Difference between revisions of "Part:BBa K2350012:Design"
(→‎Design Notes)
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1. All Pst1 cutting sites in PrbcL were site-directed mutated.
 
1. All Pst1 cutting sites in PrbcL were site-directed mutated.
  
−
2. Fusion PCR primers:
+
 
 +
2. Fusion PCR primers for PrbcL and IndC:
  
 
Forward:CTCTAGGGAGAGACGACATGttagaaaataatattacaca
 
Forward:CTCTAGGGAGAGACGACATGttagaaaataatattacaca
  
 
Rrverse:tgtgtaatattattttctaaCATGTCGTCTCTCCCTAGAG
 
Rrverse:tgtgtaatattattttctaaCATGTCGTCTCTCCCTAGAG
−
 
−
 
  
 
===Source===
 
===Source===

Revision as of 14:22, 21 October 2017


PrbcL-IndC


We fuse IndC with the intrinsic promoter of Rubisco large subunit (PrbcL), which is proved to be functional in cyanobacteria, and cloned into our pigment vector to see whether Indigoidine would express in S. elongatus PCC 7942.



Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1641
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 2904


Design Notes

1. All Pst1 cutting sites in PrbcL were site-directed mutated.


2. Fusion PCR primers for PrbcL and IndC:

Forward:CTCTAGGGAGAGACGACATGttagaaaataatattacaca

Rrverse:tgtgtaatattattttctaaCATGTCGTCTCTCCCTAGAG

Source

IndC is from iGEM distribution kit BBa_K1152008, and PrbcL is from Synechoccocus elongatus PCC7942 genomic DNA.

References

1. Pei-Hong Chen, Hsien-Lin Liu, Yin-Ju Chen, Yi-Hsiang Cheng, Wei-Ling Lin, Chien-Hung Yeh and Chuan-Hsiung Chang (2012). Enhancing CO2 bio-mitigation by genetic engineering of cyanobacteria

2. https://parts.igem.org/wiki/index.php?title=Part:BBa_K1152008