LOCUS       Exported                6100 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Integration vector pMTL-JH20, complete sequence.
ACCESSION   HQ875761
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6100)
  AUTHORS   Heap JT, Ehsaan M, Cooksley CM, Ng YK, Cartman ST, Winzer K, Minton 
            NP.
  TITLE     Integration of DNA into bacterial chromosomes from plasmids without 
            a counter-selection marker
  JOURNAL   Nucleic Acids Res. 40 (8), E59 (2012)
  PUBMED    22259038
REFERENCE   2  (bases 1 to 6100)
  AUTHORS   Heap JT, Ehsaan M, Cooksley CM, Cartman ST, Minton NP.
  TITLE     Direct Submission
  JOURNAL   Submitted (11-JAN-2011) School of Molecular Medical Sciences, The 
            University of Nottingham, Centre for Biomolecular Sciences, 
            University Park, Nottingham, Nottinghamshire NG7 2RD, United Kingdom
REFERENCE   3  (bases 1 to 6100)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6100
                     /organism="Integration vector pMTL-JH20"
                     /mol_type="other DNA"
                     /db_xref="taxon:1128732"
     gene            9..546
                     /gene="pyrE fragment"
                     /label=pyrE fragment
     misc_feature    9..308
                     /gene="pyrE fragment"
                     /note="mediates homologous recombination with the 
                     chromosome of Clostridium difficile 630"
     misc_feature    564..827
                     /label=multiple cloning site
                     /note="multiple cloning site"
     CDS             564..827
                     /codon_start=1
                     /gene="lacZ"
                     /product="beta-galactosidase alpha subunit"
                     /label=lacZ
                     /protein_id="AFA34969.1"
                     /translation="MTMITNSSSVPGDPLESTSRVHGDLEACRHASLALAVVLQRRDWE
                     NPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWR"
     gene            564..827
                     /gene="lacZ"
                     /label=lacZ
     primer_bind     complement(665..682)
                     /label=M13F
                     /note="M13F"
     primer_bind     complement(665..681)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    829..2028
                     /note="mediates homologous recombination with the 
                     chromosome of Clostridium difficile 630"
     CDS             1040..1966
                     /codon_start=1
                     /gene="CD0188"
                     /product="putative ketopantoate reductase"
                     /label=CD0188
                     /protein_id="AFA34970.1"
                     /translation="MKNQLNENNKKVVFFGVGAVGATFAEQFFNSKYDFKILCDNERKK
                     RYLEEGFIINGKRYDFDYVTKDEYKQEADFIIIGLKYNNLKENIKELDGLVGKNTVIMS
                     LLNGVDSEEIIGERFGIEKMVYSYVTNIDAKKINNNIIHTTNGIIVFGNKDNSEDRKTN
                     IITEVFDDVNIEYTLSKDIQRDMWWKYMVNIGVNQTSAILGAPYGVFQSSEHLRELAKS
                     AMREVVAIAQAKDISLTEDDVEHSLHRILEHSKEGRTSMLQDVEAHRLTEVDMFSKNIC
                     KLGKKYNIPTPINQTFFYMIKVIESRF"
     gene            1040..1966
                     /gene="CD0188"
                     /label=CD0188
     CDS             2755..3072
                     /codon_start=1
                     /gene="repH"
                     /product="plasmid replication protein"
                     /label=repH
                     /protein_id="AFA34971.1"
                     /translation="MKRRAYMKTCKNCKEFIKDTEICKIHSLMIHDKTVATYCSKYNES
                     RCLHKGKVQCINCSKMNRYGWCAIKMRCFTEEEQKKERTCIKYYARSFKKAHVKKSKKK
                     K"
     gene            2755..3072
                     /gene="repH"
                     /label=repH
     CDS             3744..4367
                     /codon_start=1
                     /gene="catP"
                     /product="chloramphenicol acetyltransferase"
                     /label=catP
                     /protein_id="AFA34972.1"
                     /translation="MVFEKIDKNSWNRKEYFDHYFASVPCTYSMTVKVDITQIKEKGMK
                     LYPAMLYYIAMIVNRHSEFRTAINQDGELGIYDEMIPSYTIFHNDTETFSSLWTECKSD
                     FKSFLADYESDTQRYGNNHRMEGKPNAPENIFNVSMIPWSTFDGFNLNLQKGYDYLIPI
                     FTMGKYYKEDNKIILPLAIQVHHAVCDGFHICRFVNELQELINS"
     gene            3744..4367
                     /gene="catP"
                     /label=catP
     rep_origin      4544..5132
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      4580..5125
                     /label=ColE1 RNA II
                     /note="ColE1 RNA II"
     oriT            5453..5562
                     /note="incP origin of transfer"
     CDS             5526..5966
                     /codon_start=1
                     /gene="traJ"
                     /product="plasmid conjugative transfer protein"
                     /label=traJ
                     /protein_id="AFA34973.1"
                     /translation="MLNGNPALRGWPATAGVTDEGKRMADETKPTRKGSPPIKVYCLPD
                     ERRAIEEKAAAAGMSLSAYLLAVGQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWL
                     TDDPRTARFGDATILALLAKIEEKQDELGKVMMGVVRPRAEP"
     gene            5526..5966
                     /gene="traJ"
                     /label=traJ
     CDS             5595..5966
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"