Part:BBa_M10041:Design

{a~INP˃} | C-terminal displaying Ice Nucleation Protein

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 650
  Illegal NgoMIV site found at 2162
  Illegal NgoMIV site found at 2306
  Illegal NgoMIV site found at 2570
  Illegal NgoMIV site found at 2714
  Illegal NgoMIV site found at 2762
  Illegal NgoMIV site found at 2882
  Illegal AgeI site found at 421
  Illegal AgeI site found at 1826
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 596
  Illegal BsaI.rc site found at 3342
  Illegal SapI.rc site found at 242
  

Design Notes

Part has included 4 upstream base pairs for preferential encoding the rbs/cds junction, such that the BglBrick scar (GGATCT) will end up superimposed over the Shine-Delgarno sequence. These 4 upstream base pairs come from the 5'UTR (untranslated region) just upstream of the start codon.

Source

Made by PCR based assembly with [http://openwetware.org/images/9/98/PBca1256-Bca1346.seq pBca1256-Bca1346] as the template DNA

Construction of Ice Nucleation Protein

PCR Odd003F/BBa_G00101 on pBca1256-Bca1346		(3722 bp, EcoRI/BamHI)
Sub into pBca9495CA-Bca1144#5				(EcoRI/BamHI, 3039+910 bp, L)
Product is pBca9495CA-M10041				{a~INP>} 
---------------------------
Odd003F	Forward oligo for INP			ccaaaGAATTCatgAGATCTtgtaATGaatctcgacaaggcg
BBa_G00101 	Reverse sequencing of pSB1A* plasmids	attaccgcctttgagtgagc

References

PMID: 14705004
Li, L., D.G. Kang, and H.J. Cha, Functional Display of Foreign Protein on Surface of Escherichia coli Using N-Terminal Domain of Ice Nucleation Protein. Biotechnology and Bioengineering, 2004. 85(2): p. 214-221.