BBa_K259004
1
BioScaffol
BioScaffold Linker - Removes Stop Codons & scars & replaces with a Gly-Ser linker
2009-07-31T11:00:00Z
2015-05-08T01:11:42Z
This part is completely designed ''de novo''. This sequence does not occur naturally.
This part is part of a family of Bioscaffold - Linkers.
This part will help users interested in protein-fusion. The usage of the part will allow in frame assembly of 2(+) proteins by:
*Removing the stop codons from the upstream protein.
*Removing the scar created by the Assembly Standard 10 (RFC10) construction of 2(+) protein coding sequences.
*Maintaining the start codon of the downstream protein.
*Adding a flexible (Gly-Ser-Gly) linker between the two protein coding sequences.
*Acting as a translational buffer if ligation is incorrect.
===Assembly Instructions===
*Create an Assembly Standard 10 fusion as below ;
**Upstream Protein Part - Bioscaffold Linker - Downstream Protein Part**
[[Image:BCCS_Assembly_of_Scaffold.jpg|thumb|500px|center|The Product of Assembly between 2 proteins and the Bioscaffold Linker. ]]
*Use the BioScaffold Specific Enzymes
(i)Restrict with BpuEI to remove upstream scar (created between Upstream Protein - Bioscaffold) and stop codons from upstream protein part.<br/>
(ii)Ligate and you will get the intermediate product;
**Upstream Protein (w/out Stop codons) - Tyr - BioScaffold Linker - Downstream Protein**
*Restrict with CspCI to remove downstream scar.
*Ligate to get the final product;
**Upstream Protein - Tyr-Gly-Ser-Gly- Downstream Protein**
[[Image:BCCS_Bioscaffold_Reaction_Explanation_CspCI.jpg|thumb|500px|center|The Reaction and the product. ]]
false
false
_357_
0
5318
9
It's complicated
false
This part was designed taking into consideration the requirements by Assembly Standard 10 (RFC10).
false
Petros Mina
annotation2014601
1
Met Involved in Restriction/Ligation
range2014601
1
53
55
annotation2014605
1
CspCI (3' Recognition Seq)
range2014605
1
74
77
annotation2014597
1
BpuEI
range2014597
1
3
8
annotation2014603
1
CspCI (5' Recognition Seq)
range2014603
1
66
68
annotation2014602
1
Translational Buffer
range2014602
1
56
65
annotation2014604
1
Translational Buffer
range2014604
1
70
72
annotation2014598
1
BpuEI
range2014598
1
21
26
annotation2014599
1
Tyr Involved in Restriction/Ligation
range2014599
1
41
43
annotation2014600
1
Gly-Ser-Gly Linker
range2014600
1
44
52
BBa_K259004_sequence
1
atctcaagagtggcagcggtcttgagtggcagcggcggtatacggcagcggtatgtaagtagtaacaagtagcgtggggca
igem2sbol
1
iGEM to SBOL conversion
Conversion of the iGEM parts registry to SBOL2.1
James Alastair McLaughlin
Chris J. Myers
2017-03-06T15:00:00.000Z