LOCUS       Exported                6760 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Allelic exchange vector pCM184, complete sequence.
ACCESSION   AY093429
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6760)
  AUTHORS   Marx CJ, Lidstrom ME.
  TITLE     A broad-host-range cre-lox system for antibiotic marker recycling in
            Gram-negative bacteria
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6760)
  AUTHORS   Marx CJ, Lidstrom ME.
  TITLE     Direct Submission
  JOURNAL   Submitted (28-MAR-2002) Chemical Engineering, University of 
            Washington, Box 351750, Seattle, WA 98195-1750, USA
REFERENCE   3  (bases 1 to 6760)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6760
                     /organism="Allelic exchange vector pCM184"
                     /mol_type="other DNA"
                     /db_xref="taxon:192853"
     CDS             complement(1..861)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /note="ampicillin resistance"
                     /protein_id="AAM19718.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(1..861)
                     /gene="bla"
                     /label=bla
     promoter        complement(862..966)
                     /gene="bla"
                     /label=AmpR promoter
     protein_bind    1050..1083
                     /label=loxP binding site
                     /bound_moiety="loxP"
                     /note="Cre recombinase recognition site"
     CDS             1217..2032
                     /codon_start=1
                     /gene="kan"
                     /product="kanamycin resistance"
                     /label=kan
                     /protein_id="AAM19719.1"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     gene            1217..2032
                     /gene="kan"
                     /label=kan
     protein_bind    2343..2376
                     /label=loxP binding site
                     /bound_moiety="loxP"
                     /note="Cre recombinase recognition site"
     oriT            2941..3679
                     /label=IncP origin of transfer
                     /note="IncP origin of transfer"
     CDS             complement(3031..3402)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            3435..3544
                     /note="incP origin of transfer"
     promoter        4181..4209
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             4257..5447
                     /codon_start=1
                     /gene="tet"
                     /product="tetracyline resistance"
                     /label=tet
                     /protein_id="AAM19720.1"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     gene            4257..5447
                     /gene="tet"
                     /label=tet
     rep_origin      complement(5960..6515)
                     /direction=LEFT
                     /note="ColE1 ori; ColE1 origin of replication"
     rep_origin      complement(6002..6590)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"