LOCUS       V015376                 7558 bp    DNA     circular SYN 16-MAY-2020
DEFINITION  Exported.
ACCESSION   V015376
VERSION     V015376
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 7558)
  AUTHORS   Guo L.
  TITLE     Direct Submission
  JOURNAL   Submitted (28-OCT-2019) Jiangnan University, State Key Laboratory of
            Food Science and Technology, 1800 Lihu Avenue, Wuxi, Jiangsu 214122,
            China
REFERENCE   2  (bases 1 to 7558)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Submitted
            (28-OCT-2019) Jiangnan University, State Key Laboratory of Food
            Science and Technology, 1800 Lihu Avenue, Wuxi, Jiangsu 214122,
            China"
FEATURES             Location/Qualifiers
     source          1..7558
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      12..467
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             complement(563..1375)
                     /label="KanR"
                     /note="aminoglycoside phosphotransferase"
     rep_origin      1497..2085
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     misc_feature    complement(2271..2413)
                     /label="bom"
                     /note="basis of mobility region from pBR322"
     CDS             complement(2518..2706)
                     /label="rop"
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     protein_bind    complement(3481..3502)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(3518..4597)
                     /label="lacI"
                     /note="lac repressor"
     promoter        complement(4598..4675)
                     /label="lacI promoter"
     promoter        4984..5002
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5003..5027
                     /label="lac operator"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             5042..5064
                     /label="RBS"
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5083..5100
                     /label="6xHis"
                     /note="6xHis affinity tag"
     CDS             5110..5127
                     /label="thrombin site"
                     /note="thrombin recognition and cleavage site"
     CDS             5131..5163
                     /label="T7 tag (gene 10 leader)"
                     /note="leader peptide from bacteriophage T7 gene 10"
     promoter        5357..5385
                     /label="tac promoter"
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     CDS             5432..6610
                     /gene="phaA"
                     /label="Acetyl-CoA acetyltransferase"
                     /note="Acetyl-CoA acetyltransferase from Cupriavidus
                     necator (strain ATCC 17699 / DSM 428 / KCTC 22496 / NCIMB
                     10442 / H16 / Stanier 337). Accession#: P14611"
     CDS             6639..7376
                     /gene="phaB"
                     /label="Acetoacetyl-CoA reductase"
                     /note="Acetoacetyl-CoA reductase from Cupriavidus necator
                     (strain ATCC 17699 / DSM 428 / KCTC 22496 / NCIMB 10442 /
                     H16 / Stanier 337). Accession#: P14697"
     CDS             7402..7419
                     /label="6xHis"
                     /note="6xHis affinity tag"
     terminator      7486..7533
                     /label="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA
                     polymerase"
ORIGIN
        1 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
       61 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
      121 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
      181 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
      241 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
      301 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
      361 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
      421 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaatttcag gtggcacttt
      481 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
      541 tccgctcatg aattaattct tagaaaaact catcgagcat caaatgaaac tgcaatttat
      601 tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa
      661 actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc
      721 gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga
      781 aatcaccatg agtgacgact gaatccggtg agaatggcaa aagtttatgc atttctttcc
      841 agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac
      901 cgttattcat tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac
      961 aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat
     1021 tttcacctga atcaggatat tcttctaata cctggaatgc tgttttcccg gggatcgcag
     1081 tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca
     1141 taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac
     1201 ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaat cgatagattg
     1261 tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca
     1321 tgttggaatt taatcgcggc ctagagcaag acgtttcccg ttgaatatgg ctcataacac
     1381 cccttgtatt actgtttatg taagcagaca gttttattgt tcatgaccaa aatcccttaa
     1441 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     1501 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     1561 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     1621 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
     1681 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     1741 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     1801 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     1861 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     1921 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     1981 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     2041 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     2101 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     2161 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     2221 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg
     2281 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatatggtgc actctcagta
     2341 caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg
     2401 ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     2461 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     2521 gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc
     2581 gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag
     2641 aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt
     2701 ggtcactgat gcctccgtgt aagggggatt tctgttcatg ggggtaatga taccgatgaa
     2761 acgagagagg atgctcacga tacgggttac tgatgatgaa catgcccggt tactggaacg
     2821 ttgtgagggt aaacaactgg cggtatggat gcggcgggac cagagaaaaa tcactcaggg
     2881 tcaatgccag cgcttcgtta atacagatgt aggtgttcca cagggtagcc agcagcatcc
     2941 tgcgatgcag atccggaaca taatggtgca gggcgctgac ttccgcgttt ccagacttta
     3001 cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg ttttgcagca
     3061 gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc tgctaaccag taaggcaacc
     3121 ccgccagcct agccgggtcc tcaacgacag gagcacgatc atgcgcaccc gtggggccgc
     3181 catgccggcg ataatggcct gcttctcgcc gaaacgtttg gtggcgggac cagtgacgaa
     3241 ggcttgagcg agggcgtgca agattccgaa taccgcaagc gacaggccga tcatcgtcgc
     3301 gctccagcga aagcggtcct cgccgaaaat gacccagagc gctgccggca cctgtcctac
     3361 gagttgcatg ataaagaaga cagtcataag tgcggcgacg atagtcatgc cccgcgccca
     3421 ccggaaggag ctgactgggt tgaaggctct caagggcatc ggtcgagatc ccggtgccta
     3481 atgagtgagc taacttacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     3541 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     3601 tgggcgccag ggtggttttt cttttcacca gtgagacggg caacagctga ttgcccttca
     3661 ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa
     3721 aatcctgttt gatggtggtt aacggcggga tataacatga gctgtcttcg gtatcgtcgt
     3781 atcccactac cgagatatcc gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg
     3841 cgcccagcgc catctgatcg ttggcaacca gcatcgcagt gggaacgatg ccctcattca
     3901 gcatttgcat ggtttgttga aaaccggaca tggcactcca gtcgccttcc cgttccgcta
     3961 tcggctgaat ttgattgcga gtgagatatt tatgccagcc agccagacgc agacgcgccg
     4021 agacagaact taatgggccc gctaacagcg cgatttgctg gtgacccaat gcgaccagat
     4081 gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat aatactgttg atgggtgtct
     4141 ggtcagagac atcaagaaat aacgccggaa cattagtgca ggcagcttcc acagcaatgg
     4201 catcctggtc atccagcgga tagttaatga tcagcccact gacgcgttgc gcgagaagat
     4261 tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc taccatcgac accaccacgc
     4321 tggcacccag ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca
     4381 gggccagact ggaggtggca acgccaatca gcaacgactg tttgcccgcc agttgttgtg
     4441 ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc ttccactttt tcccgcgttt
     4501 tcgcagaaac gtggctggcc tggttcacca cgcgggaaac ggtctgataa gagacaccgg
     4561 catactctgc gacatcgtat aacgttactg gtttcacatt caccaccctg aattgactct
     4621 cttccgggcg ctatcatgcc ataccgcgaa aggttttgcg ccattcgatg gtgtccggga
     4681 tctcgacgct ctcccttatg cgactcctgc attaggaagc agcccagtag taggttgagg
     4741 ccgttgagca ccgccgccgc aaggaatggt gcatgcaagg agatggcgcc caacagtccc
     4801 ccggccacgg ggcctgccac catacccacg ccgaaacaag cgctcatgag cccgaagtgg
     4861 cgagcccgat cttccccatc ggtgatgtcg gcgatatagg cgccagcaac cgcacctgtg
     4921 gcgccggtga tgccggccac gatgcgtccg gcgtagagga tcgagatctc gatcccgcga
     4981 aattaatacg actcactata ggggaattgt gagcggataa caattcccct ctagaaataa
     5041 ttttgtttaa ctttaagaag gagatatacc atgggcagca gccatcatca tcatcatcac
     5101 agcagcggcc tggtgccgcg cggcagccat atggctagca tgactggtgg acagcaaatg
     5161 ggtcgcggat ccggagctta tcgactgcac ggtgcaccaa tgcttctggc gtcaggcagc
     5221 catcggaagc tgtggtatgg ctgtgcaggt cgtaaatcac tgcataattc gtgtcgctca
     5281 aggcgcactc ccgttctgga taatgttttt tgcgccgaca tcataacggt tctggcaaat
     5341 attctgaaat gagctgttga caattaatca tcggctcgta taatgtgtgg aattgtgagc
     5401 ggataacaat ttcacacagg aaacagaatt catgacagac gtggtgattg ttagcgcagc
     5461 ccgcaccgcc gttggtaagt ttggcggcag cctggcaaag atccctgccc ctgagctggg
     5521 cgcagtggtt atcaaggcag ccctggagcg cgcaggtgtg aaaccggagc aggtgagtga
     5581 ggtgatcatg ggccaggttc tgaccgcagg tagtggccaa aatccggccc gtcaggccgc
     5641 aatcaaagca ggcttacctg ccatggtgcc ggccatgacc atcaacaagg tgtgcggcag
     5701 tggcctgaag gccgtgatgt tagccgccaa cgccatcatg gccggcgatg cagagatcgt
     5761 ggtggcaggc ggccaggaaa atatgagcgc cgcacctcat gttctgccgg gcagtcgcga
     5821 tggcttccgt atgggcgacg caaagctggt ggacaccatg atcgtggacg gcctgtggga
     5881 cgtgtacaac cagtaccaca tgggcatcac cgccgagaac gtggccaagg agtacggcat
     5941 cacacgcgag gcccaggatg agttcgcagt tggcagccag aacaaggcag aggccgccca
     6001 gaaagccggc aagttcgatg aagaaatcgt gccggtgctg atcccgcagc gcaaaggtga
     6061 cccggtggcc ttcaaaaccg atgagttcgt gcgccagggc gcaaccctgg atagcatgag
     6121 cggcctgaag ccggccttcg ataaggcagg cacagtgacc gccgccaatg ccagtggcct
     6181 gaacgatggt gcagccgcag tggttgtgat gagcgccgcc aaagccaagg aactgggtct
     6241 gaccccgctg gcaaccatca aaagctacgc caacgccggc gttgatccta aagtgatggg
     6301 catgggtccg gttccggcca gtaagcgcgc cttaagtcgc gccgaatgga caccgcagga
     6361 cctggacctg atggaaatca acgaagcctt cgcagcccag gcattagcag tgcatcagca
     6421 aatgggctgg gacaccagca aggtgaacgt gaacggtggc gccatcgcaa ttggccaccc
     6481 tatcggcgca agtggctgtc gcatcctggt gaccctgctg cacgaaatga agcgccgtga
     6541 cgccaagaaa ggcctggcca gcttatgcat tggtggtggt atgggcgttg ccttagcagt
     6601 ggaacgcaag taagtcgact cacacaggaa aggagctcat gacccagcgc attgcctatg
     6661 tgaccggtgg tatgggcggt attggtaccg ccatctgtca gcgtctggca aaagatggct
     6721 ttcgcgttgt tgctggttgc ggtccgaata gtccgcgccg tgaaaaatgg ctggaacagc
     6781 agaaggcact gggcttcgac tttatcgcaa gcgaaggcaa tgtggcagac tgggatagca
     6841 ccaaaaccgc cttcgataag gtgaaaagcg aggtgggcga agttgacgtg ctgatcaaca
     6901 acgctggtat cacccgcgat gtggtgtttc gcaaaatgac ccgcgccgat tgggatgcag
     6961 tgatcgatac caatttaacc tctttattca acgtgaccaa acaagttatt gacggcatgg
     7021 cagatcgtgg ctggggtcgc attgttaaca tcagcagcgt gaacggccag aaaggtcagt
     7081 ttggccagac caactacagc accgccaaag ccggtttaca cggttttacc atggctttag
     7141 cacaagaagt ggccaccaaa ggtgtgaccg tgaataccgt tagcccgggc tacattgcca
     7201 ccgacatggt gaaagccatt cgccaagatg ttctggacaa aatcgtggca accatcccgg
     7261 tgaaacgtct gggtttaccg gaagagatcg ccagcatttg cgcttggctg agcagtgaag
     7321 aaagcggctt cagcaccggt gccgatttct ctttaaacgg tggtttacat atgggttaag
     7381 ttccatccca atggcctcga gcaccaccac caccaccact gagatccggc tgctaacaaa
     7441 gcccgaaagg aagctgagtt ggctgctgcc accgctgagc aataactagc ataacccctt
     7501 ggggcctcta aacgggtctt gaggggtttt ttgctgaaag gaggaactat atccggat
//
