Difference between revisions of "Part:BBa K243033"
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| − | This is our main E. coli protein expression vector. | + | This is our main E. coli protein expression vector. To insert DNA fragments, the plasmid possesses the restriction sites XbaI NgoMIV and AgeI_SpeI_NotI_PstI. The IPTG-inducable plasmid vector contains the pBR322 origin of replication and a Shine-Dalgarno-Sequence. |
Latest revision as of 19:36, 23 October 2009
pEX
This is our main E. coli protein expression vector. To insert DNA fragments, the plasmid possesses the restriction sites XbaI NgoMIV and AgeI_SpeI_NotI_PstI. The IPTG-inducable plasmid vector contains the pBR322 origin of replication and a Shine-Dalgarno-Sequence.
Usage and Biology
We used this vector containing an ampicillin resistance gene for the expression of single constructs such as the His-FluA-Split-Fok_i (BBa_K243010)construct. Simultaneous expression of the other Fok construct, His-Dig-Split-Fok_a (BBa_K243036) could be assured using the vector pJS419 since it confers chloramphenicol resistance to the host cell. This enables a positive selection of bacterial strains such as XL1-blue (Stratagene) cotransformed with both plasmids.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal suffix found in sequence at 2734
Illegal XbaI site found at 1523 - 12INCOMPATIBLE WITH RFC[12]Illegal SpeI site found at 2735
Illegal PstI site found at 2749
Illegal NotI site found at 2742 - 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 1808
- 23INCOMPATIBLE WITH RFC[23]Illegal suffix found in sequence at 2735
Illegal XbaI site found at 1523 - 25INCOMPATIBLE WITH RFC[25]Illegal suffix found in sequence at 2725
Illegal XbaI site found at 1523
Illegal NgoMIV site found at 1531 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 3800
Illegal SapI.rc site found at 4799