LOCUS       V004445                 4349 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V004445
VERSION     V004445
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 4349)
  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 4349)
  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 4349)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4349)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic
            Acids Res."; date: "2012"; volume: "40"; issue: "8"; pages: "E59"
            SGRef: number: 2; type: "Journal Article"; journalName: "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"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4349
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    9..308
                     /gene="pyrE fragment"
                     /note="mediates homologous recombination with the
                     chromosome of Clostridium acetobutylicum ATCC824"
     gene            9..308
                     /gene="pyrE fragment"
                     /label="pyrE fragment"
     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 acetobutylicum ATCC824"
     gene            complement(641..1790)
                     /gene="hydA fragment"
                     /label="hydA fragment"
     CDS             2169..2609
                     /codon_start=1
                     /gene="repL"
                     /product="plasmid replication protein"
                     /label="repL"
                     /protein_id="AFA34939.1"
                     /translation="MKERYGTVYKGSQRLIDEESGEVIEVDKLYRKQTSGNFVKAYIVQ
                     LISMLDMIGGKKLKIVNYILDNVHLSNNTMIATTREIAKATGTSLQTVITTLKILEEGN
                     IIKRKTGVLMLNPELLMRGDDQKQKYLLLEFGNFEQEANEID"
     gene            2169..2609
                     /gene="repL"
                     /label="repL"
     CDS             2759..3379
                     /gene="catP"
                     /label="Chloramphenicol acetyltransferase"
                     /note="Chloramphenicol acetyltransferase from Clostridium
                     perfringens. Accession#: P26826"
     rep_origin      3559..4147
                     /direction=RIGHT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
ORIGIN
        1 cctgcaggag agtaatgtac ttacctttgg ggatttcata actaaaagcg gcagaagaac
       61 accatttttt ataaatacag gtaactacaa gacaggtaat caattaaata agttggctaa
      121 gttttatgct aaagcaatat atgataattt tggagatgat tttgatattt tatttgggcc
      181 tgcatataaa ggaatacctt taagtgtttc agtagctatg gcacttgata atatttatgg
      241 aattaatgca gcttattgtt caaatagaaa agaagttaaa gatcacggtg ataagggaat
      301 acttcttggc ggccgctgta tccatatgac catgattacg aattcgagct cggtacccgg
      361 ggatcctcta gagtcgacgt cacgcgtcca tggagatctc gaggcctgca gacatgcaag
      421 cttggcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact
      481 taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac
      541 cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggcgctagc taaaataaat
      601 gtgcctcaac ttagatgtta aggcacattt attttatata ttattcatgt tttgaaacat
      661 ttttatcttt tgtgtatttt acgtgtagta atttgtgagc aagtccttca cctggttttc
      721 caaagtagct atcatacatt ttaataatag ctggattatc atgtgacttt ctctttgaaa
      781 gaacattttt atcttggttg tataatactg atgctcttag ttttctgtaa tcaacatttt
      841 ctctatcaag agcatttacg tgaggttgac ctccaccatt tatacatcca ccagggcaag
      901 ccattacttc tataaagtga tattgttttt cgttcatttt tccagatttc ataaactcga
      961 agaagttaga agcaccattt ataacagcaa cgtttagttt atttccagca atttcaactt
     1021 ccgcttcttt tatgccttta aagcctctta cttcagtgta atcaacattt tcaagttctt
     1081 tattttcagc aaagtcttta gctgatctta ttgcagcttc cataacgcca ccggttgcac
     1141 caaagatagc tccagcacca ctgtaagtac ccatagcagg atcaacttca ccatcttcaa
     1201 gatctgcaaa tttaattttt gcatctttaa tcatttttgc aagctctctt gtagttaagg
     1261 atgcatcaat atctcttaag ctgttagttt ccatgaaagg aatatctgct tcatattttt
     1321 tatcattaca aggcatgata gtaactgtat aaacatcttc tggagctatt cctgaaattg
     1381 aaggatagta agtttttgat gcagtaccaa atatttgttg tggtgatttt gctgatgaaa
     1441 gattatctaa taattcagga tgataatttt gagctaatct tacccatgca ggacagcaag
     1501 atgtaaacat agggaatggg ccattatttt taactctgcc taaaagttca gtagcttctt
     1561 ccattatagt catatctgca ccaaagttta tatcaaatac tttatcaaag cctaacattc
     1621 taagtgcagt atatagtttt cctgttacat cttttccata tcccattttg aataattcgc
     1681 ccatagcagt tcttactgat ggagccattg caacaatgac atgtttttta gggtcattaa
     1741 gagcttcttg aactttttct atatgggatt tttcttttaa agcagcaaca ggcgcgccgc
     1801 attcacttct tttctatata aatatgagcg aagcgaataa gcgtcggaaa agcagcaaaa
     1861 agtttccttt ttgctgttgg agcatggggg ttcagggggt gcagtatctg acgtcaatgc
     1921 cgagcgaaag cgagccgaag ggtagcattt acgttagata accccctgat atgctccgac
     1981 gctttatata gaaaagaaga ttcaactagg taaaatctta atataggttg agatgataag
     2041 gtttataagg aatttgtttg ttctaatttt tcactcattt tgttctaatt tcttttaaca
     2101 aatgttcttt tttttttaga acagttatga tatagttaga atagtttaaa ataaggagtg
     2161 agaaaaagat gaaagaaaga tatggaacag tctataaagg ctctcagagg ctcatagacg
     2221 aagaaagtgg agaagtcata gaggtagaca agttataccg taaacaaacg tctggtaact
     2281 tcgtaaaggc atatatagtg caattaataa gtatgttaga tatgattggc ggaaaaaaac
     2341 ttaaaatcgt taactatatc ctagataatg tccacttaag taacaataca atgatagcta
     2401 caacaagaga aatagcaaaa gctacaggaa caagtctaca aacagtaata acaacactta
     2461 aaatcttaga agaaggaaat attataaaaa gaaaaactgg agtattaatg ttaaaccctg
     2521 aactactaat gagaggcgac gaccaaaaac aaaaatacct cttactcgaa tttgggaact
     2581 ttgagcaaga ggcaaatgaa atagattgac ctcccaataa caccacgtag ttattgggag
     2641 gtcaatctat gaaatgcgat taagggccgg ccagtgggca agttgaaaaa ttcacaaaaa
     2701 tgtggtataa tatctttgtt cattagagcg ataaacttga atttgagagg gaacttagat
     2761 ggtatttgaa aaaattgata aaaatagttg gaacagaaaa gagtattttg accactactt
     2821 tgcaagtgta ccttgtacct acagcatgac cgttaaagtg gatatcacac aaataaagga
     2881 aaagggaatg aaactatatc ctgcaatgct ttattatatt gcaatgattg taaaccgcca
     2941 ttcagagttt aggacggcaa tcaatcaaga tggtgaattg gggatatatg atgagatgat
     3001 accaagctat acaatatttc acaatgatac tgaaacattt tccagccttt ggactgagtg
     3061 taagtctgac tttaaatcat ttttagcaga ttatgaaagt gatacgcaac ggtatggaaa
     3121 caatcataga atggaaggaa agccaaatgc tccggaaaac atttttaatg tatctatgat
     3181 accgtggtca accttcgatg gctttaatct gaatttgcag aaaggatatg attatttgat
     3241 tcctattttt actatgggga aatattataa agaagataac aaaattatac ttcctttggc
     3301 aattcaagtt catcacgcag tatgtgacgg atttcacatt tgccgttttg taaacgaatt
     3361 gcaggaattg ataaatagtt aacttcaggt ttgtctgtaa ctaaaaacaa gtatttaagc
     3421 aaaaacatcg tagaaatacg gtgttttttg ttaccctaag tttaaactcc tttttgataa
     3481 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     3541 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     3601 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     3661 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     3721 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     3781 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     3841 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     3901 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     3961 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     4021 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     4081 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     4141 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     4201 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     4261 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     4321 agcggaagag cgcccaatac gcagggccc
//
