Difference between revisions of "Part:BBa K2259019"
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Amino 3'-glycosyl phosphotransferase (APH(3')) – a protein granting the resistance to aminoglycoside family antibiotics was split into two subunits [https://parts.igem.org/Part:BBa_K2259018 Alpha] and Beta (This part!) between amino acid residues 59 and 60 as introduced by Calvin M. Schmidt et al and [[Part:BBa_K1442031]]. | Amino 3'-glycosyl phosphotransferase (APH(3')) – a protein granting the resistance to aminoglycoside family antibiotics was split into two subunits [https://parts.igem.org/Part:BBa_K2259018 Alpha] and Beta (This part!) between amino acid residues 59 and 60 as introduced by Calvin M. Schmidt et al and [[Part:BBa_K1442031]]. | ||
− | This is an improved [[Part:BBa_K1442031]]. It has been properly split. | + | This is an improved [[Part:BBa_K1442031]] part. It has been properly split. |
We did not include the start codon at the 5' end of the gene as reports say it has an alternative start codon. | We did not include the start codon at the 5' end of the gene as reports say it has an alternative start codon. | ||
Revision as of 10:48, 1 November 2017
Beta-neomycin resistance protein
Split antibiotic resistance gene.
Amino 3'-glycosyl phosphotransferase (APH(3')) – a protein granting the resistance to aminoglycoside family antibiotics was split into two subunits Alpha and Beta (This part!) between amino acid residues 59 and 60 as introduced by Calvin M. Schmidt et al and Part:BBa_K1442031.
This is an improved Part:BBa_K1442031 part. It has been properly split. We did not include the start codon at the 5' end of the gene as reports say it has an alternative start codon.
If both alpha and beta DNA sequences are transcribed and translated in the cell, they can combine and form a fully functional antibiotic resistance protein.
This part is used in SynORI multi-plasmid framework selection system, in which up to 5 unique plasmid groups can be maintained in a single cell using only one antibiotic resistance protein.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 452
- 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 301
Illegal SapI.rc site found at 511
Contents
Introduction
Biology
Split antibiotic resistance
Aminoglycoside-3'-phosphotransferase (APH(3')), sometimes called aminoglycoside kinase, is an enzyme that catalyzes the addition of phosphoryl group from ATP to the 3'-hydroxyl group of a 4,6-disubstituted aminoglycoside, such as kanamycin, neomycin.
Aminoglycoside-3'-phosphotransferase gene was split by Calvin M. Schmidt et al to produce a protein that is enzymatically active only when a from alpha and beta subunits is formed
Usage with SynORI (Framework for multi-plasmid systems)
About SynORI
SynORI is a framework for multi-plasmid systems created by Vilnius-Lithuania 2017 which enables quick and easy workflow with multiple plasmids, while also allowing to freely pick and modulate copy number for every unique plasmid group! Read more about [http://2017.igem.org/Team:Vilnius-Lithuania SynORI here]!
Split antibiotic resistance in SynORI
Split antibiotic coding gene, together with toehold switches and their corresponding RNA triggers completes the dynamic SynORI selection system. The switches lock the translation of downstream split antibiotic genes and form an AND type gate genetic circuit which functions to stably maintain multiple plasmids in the SynORI collection.
SynORI selection gene circuits for multi-plasmid systems:
• 2 plasmids
Consisting of: : Two split antibiotic genes (part:BBa_K2259018 and part:BBa_K2259019)
• 3 plasmids
Consisting of: One Toehold (part:BBa_K2259014 or part:BBa_K2259015), one Trigger RNA (part:BBa_K2259016 or part:BBa_K2259017) and split neomycin antibiotic resistance genes (part:BBa_K2259018 and part:BBa_K2259019).
• 4 plasmids
Consisting of: Two Toeholds (part:BBa_K2259014 and part:BBa_K2259015), two Trigger RNAs (part:BBa_K2259016 and part:BBa_K2259017) and split neomycin antibiotic resistance genes (part:BBa_K2259018 and part:BBa_K2259019).
• 5 plasmids
Consisting of: Modified phage control system part:BBa_K2259044, two Toeholds (part:BBa_K2259014 and part:BBa_K2259015), two repressed Trigger RNAs (part:BBa_K2259042 and part:BBa_K2259043) and split neomycin antibiotic resistance genes (part:BBa_K2259018 and part:BBa_K2259019).
References
Stable Maintenance of Multiple Plasmids in E. coli Using a Single Selective Marker Calvin M. Schmidt, David L. Shis, Truong D. Nguyen-Huu, and Matthew R. Bennett ACS Synthetic Biology 2012 1 (10), 445-450 DOI: 10.1021/sb3000589