Difference between revisions of "Part:BBa K2842680"

Revision as of 21:44, 17 October 2018

Intein Monomer 1: RFP reporter flanked with orthogonal inteins

Intein Monomer 1
Function	Standardised blue-white screening
Use in	E. coli cells
Chassis Tested	DH5α cells, BL21* cells
Abstraction Hierarchy	Composite Device
Related Device	BBa_K2842690
RFC standard	RFC10,RFC12,RFC21,RFC23 & RFC25 compatible
Backbone	pSB1C3
Submitted by	[http://2018.igem.org/Team:UCL UCL iGEM 2018]

Figure 1: BsaI digestions

(1) BsaI digested Intein Passenger BBa_K2842669
(2) BsaI digested RFP inteins BBa_K2842680
(3) BsaI digested GFP inteins BBa_K2842690
*edited to show relevant bands

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI site found at 1165
Illegal BsaI.rc site found at 28
Illegal SapI site found at 903
Illegal SapI.rc site found at 213

Functional Parameters

Protein data table for BioBrick BBa_ automatically created by the BioBrick-AutoAnnotator version 1.0

Nucleotide sequence in RFC 10: (underlined part encodes the protein)
GCTTCTACAAACGCGGCTTCTTCCAAAGAGACCTAATACGACTCACTATAGGGGTTGTGAGCGGATAACAACCCAAGACAAGGAGGAGTACCAATGATCAAG ... CGCTTGGCT
TAAGTGACAGTTGAAAAGCGAAAAAAAAACCCCGCCCCTGACAGGGCGGGGTTTTTTTTGGTCTCAACGGACGACGCCGGTTACTACATTGA
ORF from nucleotide position 94 to 1032 (excluding stop-codon)

Amino acid sequence: (RFC 25 scars in shown in bold, other sequence features underlined; both given below)

1
101
201
301

MIKIATRKYLGKQNVYDIGVERDHNFALKNGFIASNCYNGGRASMASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAKLKVTKGGPLPF
AWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGA
LKGEIKMRLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHSTGAGSSSESGSWSHPQFEKAEYCVYGDTMVETED
GKIKIEDLYKRLA*

Sequence features: (with their position in the amino acid sequence, see the list of supported features)

Strep-tag II:

278 to 285

Amino acid composition:

Ala (A)	18 (5.8%)
Arg (R)	13 (4.2%)
Asn (N)	9 (2.9%)
Asp (D)	19 (6.1%)

Cys (C)	2 (0.6%)
Gln (Q)	10 (3.2%)
Glu (E)	29 (9.3%)
Gly (G)	32 (10.2%)

His (H)	7 (2.2%)
Ile (I)	15 (4.8%)
Leu (L)	16 (5.1%)
Lys (K)	30 (9.6%)

Met (M)	11 (3.5%)
Phe (F)	13 (4.2%)
Pro (P)	13 (4.2%)
Ser (S)	20 (6.4%)

Thr (T)	17 (5.4%)
Trp (W)	4 (1.3%)
Tyr (Y)	17 (5.4%)
Val (V)	18 (5.8%)

Amino acid counting

	Total number:	313
	Positively charged (Arg+Lys):	43 (13.7%)
	Negatively charged (Asp+Glu):	48 (15.3%)
	Aromatic (Phe+His+Try+Tyr):	41 (13.1%)

Biochemical parameters

	Atomic composition:	C₁₅₆₉H₂₄₁₇N₄₁₉O₄₈₃S₁₃
	Molecular mass [Da]:	35294.7
	Theoretical pI:	5.90
	Extinction coefficient at 280 nm [M^-1 cm^-1]:	47330 / 47455 (all Cys red/ox)

Plot for hydrophobicity, charge, predicted secondary structure, solvent accessability, transmembrane helices and disulfid bridges

Codon usage

	Organism:	E. coli	B. subtilis	S. cerevisiae	A. thaliana	P. patens	Mammals
	Codon quality (CAI):	good (0.80)	good (0.72)	good (0.67)	good (0.75)	good (0.79)	good (0.71)

Alignments (obtained from PredictProtein.org)
There were no alignments for this protein in the data base. The BLAST search was initialized and should be ready in a few hours.

Predictions (obtained from PredictProtein.org)

There were no predictions for this protein in the data base. The prediction was initialized and should be ready in a few hours.

The BioBrick-AutoAnnotator was created by TU-Munich 2013 iGEM team. For more information please see the documentation.
If you have any questions, comments or suggestions, please leave us a comment.

References

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 *edited to show relevant bands
 </p>''' ]]
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-<span class='h3bb'>Sequence and Features</span>
+===Sequence and Features===
 <partinfo>BBa_K2842680 SequenceAndFeatures</partinfo>
 <partinfo>BBa_K2842680 parameters</partinfo>