Difference between revisions of "Part:BBa K863204:Design"

(→‎Design Notes)
(→‎Design Notes)
Line 13: Line 13:
  
 
RV: 5'-GGTGGCGGCGGGCGTTTCGAATAATTAGTT-3'
 
RV: 5'-GGTGGCGGCGGGCGTTTCGAATAATTAGTT-3'
 +
  
 
The 3' UTR of alcohol oxidase 1 gene (AOX1) of Pichia pastoris [https://parts.igem.org/Part:BBa_K863201 K863201] was created with the primers:
 
The 3' UTR of alcohol oxidase 1 gene (AOX1) of Pichia pastoris [https://parts.igem.org/Part:BBa_K863201 K863201] was created with the primers:
  
−
FW: 5'-TCACCGCAATGCTGTTAAGGTTCGTATGGAGAAATAATAACGAGTATCTATGATTGGAAG-3'
+
FW: 5'-TTATTTAGAGATTTTAACTTACATTTAGATTCGATAGATCCGAGTATCTATGATTGGAAG-3'
  
 
RV: 5'-AAAACAAGATAGTGCCCCTC-3'
 
RV: 5'-AAAACAAGATAGTGCCCCTC-3'
  
 +
 +
The his4 gene from Pichia pastoris [https://parts.igem.org/Part:BBa_K863202 K863202] was created with the primers:
 +
 +
FW: 5'-GTACAGAAGATTAAGTGAGAAGTTCGTTTGTGCAAGCTTAAGATCTCCTGATGACTGACTC -3'
 +
 +
RV: 5'-CTCGGATCTATCGAATCTAAATGTAAG -3'
 +
 +
 +
The mating factor alpha 1 with Kozak sequence [https://parts.igem.org/Part:BBa_K863206 K863206] was created with the primers:
 +
 +
FW: 5'-AGAAGATCAAAAAACAACTAATTATTCGAAACGCCCGCCGCCACCATGAGATTTCCTTCAATTTTTAC  -3'
 +
 +
RV: 5'-AGCTTCAGCCTCTCTTTTCT-3'
 +
 +
 +
The mating factor alpha 1 with Kozak sequence [https://parts.igem.org/Part:BBa_K863206 K863206] was created with the primers:
 +
 +
FW: 5'-AGAAGATCAAAAAACAACTAATTATTCGAAACGCCCGCCGCCACCATGAGATTTCCTTCAATTTTTAC  -3'
 +
 +
RV: 5'-AGCTTCAGCCTCTCTTTTCT-3'
  
  

Revision as of 03:20, 27 September 2012

shuttle vector pECPP11JS for recombination of genes of interest in yeast


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 4086
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 3381
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1
    Illegal BglII site found at 1532
    Illegal BamHI site found at 1889
    Illegal XhoI site found at 1196
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 4086
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 4086
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 4242
    Illegal BsaI.rc site found at 2041


Design Notes

This part was build by a Gibson assembly. 5' overlap parts were created by PCR with the following primers.

The 5' UTR of alcohol oxidase 1 gene (AOX1) of Pichia pastoris K863200 was created with the primers:

FW: 5'-CGCTAAGGATGATTTCTGGAATTCGCGGCCGCTTCTAGAGAGATCTAACATCCAAAGACG-3'

RV: 5'-GGTGGCGGCGGGCGTTTCGAATAATTAGTT-3'


The 3' UTR of alcohol oxidase 1 gene (AOX1) of Pichia pastoris K863201 was created with the primers:

FW: 5'-TTATTTAGAGATTTTAACTTACATTTAGATTCGATAGATCCGAGTATCTATGATTGGAAG-3'

RV: 5'-AAAACAAGATAGTGCCCCTC-3'


The his4 gene from Pichia pastoris K863202 was created with the primers:

FW: 5'-GTACAGAAGATTAAGTGAGAAGTTCGTTTGTGCAAGCTTAAGATCTCCTGATGACTGACTC -3'

RV: 5'-CTCGGATCTATCGAATCTAAATGTAAG -3'


The mating factor alpha 1 with Kozak sequence K863206 was created with the primers:

FW: 5'-AGAAGATCAAAAAACAACTAATTATTCGAAACGCCCGCCGCCACCATGAGATTTCCTTCAATTTTTAC -3'

RV: 5'-AGCTTCAGCCTCTCTTTTCT-3'


The mating factor alpha 1 with Kozak sequence K863206 was created with the primers:

FW: 5'-AGAAGATCAAAAAACAACTAATTATTCGAAACGCCCGCCGCCACCATGAGATTTCCTTCAATTTTTAC -3'

RV: 5'-AGCTTCAGCCTCTCTTTTCT-3'





All components were created as Parts for the iGEM community:

Source

This part is made of the BioBricks XYZ by gibson assembly.

References