LOCUS       Exported                8152 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Mutagenesis vector pCAM45, complete sequence.
ACCESSION   AY680862
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8152)
  AUTHORS   May JP, Walker CA, Maskell DJ, Slater JD.
  TITLE     Development of an in vivo Himar1 transposon mutagenesis system for 
            use in Streptococcus equi subsp. equi
  JOURNAL   FEMS Microbiol. Lett. 238 (2), 401-409 (2004)
  PUBMED    15358426
REFERENCE   2  (bases 1 to 8152)
  AUTHORS   May JP, Maskell DJ, Slater JD.
  TITLE     Direct Submission
  JOURNAL   Submitted (08-JUL-2004) Clinical Veterinary Medicine, University of 
            Cambridge, Madingley Road, Cambridge CB3 0ES, UK
REFERENCE   3  (bases 1 to 8152)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8152
                     /organism="Mutagenesis vector pCAM45"
                     /mol_type="other DNA"
                     /db_xref="taxon:286693"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   622..650
     CDS             864..1601
                     /codon_start=1
                     /gene="ermAM"
                     /product="ErmAM"
                     /label=ermAM
                     /note="erythromycin resistance determinant"
                     /protein_id="AAT79488.1"
                     /translation="MNKNIKYSQNFLTSEKVLNQIIKQLNLKETDTVYEIGTGKGHLTT
                     KLAKISKQVTSIELDSHLFNLSSEKLKLNTRVTLIHQDILQFQFPNKQRYKIVGSIPYH
                     LSTQIIKKVVFESHASDIYLIVEEGFYKRTLDIHRTLGLLLHTQVSIQQLLKLPAECFH
                     PKPKVNSVLIKLTRHTTDVPDKYWKLYTYFVSKWVNREYRQLFTKNQFHQAMKHAKVNN
                     LSTVTYEQVLSIFNSYLLFNGRK"
     gene            864..1601
                     /gene="ermAM"
                     /label=ermAM
     rep_origin      complement(1675..1993)
                     /direction=LEFT
                     /label=R6K gamma
                     /note="R6K gamma"
     CDS             complement(2019..2279)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSKRESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFAS
                     WRNSEEARTDRPSQQLRSLFPMMSTFKVLLTGWLISPRSDT"
     repeat_region   2028..2056
     primer_bind     2216..2232
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2275..2291)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2299..2315
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(2323..2353)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2368..2389
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(2677..3265)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3764..3775)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             4036..5082
                     /codon_start=1
                     /product="mariner transposase C9 mutant"
                     /label=mariner transposase C9 mutant
                     /protein_id="AAT79489.1"
                     /translation="MEKKEFRVLIKYCFLKGKNTVEAKTWLDNEFPDSAPGKSTIIDWY
                     AKFKRGEMSTEDGERSGRPKEVVTDENIKKIHKMILNDRKMKLIEIAEALKISKERVGH
                     IIHQYLDMRKLCAKWVPRELTFDQKQRRVDDSKRCLQLLTRNTPEFFRRYVTMDETWLH
                     HYTPESNRQSAEWTATGEPSPKRGKTQKSAGKVMASVFWDAHGIIFIDYLEKGKTINSD
                     YYMALLERLKVEIAAKRPHMKKKKVLFHQDNAPCHKSLRTMAKIHELGFELLPHPPYSP
                     DLAPSDFFLFSDIKRMLAGKKFGCNEEVIAETEAYFEAKPKEYYQNGIKKLEGRYNRCI
                     ALEGNYVE"
     CDS             5443..5604
                     /codon_start=1
                     /gene="repC"
                     /product="RepC"
                     /label=repC
                     /protein_id="AAT79490.1"
                     /translation="MVISESKKRVMISLTKEQDKKLTDMAKQKGFSKSAVAALAIEEYA
                     RKESEQKK"
     gene            5443..5604
                     /gene="repC"
                     /label=repC
     CDS             complement(6773..7567)
                     /codon_start=1
                     /gene="aphA3"
                     /product="aminoglycoside phosphotransferase A3"
                     /label=aphA3
                     /protein_id="AAT79491.1"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     gene            complement(6773..7567)
                     /gene="aphA3"
                     /label=aphA3