Difference between revisions of "Part:BBa K259002"

Revision as of 17:49, 1 August 2009

BioScaffold Linker - Removes Stop Codons & scars & replaces with a Gly-Ser linker

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**

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.
(ii)Ligate and you will get the intermediate product;

Upstream Protein (w/out Stop codons) - Tyr - BioScaffold Linker - Downstream Protein

Restrict with BseRI to remove downstream scar.
Ligate to get the final product;

  **Upstream Protein - Tyr-Gly-Ser-Gly- Downstream Protein**

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
COMPATIBLE WITH RFC[1000]

@@ Line 1: / Line 1: @@
 __NOTOC__
 <partinfo>BBa_K259002 short</partinfo>
@@ Line 14: / Line 13: @@
 *Acting as a translational buffer if ligation is incorrect.
-===Assembly Instructions====
+===Assembly Instructions===
+*Create an Assembly Standard 10 fusion as below ;
-*Create an Assembly Standard 10 fusion giving ;
+  **Upstream Protein Part - Bioscaffold Linker - Downstream Protein Part**
-**Upstream Protein Part - Bioscaffold Linker - Downstream Protein Part
 *Use the BioScaffold Specific Enzymes
-**Restrict with BpuEI to remove upstream scar (created between Upstream Protein - Bioscaffold) and stop codons from upstream protein part.
+(i)Restrict with BpuEI to remove upstream scar (created between Upstream Protein - Bioscaffold) and stop codons from upstream protein part.<br/>
-**Ligate and you will get the intermediate product;
+(ii)Ligate and you will get the intermediate product;
 Upstream Protein (w/out Stop codons) - Tyr - BioScaffold Linker - Downstream Protein
-**Restrict with BseRI to remove downstream scar.
+*Restrict with BseRI to remove downstream scar.
-**Ligate to get the final product;
+*Ligate to get the final product;
-Upstream Protein - Tyr-Gly-Ser-Gly- Downstream Protein
+   **Upstream Protein - Tyr-Gly-Ser-Gly- Downstream Protein**