LOCUS       Exported                4353 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Integration vector pMTL-JH2, complete sequence.
ACCESSION   HQ875749
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4353)
  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 4353)
  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 4353)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4353
                     /organism="Integration vector pMTL-JH2"
                     /mol_type="other DNA"
                     /db_xref="taxon:1128731"
     misc_feature    9..306
                     /gene="pyrF fragment"
                     /note="mediates homologous recombination with the 
                     chromosome of Clostridium acetobutylicum ATCC824"
     gene            9..306
                     /gene="pyrF fragment"
                     /label=pyrF fragment
     misc_feature    330..593
                     /label=multiple cloning site
                     /note="multiple cloning site"
     CDS             330..593
                     /codon_start=1
                     /gene="lacZ"
                     /product="beta-galactosidase alpha subunit"
                     /label=lacZ
                     /protein_id="AFA34924.1"
                     /translation="MTMITNSSSVPGDPLESTSRVHGDLEACRHASLALAVVLQRRDWE
                     NPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWR"
     gene            330..593
                     /gene="lacZ"
                     /label=lacZ
     primer_bind     complement(431..448)
                     /label=M13F
                     /note="M13F"
     primer_bind     complement(431..447)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    595..1794
                     /note="mediates homologous recombination with the 
                     chromosome of Clostridium acetobutylicum ATCC824"
     CDS             612..1352
                     /codon_start=1
                     /gene="pyrZ"
                     /product="dihydroorotate dehydrogenase electron transfer 
                     subunit"
                     /label=pyrZ
                     /protein_id="AFA34927.1"
                     /translation="MKEKYTVEKVYENIKVEDGIYKLSIKGEFEVRPGQFYLLRAWDIE
                     PTLSRPISIYDADDEKISFLYSVVGKGTEILSKLKSGDEIKITGPLGNGFNVKRISGKV
                     AIVCGGIGVAPMVYLAKNLKNCNVDFYAGFKTVSKTVDNVEKYVKELKLSTEDGSIGHK
                     GYVTDNFKPEEYDYVLCCGPEIMMYKVVKMCEQKNVPVYISMEKKMACGIGACLVCTCK
                     TKGGRRRACKEGPVFLGSELILND"
     gene            612..1352
                     /gene="pyrZ"
                     /label=pyrZ
     gene            1345..1794
                     /gene="pyrD fragment"
                     /label=pyrD fragment
     CDS             2173..2613
                     /codon_start=1
                     /gene="repL"
                     /product="plasmid replication protein"
                     /label=repL
                     /protein_id="AFA34925.1"
                     /translation="MKERYGTVYKGSQRLIDEESGEVIEVDKLYRKQTSGNFVKAYIVQ
                     LISMLDMIGGKKLKIVNYILDNVHLSNNTMIATTREIAKATGTSLQTVITTLKILEEGN
                     IIKRKTGVLMLNPELLMRGDDQKQKYLLLEFGNFEQEANEID"
     gene            2173..2613
                     /gene="repL"
                     /label=repL
     CDS             2763..3386
                     /codon_start=1
                     /gene="catP"
                     /product="chloramphenicol acetyltransferase"
                     /label=catP
                     /protein_id="AFA34926.1"
                     /translation="MVFEKIDKNSWNRKEYFDHYFASVPCTYSMTVKVDITQIKEKGMK
                     LYPAMLYYIAMIVNRHSEFRTAINQDGELGIYDEMIPSYTIFHNDTETFSSLWTECKSD
                     FKSFLADYESDTQRYGNNHRMEGKPNAPENIFNVSMIPWSTFDGFNLNLQKGYDYLIPI
                     FTMGKYYKEDNKIILPLAIQVHHAVCDGFHICRFVNELQELINS"
     gene            2763..3386
                     /gene="catP"
                     /label=catP
     rep_origin      3563..4151
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      3599..4144
                     /label=ColE1 RNA II
                     /note="ColE1 RNA II"