Difference between revisions of "Part:BBa K2350012:Design"

 
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__NOTOC__
 
__NOTOC__
 
<partinfo>BBa_K2350012 short</partinfo>
 
<partinfo>BBa_K2350012 short</partinfo>
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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.
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<partinfo>BBa_K2350012 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K2350012 SequenceAndFeatures</partinfo>
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===Design Notes===
 
===Design Notes===
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All Pst1 cutting sites in PrbcL were site-directed mutated.
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1. All Pst1 cutting sites in PrbcL were site-directed mutated.
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2. Fusion PCR primers:
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Forward:CTCTAGGGAGAGACGACATGttagaaaataatattacaca
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Rrverse:tgtgtaatattattttctaaCATGTCGTCTCTCCCTAGAG
  
  
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===References===
 
===References===
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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
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2. https://parts.igem.org/wiki/index.php?title=Part:BBa_K1152008

Revision as of 14:21, 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:

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