K143032 EpsE Molecular Clutch Gene of B. subtilis  840In stock
BBa_K143034 LipA-EAK16-II Fusion Protein  156It's complicated
BBa_K143035 LipA-Human Elastin (EP20-24-24) Fusion Protein  705It's complicated
BBa_K143036 Xylose operon regulatory protein  1056It's complicated
BBa_K143037 YtvA Blue Light Receptor for B.subtilis  789It's complicated
BBa_K143038 SacB-EAK16-II Fusion Protein  150It's complicated
BBa_K143039 SacB-Human Elastin (EP20-24-24) Fusion Protein  699It's complicated
BBa_K143063 Xylose operon repressor protein - Terminator  1193Not in stock
BBa_K1701000 GolB  195In stock
BBa_K1701001 PbrR  438It's complicated
BBa_K1701004 SUP  267It's complicated
BBa_K1701005 TasA  786It's complicated
BBa_K1701006 CotC  201It's complicated
BBa_K174012 sleB, Bacillus subtilis germination gene with RBS  932It's complicated
BBa_K2057002 Alginate lyase from Bacillus thuringiensis   No part sequence
BBa_K2273004 Epr signal peptide of B. subtilis minor extracellular serine protease  81It's complicated
BBa_K2273005 FliL signal peptide of B. subtilis flagellar protein  69It's complicated
BBa_K2273006 FliZ signal peptide of B. subtilis flagellar biosynthetic protein  81It's complicated
BBa_K2273007 GlpQ signal peptide of B. subtilis glycerolphosphate diester phosphodiesterase  78It's complicated
BBa_K2273008 LipA signal peptide of B. subtilis lipoic acid synthase  96It's complicated
BBa_K2273009 LytB signal peptide of B. subtilis modifier protein of major autolysin LytC  75It's complicated
BBa_K2273010 LytC signal peptide of B. subtilis N-acetylmuramoyl-L-alanine amidase  72It's complicated
BBa_K2273011 LytD signal peptide of B. subtilis peptidoglycan N-acetylglucosaminidase  81It's complicated
BBa_K2273015 Minimal SpyCatcher codon adapted for Bacillus subtilis  249It's complicated
BBa_K2273023 AmyE signal peptide of B. subtilis alpha-amylase  99It's complicated
BBa_K2273024 AspB signal peptide of B. subtilis aspartate transaminase  66It's complicated
BBa_K2273025 BglS signal peptide of B. subtilis endo-beta-1,3-1,4 glucanase  84It's complicated
BBa_K2273026  CccA signal peptide of B. subtilis cytochrome c550  81It's complicated
BBa_K2273027 Bpr signal peptide of B. subtilis bacillopeptidase F  90It's complicated
BBa_K2273028 Csn signal peptide of B. subtilis chitosanase  105It's complicated
BBa_K2273029 DacB signal peptide of B. subtilis sporulation-specific penicillin-binding protein  81It's complicated
BBa_K2273030 DacF signal peptide of B. subtilis penicillin-binding protein I  69It's complicated
BBa_K2273031 DltD signal peptide of B. subtilis Protein DltD  63It's complicated
BBa_K2273032 CitH signal peptide of B. subtilis malate dehydrogenase  96It's complicated
BBa_K2273043 LytR signal peptide of B. subtilis phosphotransferase  117It's complicated
BBa_K2273051 PbpX signal peptide of B. subtilis penicillin-binding protein X  117It's complicated
BBa_K2273052 Pel signal peptide of B. subtilis pectate lyase C  63It's complicated
BBa_K2273053 PelB signal peptide of B. subtilis pectate lyase  72It's complicated
BBa_K2273054 PenP signal peptide of B. subtilis beta-lactamase  108It's complicated
BBa_K2273055 PhoA signal peptide of B. subtilis alkaline phosphatase A  93It's complicated
BBa_K2273056 PhoB signal peptide of B. subtilis alkaline phosphatase III  96It's complicated
BBa_K2273057 PhrA signal peptide of B. subtilis Phosphatase RapA inhibitor  75It's complicated
BBa_K2273058 PhrC signal peptide of B. subtilis Phosphatase RapC inhibitor  81It's complicated
BBa_K2273059 PhrF signal peptide of B. subtilis Phosphatase RapF inhibitor  72It's complicated
BBa_K2273060 PhrG signal peptide of B. subtilis Phosphatase RapG inhibitor  63It's complicated
BBa_K2273061 PhrK signal peptide of B. subtilis Phosphatase RapK inhibitor  57It's complicated
BBa_K2273062 RpmGB signal peptide of B. subtilis 50S ribosomal protein L33 2  78It's complicated
BBa_K2273063 SacB signal peptide of B. subtilis levansucrase  87It's complicated
BBa_K2273064 SacC signal peptide of B. subtilis levanase  72It's complicated
BBa_K2273065 SleB signal peptide of B. subtilis spore cortex-lytic enzyme  93It's complicated
BBa_K2273066 SpoIID signal peptide of B. subtilis lytic transglycosylase  102It's complicated
BBa_K2273067 SpoIIP signal peptide of B. subtilis cell wall hydrolase  117It's complicated
BBa_K2273068 SpoIIQ signal peptide of B. subtilis stage II sporulation protein Q  120It's complicated
BBa_K2273069 SpoIIR signal peptide of B. subtilis stage II sporulation protein R  60It's complicated
BBa_K2273070 TyrA signal peptide of B. subtilis prephenate dehydrogenase  69It's complicated
BBa_K2273071 Vpr signal peptide of B. subtilis minor extracellular serine protease  93It's complicated
BBa_K2273072 WapA signal peptide of B. subtilis cell wall-associated protein precursor  96It's complicated
BBa_K2273073 YbbC signal peptide of B. subtilis unknown product  69It's complicated
BBa_K2273074 YbbE signal peptide of B. subtilis N-acetylmuramyl-L-alanine amidase  75It's complicated
BBa_K2273075 YbbR signal peptide of B. subtilis CdaA regulatory protein CdaR  84It's complicated
BBa_K2273076 YbdG signal peptide of B. subtilis Uncharacterized protein YbdG  75It's complicated
BBa_K2273077 YbdN signal peptide of B. subtilis Uncharacterized protein YbdN  75It's complicated
BBa_K2273078 YbfO signal peptide of B. subtilis Putative hydrolase YbfO  84It's complicated
BBa_K2273079 YbxI signal peptide of B. subtilis Probable beta-lactamase YbxI  69It's complicated
BBa_K2273080 YdbK signal peptide of B. subtilis Uncharacterized membrane protein YdbK  99It's complicated
BBa_K2273081 YdhT signal peptide of B. subtilis Mannan endo-1,4-beta-mannosidase  63It's complicated
BBa_K2273092 YjcN signal peptide of B. subtilis uncharacterized protein YjcN  102It's complicated
BBa_K2273093 YjdB signal peptide of B. subtilis uncharacterized protein YjdB  78It's complicated
BBa_K2273094 YjfA signal peptide of B. subtilis uncharacterized protein YjfA  84It's complicated
BBa_K2273095 YjiA signal peptide of B. subtilis uncharacterized protein YjiA  78It's complicated
BBa_K2273096 YkoJ signal peptide of B. subtilis uncharacterized protein YkoJ  78It's complicated
BBa_K2273097 YkvV signal peptide of B. subtilis sporulation thiol-disulfide oxidoreductase A  78It's complicated
BBa_K2273098 YkwD signal peptide of B. subtilis uncharacterized protein YkwD  78It's complicated
BBa_K2273099 YlaE signal peptide of B. subtilis uncharacterized protein YlaE  93It's complicated
BBa_K2273100 YlbL signal peptide of B. subtilis uncharacterized protein YlbL  105It's complicated
BBa_K2273101 YlqB signal peptide of B. subtilis inhibitor of entry into sporulation  81It's complicated
BBa_K2273102 YlxF signal peptide of B. subtilis FlaA locus 22.9 kDa protein  111It's complicated
BBa_K2273103 AmyEEV version of B. subtilis alpha-amylase without its native signal peptide  1881It's complicated
BBa_K2273104 AmyERFC25 version of B. subtilis alpha-amylase without its native signal peptide  1881It's complicated
BBa_K2586001 GltT: glutamate and glyphosate transporter in B. subtilis  1290It's complicated
BBa_K2586002 GltP: glutamate and glyphosate uptake transporter in Bacillus subtilis  1245It's complicated
BBa_K2586003 aroE: 3-phosphoshikimate 1-carboxyvinyltransferase  1287It's complicated
BBa_K2586007 aroA: 3-phosphoshikimate 1-carboxyvinyltransferase  1284It's complicated
BBa_K2586019 GAT: Glyphosate N-Acetyltransferase  440It's complicated
BBa_K2660000 Penetratin, cell-penetrating peptide  51Not in stock
BBa_K2660003 mCherry codon optimized for Lactococcus lactis  724Not in stock
BBa_K2660006 N-Cas9  3459It's complicated
BBa_K2660007 C-Cas9  648It's complicated
BBa_K2660008 nMag  450It's complicated
BBa_K2660009 pMag  450It's complicated
BBa_K294055GFPmut3bGFP RFP HybridNone  720In stock
BBa_K302001 yneA coding sequence  321Not in stock
BBa_K302006 spaI  498Not in stock
BBa_K302007 spaF  744Not in stock
BBa_K302008 spaE  756Not in stock
BBa_K302010 sfp  678Not in stock
BBa_K302011 swrA  432Not in stock
BBa_K302033 mazF  339In stock
BBa_K3128008mRFP1Engineered mutant of red fluorescent protein with RFC21 restriction sitesNoneForward  690Not in stock
BBa_K3590005 SacB_v1  87It's complicated
BBa_K3590006 SacB signal peptide of Bacillus subtilis levansucrase version 2  87Not in stock
BBa_K3590013 Cellulose binding module  495Not in stock
BBa_K3590014 Cohesin derived from Acetivibrio cellulolyticus  438Not in stock
BBa_K3590015 Dockerin derived from Acetivibrio cellulolyticus  228Not in stock
BBa_K3590016 Cohesin derived from Bacteroides cellulosolvens  450Not in stock
BBa_K3590017 Dockerin derived from Bacteroides cellulosolvens  243Not in stock
BBa_K3590018 Cohesin derived from Clostridium thermocellum  435Not in stock
BBa_K3590019 Dockerin derived from Clostridium thermocellum  219Not in stock
BBa_K3590020 3x LysM domain derived from Bacillus subtilis LytE version 1  621Not in stock
BBa_K3590021 3x LysM domain derived from Bacillus subtilis LytE version 2  621Not in stock
BBa_K3590022 Lysozyme derived from Bacillus licheniformis  951Not in stock
BBa_K3590024 Microvirin  324Not in stock
BBa_K3590025 MlrA  1008Not in stock
BBa_K3590026 MlrB  1623Not in stock
BBa_K3590027 MlrC  1584Not in stock
BBa_K3590028 Linker 1 version 1  87Not in stock
BBa_K3590029 Linker 1 version 2  87Not in stock
BBa_K3590030 Linker 1 version 3  87Not in stock
BBa_K3590031 Linker 1 version 4  87Not in stock
BBa_K3590032 Linker 2 version 1  105Not in stock
BBa_K3590033 Linker 2 version 2  105Not in stock
BBa_K3590034 Linker in front of a dockerin version 1  21Not in stock
BBa_K3590035 Linker in front of a dockerin version 2  21Not in stock
BBa_K3831000 BMC - pduA  285 
BBa_K3831001 BMC - pduB  783 
BBa_K3831002 BMC - pduJ  276 
BBa_K3831003 BMC - pduK  696 
BBa_K3831004 BMC - pduN  270 
BBa_K3831013 Ggggs linker  15 
BBa_K3831016 sfGFP  714 
BBa_K3831017 Localization tag for encapsulation in BMC  24 
BBa_K3831019 Degradation tag  45 
BBa_K3831021 mScarlet-I  696 
BBa_K3831024 cI repressor  711 
BBa_K3831029 Green fluorescent protein  714 
BBa_K3831042 GFP  714 
BBa_K3831046 PduN - reverse complement  270 
BBa_K3831048 PduK - reverse complement  696 
BBa_K3831050 PduJ - reverse complement  276 
BBa_K3831052 PduB - reverse complement  783 
BBa_K3831054 PduA - reverse complement  285 
BBa_K4202000 mazE(codon optimization for Bacillus subtilis)  249 
BBa_K4202001 mazF(codon optimization for Bacillus subtilis)  421 
BBa_K4382000CglTBeta-glucosidase  1353 
BBa_K4382001EG5C-1An endoglucanase, EG5C-1  996 
BBa_K4382002DyP1BDye decolourising peroxidase (DyP1B)  888 
BBa_K4382003xynBBeta-xylosidase (XynB)  1602 
BBa_K4382004 Glucuronoxylan depolymerase - Xylanase C  1269 
BBa_K4382005XynDArabinofuranoside - XynD  1542 
BBa_K4382006PelB-B2Endo Pectin Lyase - PelB-B2  1065Not in stock
BBa_K4382007PelAPectin lyase - PelA  1263 
BBa_K4382008PmePectin Methyl Esterase (Pme)  1191 
BBa_K4382010xynAAn endoxylanase - XynA  642 
BBa_K802006 Surfactin generator for B. subtilis  787Not in stock
BBa_K802007 Biofilm repressor for B. subtilis strains  304It's complicated
BBa_K802008 Transcriptional Bacillus regulator lacI  1193Not in stock
BBa_K802009 Sufactin generator and biofilm repressor for B. subtilis  2300It's complicated
BBa_K823020catchloramphenicol acetyltransferase (cat)  645It's complicated
BBa_K823031 cotZ: B. subtilis spore crust protein (-2aa)  435It's complicated
BBa_K823032 cotZ: B. subtilis spore crust protein  441It's complicated
BBa_K863121GFPmut3bgreen fluorescent protein derived from jellyfish Aequeora victoria wild-type GFP (His-tag)His6  750Not in stock


DNA parts for building integration vector

Any E. coli plasmid can be converted into an integration vector if one add two pieces of a a gene sequence that is homologous to a gene into B. subtilis genes, and an antibiotic resistance cassette or other screening method.

IntegrationAmyE.PNG
The AmyE integration DNA parts (BBa_K143001 and BBa_K143002) are two parts that can be added to the 5' and 3' ends of a construct to allow integration into the B. subtilis genome. These parts have been successfully used within the parts BBa_K143079 and BBa_K143082 for integration. Integrated synthetic biological systems offer better genetic stability and more regulated copy number than plasmid-borne systems. For more information about these parts, please see [http://2008.igem.org/Team:Imperial_College 2008 Imperial College iGEM team wiki.]


More...
NameDescriptionSequenceLength
BBa_K1430015 Integration Sequence for the amyE locus of B. subtilis . . . aacacacaaattaaaaactggtctgatcga522
BBa_K1430023 Integration Sequence for the amyE locus of B. subtilis . . . tcgggcttaagcggttctcttccccattga1002
BBa_K3831010Spacer_0 . . . cttactctgttgaaaacgaatagataggtt40
BBa_K3831011Spacer_1tgctcgtagtttacc15
BBa_K3831012Spacer_5 . . . agaatagtcaatcttcggaaatcccaggtg40
BBa_K3831015Spacer_7taataaaaggtcccg15
BBa_K3831023Spacer 03aaggaacggttattt15
BBa_K3831026Spacer 02agattactactgata15
BBa_K3831027Spacer 06ccgattctgagacgg15
BBa_K3831028Spacer 04 . . . aatacaggacccgaatcgtttcagttgcct40
BBa_K3831038Spacer_SP1 . . . ttaccacggatacagacagtgataatctta40

References

Given the number of available articles on B. subtilis, we only include some review articles here.

<biblio>

  1. Earl pmid=18467096
  2. Pavlendova pmid=18450217
  3. Sonenshein pmid=17982469
  4. Lopez pmid=17981078
  5. Aguilar pmid=17977783
  6. Irnov pmid=17381303

</biblio>