Difference between revisions of "Part:BBa K1968001:Design"

 
 
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===Design Notes===
 
===Design Notes===
−
Nucleotide X at position X was mutated to a X in order to remove a BsaI site, which would have interfered with later assemblies.  
+
Nucleotide G at position 109 was mutated to an A in order to remove a BbsI site, which would have interfered with later MoClo assemblies.
 +
This part contains two MoClo fusion sites, A at the 5' end and C at the 3' end. The reason why the C fusion site rather than the B site is included is because this promoter does not need an RBS and can be cloned directly in front of a reporter gene, which is usually flanked by C and D fusion sites.
 +
A = GGAG; B = TACT; C = AATG; D = AGGT; E = GCTT; F = CGCT; G = TGCC; H = ACTA.
  
  
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===Source===
 
===Source===
  
−
The part was ordered as gblock through IDT and was subsequently cloned into PhytoBricks Universal Acceptor plasmids (BBa_P10500).  
+
The sequence of this part was outlined in (1). This part was ordered as a gBlock Gene Fragment from IDT and was assembled into the PhytoBricks Universal Acceptor (BBa_P10500). The sequence of this part was confirmed by Sanger Sequencing.  
  
 
===References===
 
===References===
 +
(1) Zhou, J., Zhang, H., Meng, H., Zhu, Y., Bao, G., Zhang, Y., . . . Ma, Y. (2014). Discovery of a super-strong promoter enables efficient production of heterologous proteins in cyanobacteria. Sci Rep, 4, 4500. doi: 10.1038/srep04500

Latest revision as of 18:41, 11 October 2016


Synechocystis Pcpc560 Phytobrick promoter


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
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 2


Design Notes

Nucleotide G at position 109 was mutated to an A in order to remove a BbsI site, which would have interfered with later MoClo assemblies. This part contains two MoClo fusion sites, A at the 5' end and C at the 3' end. The reason why the C fusion site rather than the B site is included is because this promoter does not need an RBS and can be cloned directly in front of a reporter gene, which is usually flanked by C and D fusion sites. A = GGAG; B = TACT; C = AATG; D = AGGT; E = GCTT; F = CGCT; G = TGCC; H = ACTA.


Source

The sequence of this part was outlined in (1). This part was ordered as a gBlock Gene Fragment from IDT and was assembled into the PhytoBricks Universal Acceptor (BBa_P10500). The sequence of this part was confirmed by Sanger Sequencing.

References

(1) Zhou, J., Zhang, H., Meng, H., Zhu, Y., Bao, G., Zhang, Y., . . . Ma, Y. (2014). Discovery of a super-strong promoter enables efficient production of heterologous proteins in cyanobacteria. Sci Rep, 4, 4500. doi: 10.1038/srep04500