LOCUS       V004434                 5115 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V004434
VERSION     V004434
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)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5115)
  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..5115
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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"
     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..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"
     oriT            4468..4577
                     /label="oriT"
                     /note="incP origin of transfer"
     CDS             4610..4978
                     /label="traJ"
                     /note="oriT-recognizing protein"
ORIGIN
        1 cctgcaggag taatataaat gttatagata tattaaaaga atcaaatgca ttattagaag
       61 gtcatttctt actatcatct ggaagacata gtaatagata ttgtcaatgt gcgaaactac
      121 ttcaatatcc acaaaaagca gaaaaagtta ttagtgtaat agcacagaag cttaaaaatg
      181 tagactttaa tataatagta ggaccagcta tgggtggtgt aatagtatcc tatgaattag
      241 caaggcaaac taatagacca ggcatatttg cagaaagaaa agatggaatc atgtgtatac
      301 gaagaggagc 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 atttaaaaat
      601 aaggagtgtc tcaaaataga tttaattttg agacattttt ttagttttaa caaatattaa
      661 acaaaaactt aattttactt taataattat gaaaatggtt attaattttt attgttttga
      721 gttataatag tgttaattat atattataat aattgataca ttttatttat tttagtatag
      781 ataggtatta tatataggag gaattattgt ggcaaaacca agtattttta gcaaagatta
      841 tgaaaataga atgaaaaaga gaaaaagaag gacatttttt tcagttatag ttataatagt
      901 aattttttta ggaataatat tcactaataa tggtattggt aaaaaaataa aaaatagttt
      961 aaatcaaata aaagaagaaa ctaaggctga agaagaacaa aaaaataaac aacaagaaca
     1021 aaaggaaaat aaaaaagaaa atcaaaataa aaatgcagcc acagtaaaaa aagaaagtga
     1081 agttcctaaa gataataata tagaagttca attagaaggc aatataaaaa taaagttgat
     1141 atatatagaa gatagtaata aaaataagac tattaaatct atagatctaa ataaaaataa
     1201 tttattatat gatataaatc cttcaaaaaa ttcaatagtt gtaacaaatc ctaaaactca
     1261 aaatatttat ttagtaaatt taaatggtga aaagcaagat ataacaaata agcagtatac
     1321 atcaacatct ggaacaactt ttcaaaagga tgctatatta gcgtcaaaac aagattattt
     1381 atggggagtt ttacctaagt tcatagataa tgataatata gcctatgttt ctcaactgcc
     1441 atggtttaat aaaaccacta aatatgtatg gatgtataat ataaaaaata aaactcattt
     1501 atataatgaa aatatatcag gagaagacat taagtttgat aaattaacag aaaaaggtct
     1561 tacagtaatt gcagatggca aaactttatt tttaaaagca gatggaagta tagctcagta
     1621 aattaaaaaa tattaaatat aaaagttaat aagtagcaat taaaatataa aatttttaag
     1681 agtgtatgaa atatatacac tcttgcttta tttaaggagg taaatatgga aataggaatg
     1741 tcttctgctt gtttgtatcc taaagtacca atagaagaaa gtatatcagt 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 gcagggcccc ctgcttcggg gtcattatag cgattttttc
     4381 ggtatatcca tcctttttcg cacgatatac aggattttgc caaagggttc gtgtagactt
     4441 tccttggtgt atccaacggc gtcagccggg caggataggt gaagtaggcc cacccgcgag
     4501 cgggtgttcc ttcttcactg tcccttattc gcacctggcg gtgctcaacg ggaatcctgc
     4561 tctgcgaggc tggccggcta ccgccggcgt aacagatgag ggcaagcgga tggctgatga
     4621 aaccaagcca accaggaagg gcagcccacc tatcaaggtg tactgccttc cagacgaacg
     4681 aagagcgatt gaggaaaagg cggcggcggc cggcatgagc ctgtcggcct acctgctggc
     4741 cgtcggccag ggctacaaaa tcacgggcgt cgtggactat gagcacgtcc gcgagctggc
     4801 ccgcatcaat ggcgacctgg gccgcctggg cggcctgctg aaactctggc tcaccgacga
     4861 cccgcgcacg gcgcggttcg gtgatgccac gatcctcgcc ctgctggcga agatcgaaga
     4921 gaagcaggac gagcttggca aggtcatgat gggcgtggtc cgcccgaggg cagagccatg
     4981 acttttttag ccgctaaaac ggccgggggg tgcgcgtgat tgccaagcac gtccccatgc
     5041 gctccatcaa gaagagcgac ttcgcggagc tggtgaagta catcaccgac gagcaaggca
     5101 agaccgatcg ggccc
//
