Plasmid
Part:BBa_K3731004:Design
Designed by: Hao Yin Group: iGEM21_Nanjing-China (2021-09-29)
EPVM-PBBR1MCS-2
Assembly Compatibility:
- 10INCOMPATIBLE WITH RFC[10]Illegal prefix found in sequence at 5166
Illegal suffix found in sequence at 1 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 5166
Illegal SpeI site found at 2
Illegal PstI site found at 16
Illegal NotI site found at 9
Illegal NotI site found at 5172 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 5166
- 23INCOMPATIBLE WITH RFC[23]Illegal prefix found in sequence at 5166
Illegal suffix found in sequence at 2 - 25INCOMPATIBLE WITH RFC[25]Illegal prefix found in sequence at 5166
Illegal XbaI site found at 5181
Illegal SpeI site found at 2
Illegal PstI site found at 16 - 1000COMPATIBLE WITH RFC[1000]
Design Notes
PCR-amplified CFppk1-vgb-mazE was digested with restriction enzymes Kpn I and Hind III, after which it was insert into corresponding multiple cloning sties of the broad-host-range expressing vector pBBR1MCS2.
Source
For the construction of EPVM-PBBR1MCS-2, genomic DNA of E.Coli BL21 was used as the template to PCR-amplify ppk1, vgb and mazE with primers.