LOCUS       V015977                 5734 bp    DNA     circular SYN 12-DEC-2019
DEFINITION  Exported.
ACCESSION   V015977
VERSION     V015977
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 5734)
  AUTHORS   Hartley CJ, Williams CC, Scoble JA, Churches QI, North A, French NG,
            Nebl T, Coia G, Warden AC, Simpson G, Frazer AR, Nixon Jensen C,
            Turner NJ, Scott C.
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 5734)
  AUTHORS   Hartley CJ, Williams CC, Scoble JA, Churches QI, North A, French NG,
            Nebl T, Coia G, Warden AC, Simpson G, Frazer AR, Nixon Jensen C,
            Turner NJ, Scott C.
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 5734)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Submitted
            (10-MAY-2019) Biocatalysis "
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (12-MAY-2019) Biocatalysis "
FEATURES             Location/Qualifiers
     source          1..5734
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        21..39
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             71..93
                     /label="RBS"
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             113..130
                     /label="6xHis"
                     /note="6xHis affinity tag"
     protein_bind    140..164
                     /label="attB1"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             170..1186
                     /gene="gpsA"
                     /label="Glycerol-3-phosphate dehydrogenase [NAD(P)+]"
                     /note="Glycerol-3-phosphate dehydrogenase [NAD(P)+] from
                     Escherichia coli (strain SMS-3-5 / SECEC). Accession#:
                     B1LK47"
     protein_bind    complement(1200..1224)
                     /label="attB2"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      1291..1338
                     /label="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA
                     polymerase"
     promoter        complement(1704..1732)
                     /label="tet promoter"
                     /note="E. coli promoter for tetracycline efflux protein
                     gene"
     promoter        1845..1949
                     /label="AmpR promoter"
     CDS             1950..2807
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      2981..3569
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     misc_feature    complement(3755..3897)
                     /label="bom"
                     /note="basis of mobility region from pBR322"
     CDS             complement(4002..4190)
                     /label="rop"
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
ORIGIN
        1 agatctcgat cccgcgaaat taatacgact cactataggg agaccacaac ggtttccctc
       61 tagaaataat tttgtttaac tttaagaagg agatatacat atgtcgtact accatcacca
      121 tcaccatcac ctcgaatcaa caagtttgta caaaaaagca ggcttccata tgaatcagcg
      181 taatgcaagc atgaccgtta ttggtgcagg tagctatggc accgcactgg caattaccct
      241 ggcacgtaat ggtcatgaag ttgttctgtg gggtcatgat ccggaacaca ttgcaaccct
      301 ggaacgtgat cgttgtaatg cagcatttct gccggatgtt ccgtttccgg ataccctgca
      361 tctggaaagc gatctggcaa cagcactggc agcaagccgt aatattctgg ttgttgttcc
      421 gagccatgtt tttggtgaag ttctgcgtca gattaaaccg ctgatgcgtc cggatgcacg
      481 tctggtttgg gcaaccaaag gtctggaagc agaaaccggt cgtctgctgc aggatgttgc
      541 acgtgaagca ctgggtgatc agattccgct ggcagttatt agcggtccga cctttgcaaa
      601 agaactggca gccggtctgc cgaccgccat tagcctggca agcaccgatc agacatttgc
      661 cgatgatctg cagcaactgc tgcattgtgg taaaagcttt cgtgtttata gcaacccgga
      721 ttttattggt gttcagctgg gtggtgcagt gaaaaatgtt attgcaattg gtgccggtat
      781 gagtgatggt attggttttg gtgcaaatgc acgtaccgca ctgattaccc gtggtctggc
      841 cgaaatgagc cgtctgggtg cagccctggg tgccgatccg gcaaccttta tgggtatggc
      901 aggtctgggc gatctggttc tgacctgtac cgataatcag agccgtaatc gtcgttttgg
      961 tatgatgctg ggtcagggta tggatgttca gagcgcacaa gaaaaaattg gtcaggttgt
     1021 tgaaggttac cgcaatacca aagaagtgcg cgaactggca catcgttttg gcgttgaaat
     1081 gccgattacc gaagaaatct atcaggttct gtattgcggc aaaaatgccc gtgaagcagc
     1141 actgaccctg ctgggtcgtg cacgtaaaga tgaacgtagc agccattaat aagaattcga
     1201 cccagctttc ttgtacaaag tggttgattc gaggctgcta acaaagcccg aaaggaagct
     1261 gagttggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg
     1321 gtcttgaggg gttttttgct gaaaggagga actatatccg gatatccaca ggacgggtgt
     1381 ggtcgccatg atcgcgtagt cgatagtggc tccaagtagc gaagcgagca ggactgggcg
     1441 gcggccaaag cggtcggaca gtgctccgag aacgggtgcg catagaaatt gcatcaacgc
     1501 atatagcgct agcagcacgc catagtgact ggcgatgctg tcggaatgga cgatatcccg
     1561 caagaggccc ggcagtaccg gcataaccaa gcctatgcct acagcatcca gggtgacggt
     1621 gccgaggatg acgatgagcg cattgttaga tttcatacac ggtgcctgac tgcgttagca
     1681 atttaactgt gataaactac cgcattaaag cttatcgatg ataagctgtc aaacatgaga
     1741 attcttgaag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa
     1801 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     1861 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     1921 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     1981 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     2041 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     2101 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     2161 aagttctgct atgtggcgcg gtattatccc gtgttgacgc cgggcaagag caactcggtc
     2221 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     2281 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     2341 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     2401 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     2461 taccaaacga cgagcgtgac accacgatgc ctgcagcaat ggcaacaacg ttgcgcaaac
     2521 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     2581 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     2641 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     2701 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     2761 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     2821 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     2881 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     2941 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     3001 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     3061 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     3121 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     3181 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     3241 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     3301 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     3361 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     3421 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     3481 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     3541 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     3601 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     3661 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     3721 gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc
     3781 atctgtgcgg tatttcacac cgcatatatg gtgcactctc agtacaatct gctctgatgc
     3841 cgcatagtta agccagtata cactccgcta tcgctacgtg actgggtcat ggctgcgccc
     3901 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
     3961 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
     4021 ccgaaacgcg cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgaag cgattcacag
     4081 atgtctgcct gttcatccgc gtccagctcg ttgagtttct ccagaagcgt taatgtctgg
     4141 cttctgataa agcgggccat gttaagggcg gttttttcct gtttggtcac tgatgcctcc
     4201 gtgtaagggg gatttctgtt catgggggta atgataccga tgaaacgaga gaggatgctc
     4261 acgatacggg ttactgatga tgaacatgcc cggttactgg aacgttgtga gggtaaacaa
     4321 ctggcggtat ggatgcggcg ggaccagaga aaaatcactc agggtcaatg ccagcgcttc
     4381 gttaatacag atgtaggtgt tccacagggt agccagcagc atcctgcgat gcagatccgg
     4441 aacataatgg tgcagggcgc tgacttccgc gtttccagac tttacgaaac acggaaaccg
     4501 aagaccattc atgttgttgc tcaggtcgca gacgttttgc agcagcagtc gcttcacgtt
     4561 cgctcgcgta tcggtgattc attctgctaa ccagtaaggc aaccccgcca gcctagccgg
     4621 gtcctcaacg acaggagcac gatcatgcgc acccgtggcc aggacccaac gctgcccgag
     4681 atgcgccgcg tgcggctgct ggagatggcg gacgcgatgg atatgttctg ccaagggttg
     4741 gtttgcgcat tcacagttct ccgcaagaat tgattggctc caattcttgg agtggtgaat
     4801 ccgttagcga ggtgccgccg gcttccattc aggtcgaggt ggcccggctc catgcaccgc
     4861 gacgcaacgc ggggaggcag acaaggtata gggcggcgcc tacaatccat gccaacccgt
     4921 tccatgtgct cgccgaggcg gcataaatcg ccgtgacgat cagcggtcca gtgatcgaag
     4981 ttaggctggt aagagccgcg agcgatcctt gaagctgtcc ctgatggtcg tcatctacct
     5041 gcctggacag catggcctgc aacgcgggca tcccgatgcc gccggaagcg agaagaatca
     5101 taatggggaa ggccatccag cctcgcgtcg cgaacgccag caagacgtag cccagcgcgt
     5161 cggccgccat gccggcgata atggcctgct tctcgccgaa acgtttggtg gcgggaccag
     5221 tgacgaaggc ttgagcgagg gcgtgcaaga ttccgaatac cgcaagcgac aggccgatca
     5281 tcgtcgcgct ccagcgaaag cggtcctcgc cgaaaatgac ccagagcgct gccggcacct
     5341 gtcctacgag ttgcatgata aagaagacag tcataagtgc ggcgacgata gtcatgcccc
     5401 gcgcccaccg gaaggagctg actgggttga aggctctcaa gggcatcggt cgatcgacgc
     5461 tctcccttat gcgactcctg cattaggaag cagcccagta gtaggttgag gccgttgagc
     5521 accgccgccg caaggaatgg tgcatgcaag gagatggcgc ccaacagtcc cccggccacg
     5581 gggcctgcca ccatacccac gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga
     5641 tcttccccat cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg
     5701 atgccggcca cgatgcgtcc ggcgtagagg atcg
//
