Part:BBa_K243035
pJS419
The pJS419 vector is used as an expression vector. It has a weak Shine-Dalgano sequence that enables a lower expression level of the target protein compared to the same vector backbone with a strong one. This can be useful for proteins with a complex stucture or even slightly toxic proteins.
Usage and Biology
The vector is used for expression of complex molecules in order not to overburden the expressing host cell. Also, it is used for proteins that tend to aggregate as well as for the expression of slightly toxic proteins. We used this vector containing a chloramphenicol resistance for the expression of the His-Dig-Split-Fok_a construct (BBa_K243036). Simultaneous expression of the other heterodimeric Fok construct His-FluA-Split-Fok_i (BBa_K243010) took place in a different vector pEx with an ampicillin resistance gene. This enables a positive selection of the bacteria strain XL1 blue cotransformed with both plasmids.
The left picture shows a map of the vector with an ampicillin resistance gene inserted. Due to the limited distance of the start codon to the included ribosome binding site, some restriction sites of the assembly standard 25 have been avoided during the design step.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal suffix found in sequence at 2266
Illegal XbaI site found at 1382 - 12INCOMPATIBLE WITH RFC[12]Illegal SpeI site found at 2267
Illegal PstI site found at 2281
Illegal NotI site found at 2274 - 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 1494
Illegal BamHI site found at 2365 - 23INCOMPATIBLE WITH RFC[23]Illegal suffix found in sequence at 2267
Illegal XbaI site found at 1382 - 25INCOMPATIBLE WITH RFC[25]Illegal suffix found in sequence at 2257
Illegal XbaI site found at 1382
Illegal NgoMIV site found at 1390
Illegal NgoMIV site found at 2498
Illegal AgeI site found at 3832 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 2114
| None |