LOCUS       Exported                3870 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Integration vector pMTL-JH30, complete sequence.
ACCESSION   HQ875766
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3870)
  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 3870)
  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 3870)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..3870
                     /organism="Integration vector pMTL-JH30"
                     /mol_type="other DNA"
                     /db_xref="taxon:1128738"
     regulatory      9..62
                     /regulatory_class="terminator"
                     /note="derived from CD0164 gene of Clostridium difficile"
     primer_bind     63..80
                     /label=M13R
                     /note="M13R"
     primer_bind     63..79
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    90..389
                     /gene="thl fragment"
                     /note="mediates homologous recombination with the 
                     chromosome of Clostridium acetobutylicum ATCC824"
     gene            90..389
                     /gene="thl fragment"
                     /label=thl fragment
     CDS             412..987
                     /codon_start=1
                     /gene="pyrE"
                     /product="orotate phosphoribosyltransferase"
                     /label=pyrE
                     /protein_id="AFA34992.1"
                     /translation="MSNINVIDILKESNALLEGHFLLSSGRHSNRYCQCAKLLQYPQKA
                     EKVISVVAEKLKDADFNIIVGPAMGGVIVSYELARQTNRPGIFAERKDGIMCIRRGFEI
                     KKGDKVIISEDVITTGKSSLEVAKVIEEMGGEVVGIACIVDRRAEGIKTNYPIYSACKL
                     EIETYEKDNCELCKKNIPFVKPGSREQK"
     gene            412..987
                     /gene="pyrE"
                     /label=pyrE
     misc_feature    1005..1268
                     /label=multiple cloning site
                     /note="multiple cloning site"
     CDS             1005..1268
                     /codon_start=1
                     /gene="lacZ"
                     /product="beta-galactosidase alpha subunit"
                     /label=lacZ
                     /protein_id="AFA34993.1"
                     /translation="MTMITNSSSVPGDPLESTSRVHGDLEACRHASLALAVVLQRRDWE
                     NPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWR"
     gene            1005..1268
                     /gene="lacZ"
                     /label=lacZ
     primer_bind     complement(1106..1123)
                     /label=M13F
                     /note="M13F"
     primer_bind     complement(1106..1122)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      1270..1311
                     /regulatory_class="terminator"
                     /note="derived from fdx gene of Clostridium pasteurianum"
     CDS             1690..2130
                     /codon_start=1
                     /gene="repL"
                     /product="plasmid replication protein"
                     /label=repL
                     /protein_id="AFA34994.1"
                     /translation="MKERYGTVYKGSQRLIDEESGEVIEVDKLYRKQTSGNFVKAYIVQ
                     LISMLDMIGGKKLKIVNYILDNVHLSNNTMIATTREIAKATGTSLQTVITTLKILEEGN
                     IIKRKTGVLMLNPELLMRGDDQKQKYLLLEFGNFEQEANEID"
     gene            1690..2130
                     /gene="repL"
                     /label=repL
     CDS             2280..2903
                     /codon_start=1
                     /gene="catP"
                     /product="chloramphenicol acetyltransferase"
                     /label=catP
                     /protein_id="AFA34995.1"
                     /translation="MVFEKIDKNSWNRKEYFDHYFASVPCTYSMTVKVDITQIKEKGMK
                     LYPAMLYYIAMIVNRHSEFRTAINQDGELGIYDEMIPSYTIFHNDTETFSSLWTECKSD
                     FKSFLADYESDTQRYGNNHRMEGKPNAPENIFNVSMIPWSTFDGFNLNLQKGYDYLIPI
                     FTMGKYYKEDNKIILPLAIQVHHAVCDGFHICRFVNELQELINS"
     gene            2280..2903
                     /gene="catP"
                     /label=catP
     rep_origin      3080..3668
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      3116..3661
                     /label=ColE1 RNA II
                     /note="ColE1 RNA II"