LOCUS       Exported                5862 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Integration vector pMTL-JH18, complete sequence.
ACCESSION   HQ875759
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5862)
  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 5862)
  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 5862)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5862
                     /organism="Integration vector pMTL-JH18"
                     /mol_type="other DNA"
                     /db_xref="taxon:1128729"
     misc_feature    9..308
                     /gene="pyrE fragment"
                     /note="mediates homologous recombination with the 
                     chromosome of Clostridium difficile 630"
     gene            9..308
                     /gene="pyrE fragment"
                     /label=pyrE fragment
     misc_feature    326..589
                     /label=multiple cloning site
                     /note="multiple cloning site"
     CDS             326..589
                     /codon_start=1
                     /gene="lacZ"
                     /product="beta-galactosidase alpha subunit"
                     /label=lacZ
                     /protein_id="AFA34960.1"
                     /translation="MTMITNSSSVPGDPLESTSRVHGDLEACRHASLALAVVLQRRDWE
                     NPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWR"
     gene            326..589
                     /gene="lacZ"
                     /label=lacZ
     primer_bind     complement(427..444)
                     /label=M13F
                     /note="M13F"
     primer_bind     complement(427..443)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    591..1790
                     /note="mediates homologous recombination with the 
                     chromosome of Clostridium difficile 630"
     CDS             802..1728
                     /codon_start=1
                     /gene="CD0188"
                     /product="putative ketopantoate reductase"
                     /label=CD0188
                     /protein_id="AFA34961.1"
                     /translation="MKNQLNENNKKVVFFGVGAVGATFAEQFFNSKYDFKILCDNERKK
                     RYLEEGFIINGKRYDFDYVTKDEYKQEADFIIIGLKYNNLKENIKELDGLVGKNTVIMS
                     LLNGVDSEEIIGERFGIEKMVYSYVTNIDAKKINNNIIHTTNGIIVFGNKDNSEDRKTN
                     IITEVFDDVNIEYTLSKDIQRDMWWKYMVNIGVNQTSAILGAPYGVFQSSEHLRELAKS
                     AMREVVAIAQAKDISLTEDDVEHSLHRILEHSKEGRTSMLQDVEAHRLTEVDMFSKNIC
                     KLGKKYNIPTPINQTFFYMIKVIESRF"
     gene            802..1728
                     /gene="CD0188"
                     /label=CD0188
     CDS             2517..2834
                     /codon_start=1
                     /gene="repH"
                     /product="plasmid replication protein"
                     /label=repH
                     /protein_id="AFA34962.1"
                     /translation="MKRRAYMKTCKNCKEFIKDTEICKIHSLMIHDKTVATYCSKYNES
                     RCLHKGKVQCINCSKMNRYGWCAIKMRCFTEEEQKKERTCIKYYARSFKKAHVKKSKKK
                     K"
     gene            2517..2834
                     /gene="repH"
                     /label=repH
     CDS             3506..4129
                     /codon_start=1
                     /gene="catP"
                     /product="chloramphenicol acetyltransferase"
                     /label=catP
                     /protein_id="AFA34963.1"
                     /translation="MVFEKIDKNSWNRKEYFDHYFASVPCTYSMTVKVDITQIKEKGMK
                     LYPAMLYYIAMIVNRHSEFRTAINQDGELGIYDEMIPSYTIFHNDTETFSSLWTECKSD
                     FKSFLADYESDTQRYGNNHRMEGKPNAPENIFNVSMIPWSTFDGFNLNLQKGYDYLIPI
                     FTMGKYYKEDNKIILPLAIQVHHAVCDGFHICRFVNELQELINS"
     gene            3506..4129
                     /gene="catP"
                     /label=catP
     rep_origin      4306..4894
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      4342..4887
                     /label=ColE1 RNA II
                     /note="ColE1 RNA II"
     oriT            5215..5324
                     /note="incP origin of transfer"
     CDS             5288..5728
                     /codon_start=1
                     /gene="traJ"
                     /product="plasmid conjugative transfer protein"
                     /label=traJ
                     /protein_id="AFA34964.1"
                     /translation="MLNGNPALRGWPATAGVTDEGKRMADETKPTRKGSPPIKVYCLPD
                     ERRAIEEKAAAAGMSLSAYLLAVGQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWL
                     TDDPRTARFGDATILALLAKIEEKQDELGKVMMGVVRPRAEP"
     gene            5288..5728
                     /gene="traJ"
                     /label=traJ
     CDS             5357..5728
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"