Difference between revisions of "Part:BBa K2010007:Design"

 
 
Line 7: Line 7:
  
 
===Design Notes===
 
===Design Notes===
−
PETase + pelB + sfGFP
+
This sequence was optimized for E. coli K12 using the Codon Optimization Tool from the IDT website.
  
 +
The T7 promoter was used in order to have strong, inducible transcription, in conjunction with NEB’s T7 Express lysY/Iq Competent E. coli.
 +
 +
RBS 34 was used for standard efficiency.
 +
 +
The GGS linker was used to prevent the His-tag from interfering with the folding of PETase.
 +
 +
The His-tag was included for His-tag purification.
 +
 +
sfGFP was included as a reporter protein.
 +
 +
The pelB tag was included for excretion of PETase.
  
  
 
===Source===
 
===Source===
  
−
PETase + pelB + sfGFP
+
T7 promoter: BBa_I712074
 +
 
 +
RBS: BBa_B0034
 +
 
 +
PETase, originally named ISF6_4831, comes from Ideonella sakaiensis, identified in "A bacterium that degrades and assimilates poly(ethylene terephthalate)" (Yoshida et al. 2016).
 +
 
  
 
===References===
 
===References===

Latest revision as of 07:29, 8 October 2016


pelB + sfGFP + PETase (PET-degrading enzyme, origin I. sakaiensis)


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 118
    Illegal NgoMIV site found at 815
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 1233


Design Notes

This sequence was optimized for E. coli K12 using the Codon Optimization Tool from the IDT website.

The T7 promoter was used in order to have strong, inducible transcription, in conjunction with NEB’s T7 Express lysY/Iq Competent E. coli.

RBS 34 was used for standard efficiency.

The GGS linker was used to prevent the His-tag from interfering with the folding of PETase.

The His-tag was included for His-tag purification.

sfGFP was included as a reporter protein.

The pelB tag was included for excretion of PETase.


Source

T7 promoter: BBa_I712074

RBS: BBa_B0034

PETase, originally named ISF6_4831, comes from Ideonella sakaiensis, identified in "A bacterium that degrades and assimilates poly(ethylene terephthalate)" (Yoshida et al. 2016).


References