panS-loxP-MCS
This shuttle vector was modified based on the shuttle vector BBa_K3228089 designed by iGEM Team Marburg 2019. It contains the ori ColE1 for E.coli and the minimal replication region of the pANS plasmid for S.elongatus. The minimal replication region of the pANS plasmid has been codon optimized for S.elongatus UTEX 2973 (see BBa_K3776007). The kanamycin resistance cassette has been codon optimized for S.elongatus UTEX 2973 (see see BBa_K3776008). ColE1 can be removed with Cre recombinases. This allows users to first transform this shuttle vector into E.coli cells for plasmid miniprep then remove the ColE1 ori before transformation into S.elongatus cells. ColE1 has no function in S.elongatus, so removing it reduces energy used for replication of the shuttle vector in S.elongatus. The multiple cloning site (MCS) allows flexibility for cloning.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NotI site found at 33
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 206
Illegal BamHI site found at 6
Illegal XhoI site found at 5663 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 2378
Illegal AgeI site found at 4103 - 1000COMPATIBLE WITH RFC[1000]