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__NOTOC__
 
__NOTOC__
 
<partinfo>BBa_K3776009 short</partinfo>
 
<partinfo>BBa_K3776009 short</partinfo>
  
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improved on 2019 Marburg
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This shuttle vector was modified based on the shuttle vector <a href="https://parts.igem.org/Part:BBa_K3228089"> BBa_K3228089</a> 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 <a href="https://parts.igem.org/Part:BBa_K3776007"> BBa_K3776007</a>). The kanamycin resistance cassette has been codon optimized for S.elongatus UTEX 2973 (see see <a href="https://parts.igem.org/Part:BBa_K3776008"> BBa_K3776008</a>). 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.
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<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Latest revision as of 15:38, 17 October 2021


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


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NotI site found at 33
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 206
    Illegal BamHI site found at 6
    Illegal XhoI site found at 5663
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 2378
    Illegal AgeI site found at 4103
  • 1000
    COMPATIBLE WITH RFC[1000]