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

(→‎Design Notes)
(→‎Design Notes)
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This part was build by a Gibson assembly. 5' overlap parts were created by PCR with the following primers.
 
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 [https://parts.igem.org/Part:BBa_K863200 K863200] was created with the primers:
+
The 5' UTR of alcohol oxidase 1 gene (AOX1) of Pichia pastoris was created with the primers:
  
 
FW: 5'-CGCTAAGGATGATTTCTGGAATTCGCGGCCGCTTCTAGAGAGATCTAACATCCAAAGACG-3'
 
FW: 5'-CGCTAAGGATGATTTCTGGAATTCGCGGCCGCTTCTAGAGAGATCTAACATCCAAAGACG-3'
Line 15: Line 15:
  
  
−
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 was created with the primers:
  
 
FW: 5'-TTATTTAGAGATTTTAACTTACATTTAGATTCGATAGATCCGAGTATCTATGATTGGAAG-3'
 
FW: 5'-TTATTTAGAGATTTTAACTTACATTTAGATTCGATAGATCCGAGTATCTATGATTGGAAG-3'
Line 22: Line 22:
  
  
−
The his4 gene from Pichia pastoris [https://parts.igem.org/Part:BBa_K863202 K863202] was created with the primers:
+
The his4 gene from Pichia pastoris was created with the primers:
  
 
FW: 5'-GTACAGAAGATTAAGTGAGAAGTTCGTTTGTGCAAGCTTAAGATCTCCTGATGACTGACTC-3'
 
FW: 5'-GTACAGAAGATTAAGTGAGAAGTTCGTTTGTGCAAGCTTAAGATCTCCTGATGACTGACTC-3'
Line 29: Line 29:
  
  
−
The mating factor alpha 1 with Kozak sequence [https://parts.igem.org/Part:BBa_K863206 K863206] was created with the primers:
+
The mating factor alpha 1 with Kozak sequence was created with the primers:
  
 
FW: 5'-AGAAGATCAAAAAACAACTAATTATTCGAAACGCCCGCCGCCACCATGAGATTTCCTTCAATTTTTAC-3'
 
FW: 5'-AGAAGATCAAAAAACAACTAATTATTCGAAACGCCCGCCGCCACCATGAGATTTCCTTCAATTTTTAC-3'
Line 36: Line 36:
  
  
−
The AOX1 Terminator from Pichia pastoris [https://parts.igem.org/Part:BBa_K863203 K863203] was created with the primers:
+
The AOX1 Terminator from Pichia pastoris was created with the primers:
  
 
FW: 5'-GTATCTCTCGAGAAAAGAGAGGCTGAAGCTACACGCAGGTGGTATGTATCACCTGCGTGTCTTGCTAGATTCTAATCAAG-3'
 
FW: 5'-GTATCTCTCGAGAAAAGAGAGGCTGAAGCTACACGCAGGTGGTATGTATCACCTGCGTGTCTTGCTAGATTCTAATCAAG-3'
Line 44: Line 44:
  
  
−
 
+
All components were created as BioBricks for the community:
−
 
+
* 5' UTR of alcohol oxidase 1 gene (AOX1) of Pichia pastoris [https://parts.igem.org/Part:BBa_K863200 K863200]
−
 
+
* 3' UTR of alcohol oxidase 1 gene (AOX1) of Pichia pastoris [https://parts.igem.org/Part:BBa_K863201 K863201]
−
 
+
* his4 gene from Pichia pastoris [https://parts.igem.org/Part:BBa_K863202 K863202]
−
 
+
* mating factor alpha 1 with Kozak sequence [https://parts.igem.org/Part:BBa_K863206 K863206]
−
 
+
* AOX1 Terminator from Pichia pastoris [https://parts.igem.org/Part:BBa_K863203 K863203]
−
All components were created as Parts for the iGEM community:
+
  
 
===Source===
 
===Source===

Revision as of 03:28, 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 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 was created with the primers:

FW: 5'-TTATTTAGAGATTTTAACTTACATTTAGATTCGATAGATCCGAGTATCTATGATTGGAAG-3'

RV: 5'-AAAACAAGATAGTGCCCCTC-3'


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

FW: 5'-GTACAGAAGATTAAGTGAGAAGTTCGTTTGTGCAAGCTTAAGATCTCCTGATGACTGACTC-3'

RV: 5'-CTCGGATCTATCGAATCTAAATGTAAG-3'


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

FW: 5'-AGAAGATCAAAAAACAACTAATTATTCGAAACGCCCGCCGCCACCATGAGATTTCCTTCAATTTTTAC-3'

RV: 5'-AGCTTCAGCCTCTCTTTTCT-3'


The AOX1 Terminator from Pichia pastoris was created with the primers:

FW: 5'-GTATCTCTCGAGAAAAGAGAGGCTGAAGCTACACGCAGGTGGTATGTATCACCTGCGTGTCTTGCTAGATTCTAATCAAG-3'

RV: 5'-TAAGCTTGCACAAACGAACT-3'


All components were created as BioBricks for the community:

  • 5' UTR of alcohol oxidase 1 gene (AOX1) of Pichia pastoris K863200
  • 3' UTR of alcohol oxidase 1 gene (AOX1) of Pichia pastoris K863201
  • his4 gene from Pichia pastoris K863202
  • mating factor alpha 1 with Kozak sequence K863206
  • AOX1 Terminator from Pichia pastoris K863203

Source

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

References