Difference between revisions of "Part:BBa K249005:Design"

 
(→‎Design Notes)
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<partinfo>BBa_K249005 short</partinfo>
 
<partinfo>BBa_K249005 short</partinfo>
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===Design Notes===
 
===Design Notes===
−
Standard
+
BioFusion (aka Silver Lab) prefix and Standard Biobrick Suffix
 +
 
 +
This part wsas designed as a plasmid backbone to allow for easy attachment of a C-terminal Arginine TAg to a gene of choice for localization into the lumazine Synthase microcompartment.
 +
 
 +
This part was shown to work vis the sequencing information that we received.
 +
 
 +
pSB - C-terminus
 +
>lcl|20071
 +
 
 +
Length=30
 +
Score = 60.0 bits (30), Expect = 1e-15
 +
Identities = 30/30 (100%), Gaps = 0/30 (0%)
 +
Strand=Plus/Plus
 +
 
 +
Query 12 (the plasmid sent in)
 +
CGCCGCCGCCGCCGCCGCCGCCGCCGCTAA 41
 +
 
 +
Sbjct 1 (the sequence expected)
 +
CGCCGCCGCCGCCGCCGCCGCCGCCGCTAA 30
 +
 
 +
We fused a YFP gene with a C-terminal BioFusion standar suffix to this gene
 +
 
 +
C-terminal EYFP-dT
 +
Score = 19.9 bits (10), Expect = 0.80
 +
Identities = 10/10 (100%), Gaps = 0/10 (0%)
 +
 
 +
Strand=Plus/Plus
 +
 
 +
Query 659 (the plasmid sent in)
 +
GAAGTTCATC 668
 +
 
 +
Sbjct 135 (the sequence expected)
 +
GAAGTTCATC 144
 +
 
 +
 
 +
Query 714 ACAGCCACA 722
 +
 
 +
Sbjct 440 ACAGCCACA 448
  
  
 +
The part was sequenced and showed some gaps due to low resolution of the sequencing data. However, it does show that the YFP gene is present, as is the C-terminal Arginine tag. Theses are simply examples of positive hits that came back, although there are many more that are present. In all there were 10 gaps in the sequence, all of which corresponded to N nucleotides in the sequencing information we received.
  
 
===Source===
 
===Source===

Revision as of 14:37, 31 October 2009

C-terminal Arginine Fusion Vector


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
  • 12
    INCOMPATIBLE WITH RFC[12]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
  • 21
    INCOMPATIBLE WITH RFC[21]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
  • 23
    INCOMPATIBLE WITH RFC[23]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
  • 25
    INCOMPATIBLE WITH RFC[25]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.


Design Notes

BioFusion (aka Silver Lab) prefix and Standard Biobrick Suffix

This part wsas designed as a plasmid backbone to allow for easy attachment of a C-terminal Arginine TAg to a gene of choice for localization into the lumazine Synthase microcompartment.

This part was shown to work vis the sequencing information that we received.

pSB - C-terminus >lcl|20071

Length=30 Score = 60.0 bits (30), Expect = 1e-15 Identities = 30/30 (100%), Gaps = 0/30 (0%) Strand=Plus/Plus

Query 12 (the plasmid sent in) CGCCGCCGCCGCCGCCGCCGCCGCCGCTAA 41

Sbjct 1 (the sequence expected) CGCCGCCGCCGCCGCCGCCGCCGCCGCTAA 30

We fused a YFP gene with a C-terminal BioFusion standar suffix to this gene

C-terminal EYFP-dT Score = 19.9 bits (10), Expect = 0.80 Identities = 10/10 (100%), Gaps = 0/10 (0%)

Strand=Plus/Plus

Query 659 (the plasmid sent in) GAAGTTCATC 668

Sbjct 135 (the sequence expected) GAAGTTCATC 144


Query 714 ACAGCCACA 722

Sbjct 440 ACAGCCACA 448


The part was sequenced and showed some gaps due to low resolution of the sequencing data. However, it does show that the YFP gene is present, as is the C-terminal Arginine tag. Theses are simply examples of positive hits that came back, although there are many more that are present. In all there were 10 gaps in the sequence, all of which corresponded to N nucleotides in the sequencing information we received.

Source

Synthesized DNA

References