Part:BBa_K863207:Design
tvel5 integrated in shuttle vector pECPP11JS (BBa_K863204) for integration in yeast
- 10INCOMPATIBLE WITH RFC[10]Illegal XbaI site found at 5558
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 4853
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 3
Illegal BglII site found at 1931
Illegal BglII site found at 2363
Illegal BglII site found at 3004
Illegal BamHI site found at 3361
Illegal XhoI site found at 1198
Illegal XhoI site found at 1963
Illegal XhoI site found at 2596 - 23INCOMPATIBLE WITH RFC[23]Illegal XbaI site found at 5558
- 25INCOMPATIBLE WITH RFC[25]Illegal XbaI site found at 5558
Illegal NgoMIV site found at 2449
Illegal NgoMIV site found at 2602
Illegal AgeI site found at 1330 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 5714
Illegal BsaI.rc site found at 1885
Illegal BsaI.rc site found at 3513
Illegal SapI.rc site found at 1395
Design Notes
Primers for isolation of tvel5 BBa_K863030 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'-acgtcacctgcgtgtagctggtatcggtcctgtcgccg-3'
rev: 5'-acgtcacctgcgtgtcaagTTACTGGTCGCTCGGGTCGC-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_K863030 integrated in the shuttle vector pECPP11JS (https://parts.igem.org/Part:BBa_K863204).