LOCUS       Exported                5115 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Integration vector pMTL-JH27, complete sequence.
ACCESSION   HQ875763
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5115)
  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 5115)
  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 5115)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5115
                     /organism="Integration vector pMTL-JH27"
                     /mol_type="other DNA"
                     /db_xref="taxon:1128734"
     misc_feature    9..308
                     /gene="pyrE fragment"
                     /note="mediates homologous recombination with the 
                     chromosome of Clostridium sporogenes NCIMB10696"
     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="AFA34978.1"
                     /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 sporogenes NCIMB10696"
     CDS             809..1621
                     /codon_start=1
                     /product="hypothetical protein"
                     /label=hypothetical protein
                     /note="CBO3234-like"
                     /protein_id="AFA34979.1"
                     /translation="MAKPSIFSKDYENRMKKRKRRTFFSVIVIIVIFLGIIFTNNGIGK
                     KIKNSLNQIKEETKAEEEQKNKQQEQKENKKENQNKNAATVKKESEVPKDNNIEVQLEG
                     NIKIKLIYIEDSNKNKTIKSIDLNKNNLLYDINPSKNSIVVTNPKTQNIYLVNLNGEKQ
                     DITNKQYTSTSGTTFQKDAILASKQDYLWGVLPKFIDNDNIAYVSQLPWFNKTTKYVWM
                     YNIKNKTHLYNENISGEDIKFDKLTEKGLTVIADGKTLFLKADGSIAQ"
     RBS             1713..1721
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3 end of the 16S rRNA (Chen et al., 1994)"
     CDS             2169..2609
                     /codon_start=1
                     /gene="repL"
                     /product="plasmid replication protein"
                     /label=repL
                     /protein_id="AFA34980.1"
                     /translation="MKERYGTVYKGSQRLIDEESGEVIEVDKLYRKQTSGNFVKAYIVQ
                     LISMLDMIGGKKLKIVNYILDNVHLSNNTMIATTREIAKATGTSLQTVITTLKILEEGN
                     IIKRKTGVLMLNPELLMRGDDQKQKYLLLEFGNFEQEANEID"
     gene            2169..2609
                     /gene="repL"
                     /label=repL
     CDS             2759..3382
                     /codon_start=1
                     /gene="catP"
                     /product="chloramphenicol acetyltransferase"
                     /label=catP
                     /protein_id="AFA34981.1"
                     /translation="MVFEKIDKNSWNRKEYFDHYFASVPCTYSMTVKVDITQIKEKGMK
                     LYPAMLYYIAMIVNRHSEFRTAINQDGELGIYDEMIPSYTIFHNDTETFSSLWTECKSD
                     FKSFLADYESDTQRYGNNHRMEGKPNAPENIFNVSMIPWSTFDGFNLNLQKGYDYLIPI
                     FTMGKYYKEDNKIILPLAIQVHHAVCDGFHICRFVNELQELINS"
     gene            2759..3382
                     /gene="catP"
                     /label=catP
     rep_origin      3559..4147
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      3595..4140
                     /label=ColE1 RNA II
                     /note="ColE1 RNA II"
     oriT            4468..4577
                     /note="incP origin of transfer"
     CDS             4541..4981
                     /codon_start=1
                     /gene="traJ"
                     /product="plasmid conjugative transfer protein"
                     /label=traJ
                     /protein_id="AFA34982.1"
                     /translation="MLNGNPALRGWPATAGVTDEGKRMADETKPTRKGSPPIKVYCLPD
                     ERRAIEEKAAAAGMSLSAYLLAVGQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWL
                     TDDPRTARFGDATILALLAKIEEKQDELGKVMMGVVRPRAEP"
     gene            4541..4981
                     /gene="traJ"
                     /label=traJ
     CDS             4610..4981
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"