Part:BBa_K863208:Design
tvel35 integrated in shuttle vector pECPP11JS (BBa_K863204) for integration in yeast
- 10INCOMPATIBLE WITH RFC[10]Illegal XbaI site found at 5552
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 1621
Illegal NheI site found at 1969
Illegal NheI site found at 4847 - 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 3
Illegal BglII site found at 2998
Illegal BamHI site found at 1994
Illegal BamHI site found at 3355
Illegal XhoI site found at 1198 - 23INCOMPATIBLE WITH RFC[23]Illegal XbaI site found at 5552
- 25INCOMPATIBLE WITH RFC[25]Illegal XbaI site found at 5552
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 5708
Illegal BsaI.rc site found at 2539
Illegal BsaI.rc site found at 3507
Design Notes
Primers for isolation of tvel5 BBa_K863034 were designed with AarI overhanging ends before and after the gene sequence. Additionally the gene sequence in the primers is without ATG and starts after signal sequence, which is normally included in the gene sequence.
fwd: 5'-acgtcacctgcgtgtagctgccatagggcctgtggcgg-3'
rev: 5'-acgtcacctgcgtgtcaagTCAGAGGTCGCTGGGGTCAA-3'
The PCR product was digested with [http://www.fermentas.de/product_info.php?info=p166&language=de AarI] and ligated in BBa_K863204, which has to be digested with AarI, too. For further information about the design of BBa_K863204 look here.
Source
This part is composed of BBa_K863034 integrated in the shuttle vector pECPP11JS (BBa_K863204).