LOCUS       Exported                4034 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Integration vector pMTL-JH15, complete sequence.
ACCESSION   HQ875756
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4034)
  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 4034)
  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 4034)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4034
                     /organism="Integration vector pMTL-JH15"
                     /mol_type="other DNA"
                     /db_xref="taxon:1128726"
     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             413..1150
                     /codon_start=1
                     /gene="ermB"
                     /product="MLS resistance protein"
                     /label=ermB
                     /protein_id="AFA34947.1"
                     /translation="MNKNIKYSQNFLTSEKVLNQIIKQLNLKETDTVYEIGTGKGHLTT
                     KLAKISKQVTSIELDSHLFNLSSEKLKLNTRVTLIHQDILQFQFPNKQRYKIVGSIPYH
                     LSTQIIKKVVFESHASDIYLIVEEGFYKRTLDIHRTLGLLLHTQVSIQQLLKLPAECFH
                     PKPKVNSVLIKLTRHTTDVPDKYWKLYTYFVSKWVNREYRQLFTKNQFHQAMKHAKVNN
                     LSTVTYEQVLSIFNSYLLFNGRK"
     gene            413..1150
                     /gene="ermB"
                     /label=ermB
     misc_feature    1169..1432
                     /label=multiple cloning site
                     /note="multiple cloning site"
     CDS             1169..1432
                     /codon_start=1
                     /gene="lacZ"
                     /product="beta-galactosidase alpha subunit"
                     /label=lacZ
                     /protein_id="AFA34948.1"
                     /translation="MTMITNSSSVPGDPLESTSRVHGDLEACRHASLALAVVLQRRDWE
                     NPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWR"
     gene            1169..1432
                     /gene="lacZ"
                     /label=lacZ
     primer_bind     complement(1270..1287)
                     /label=M13F
                     /note="M13F"
     primer_bind     complement(1270..1286)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      1434..1475
                     /regulatory_class="terminator"
                     /note="derived from fdx gene of Clostridium pasteurianum"
     CDS             1854..2294
                     /codon_start=1
                     /gene="repL"
                     /product="plasmid replication protein"
                     /label=repL
                     /protein_id="AFA34949.1"
                     /translation="MKERYGTVYKGSQRLIDEESGEVIEVDKLYRKQTSGNFVKAYIVQ
                     LISMLDMIGGKKLKIVNYILDNVHLSNNTMIATTREIAKATGTSLQTVITTLKILEEGN
                     IIKRKTGVLMLNPELLMRGDDQKQKYLLLEFGNFEQEANEID"
     gene            1854..2294
                     /gene="repL"
                     /label=repL
     CDS             2444..3067
                     /codon_start=1
                     /gene="catP"
                     /product="chloramphenicol acetyltransferase"
                     /label=catP
                     /protein_id="AFA34950.1"
                     /translation="MVFEKIDKNSWNRKEYFDHYFASVPCTYSMTVKVDITQIKEKGMK
                     LYPAMLYYIAMIVNRHSEFRTAINQDGELGIYDEMIPSYTIFHNDTETFSSLWTECKSD
                     FKSFLADYESDTQRYGNNHRMEGKPNAPENIFNVSMIPWSTFDGFNLNLQKGYDYLIPI
                     FTMGKYYKEDNKIILPLAIQVHHAVCDGFHICRFVNELQELINS"
     gene            2444..3067
                     /gene="catP"
                     /label=catP
     rep_origin      3244..3832
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      3280..3825
                     /label=ColE1 RNA II
                     /note="ColE1 RNA II"