Difference between revisions of "Part:BBa K863204:Design"
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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 [https://parts.igem.org/Part:BBa_K863202 K863202] was created with the primers: | ||
− | FW: 5'-GTACAGAAGATTAAGTGAGAAGTTCGTTTGTGCAAGCTTAAGATCTCCTGATGACTGACTC -3' | + | FW: 5'-GTACAGAAGATTAAGTGAGAAGTTCGTTTGTGCAAGCTTAAGATCTCCTGATGACTGACTC-3' |
− | RV: 5'-CTCGGATCTATCGAATCTAAATGTAAG -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: | The mating factor alpha 1 with Kozak sequence [https://parts.igem.org/Part:BBa_K863206 K863206] was created with the primers: | ||
− | FW: 5'-AGAAGATCAAAAAACAACTAATTATTCGAAACGCCCGCCGCCACCATGAGATTTCCTTCAATTTTTAC | + | FW: 5'-AGAAGATCAAAAAACAACTAATTATTCGAAACGCCCGCCGCCACCATGAGATTTCCTTCAATTTTTAC-3' |
RV: 5'-AGCTTCAGCCTCTCTTTTCT-3' | RV: 5'-AGCTTCAGCCTCTCTTTTCT-3' | ||
− | The | + | The AOX1 Terminator from Pichia pastoris [https://parts.igem.org/Part:BBa_K863203 K863203] was created with the primers: |
− | FW: 5'- | + | FW: 5'-GTATCTCTCGAGAAAAGAGAGGCTGAAGCTACACGCAGGTGGTATGTATCACCTGCGTGTCTTGCTAGATTCTAATCAAG-3' |
− | RV: 5'- | + | RV: 5'-TAAGCTTGCACAAACGAACT-3' |
Revision as of 03:23, 27 September 2012
shuttle vector pECPP11JS for recombination of genes of interest in yeast
- 10INCOMPATIBLE WITH RFC[10]Illegal XbaI site found at 4086
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 3381
- 21INCOMPATIBLE 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 - 23INCOMPATIBLE WITH RFC[23]Illegal XbaI site found at 4086
- 25INCOMPATIBLE WITH RFC[25]Illegal XbaI site found at 4086
- 1000INCOMPATIBLE 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 AOX1 Terminator from Pichia pastoris K863203 was created with the primers:
FW: 5'-GTATCTCTCGAGAAAAGAGAGGCTGAAGCTACACGCAGGTGGTATGTATCACCTGCGTGTCTTGCTAGATTCTAATCAAG-3'
RV: 5'-TAAGCTTGCACAAACGAACT-3'
All components were created as Parts for the iGEM community:
Source
This part is made of the BioBricks XYZ by gibson assembly.