LOCUS       62056_19380        7358 bp DNA     circular SYN 16-MAY-2020
DEFINITION  Cloning vector pSC101-PhaC-pct, complete sequence.
ACCESSION   MN623110
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7358)
  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 7358)
  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..7358
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(100..129)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     CDS             complement(185..808)
                     /label=TetR
                     /note="tetracycline repressor TetR"
     promoter        824..879
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     gene            909..2588
                     /gene="phaC"
                     /label=phaC
     CDS             909..2588
                     /codon_start=1
                     /transl_table=11
                     /gene="phaC"
                     /product="polyhydroxyalkanoate synthase"
                     /label=phaC
                     /protein_id="QJR97783.1"
                     /translation="MSNKNSDDLNRQASENTLGLNPVIGLRGKDLLTSARMVLTQAIKQ
                     PIHSVKHVAHFGIELKNVMFGKSKLQPESDDRRFNDPAWSQNPLYKRYLQTYLAWRKEL
                     HDWIGNSKLSEQDINRAHFVITLMTEAMAPTNSAANPAAVKRFFETGGKSLLDGLTHLA
                     KDLVNNGGMPSQVDMGAFEVGKSLGTTEGAVVFRNDVLELIQYRPTTEQVHERPLLVVP
                     PQINKFYVFDLSPDKSLARFCLSNNQQTFIVSWRNPTKAQREWGLSTYIDALKEAVDVV
                     SAITGSKDINMLGACSGGITCTALLGHYAALGEKKVNALTLLVTVLDTTLDSQVALFVD
                     EKTLEAAKRHSYQAGVLEGRDMAKVFAWMRPNDLIWNYWVNNYLLGNEPPVFDILFWNN
                     DTTRLPAAFHGDLIEMFKNNPLVRANALEVSGTPIDLKQVTADIYSLAGTNDHITPWKS
                     CYKSAQLFGGKVEFVLSSSGHIKSILNPPGNPKSRYMTSTDMPATANEWQENSTKHTDS
                     WWLHWQAWQAERSGKLKKSPTSLGNKAYPSGEAAPGTYVHER"
     gene            2616..4169
                     /gene="pct"
                     /label=pct
     CDS             2616..4169
                     /codon_start=1
                     /transl_table=11
                     /gene="pct"
                     /product="propionyl-CoA transferase"
                     /label=pct
                     /protein_id="QJR97784.1"
                     /translation="MRKVEIITAEQAAQLVKDNDTITSIGFVSSAHPEALTKALEKRFL
                     DTNTPQNLTYIYAGSQDKRDGRAAEHLAHTGLLKRAIIGHWQTVPAIGKLAVENKIEAY
                     NFSQGTLVHWFRALAGHKLGVFTDIGLETFLDPRQLGGKLNDVTKEDLVKLIEVDGHEQ
                     LFYPTFPVNVAFLRGTYADESGNITMDEEIGPFESTSVAQAVHNCGGKVVVQVKDVVAH
                     GSLDPRMVKIPGIYVDYVVVAAPEDHQQTYDCEYDPSLSGEHRAPEGATDAALPMSAKK
                     IIGRRGALELTENAVVNLGVGAPEYVASVAGEEGIADTITLTVEGGAIGGVPQGGARFG
                     SSRNADAIIDHTYQFDFYDGGGLDIAYLGLAQCDGSGNINVSKFGTNVAGCGGFPNISQ
                     QTPNVYFCGTFTAGGLKIAVEDGKVKILQEGKAKKFIKAVDQITFNGSYAARNGKHVLY
                     ITERCVFELTKEGLKLIEVAPGIDIEKDILAHMDFKPIIDNPKLMDARLFQDGPMGLKK
                     "
     terminator      4217..4248
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription
                     terminator"
     terminator      4373..4467
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      4959..5181
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             5229..6176
                     /label=Rep101
                     /note="RepA protein needed for replication with the pSC101 
                     origin"
     CDS             6468..7325
                     /label=AmpR
                     /note="beta-lactamase"
     terminator      7339..7358
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E.
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
ORIGIN
        1 ggtgtcacca ccctgccctt tttctttaaa accgaaaaga ttacttcgcg tttgccacct
       61 gacgtctaag aaaaggaata ttcagcaatt tgcccgtgcc gaagaaaggc ccacccgtga
      121 aggtgagcca gtgagttgat tgctacgtaa ttagttagtt agcccttagt gactcgaatt
      181 cttaagaccc actttcacat ttaagttgtt tttctaatcc gcatatgatc aattcaaggc
      241 cgaataagaa ggctggctct gcaccttggt gatcaaataa ttcgatagct tgtcgtaata
      301 atggcggcat actatcagta gtaggtgttt ccctttcttc tttagcgact tgatgctctt
      361 gatcttccaa tacgcaacct aaagtaaaat gccccacagc gctgagtgca tataatgcat
      421 tctctagtga aaaaccttgt tggcataaaa aggctaattg attttcgaga gtttcatact
      481 gtttttctgt aggccgtgta cctaaatgta cttttgctcc atcgcgatga cttagtaaag
      541 cacatctaaa acttttagcg ttattacgta aaaaatcttg ccagctttcc ccttctaaag
      601 ggcaaaagtg agtatggtgc ctatctaaca tctcaatggc taaggcgtcg agcaaagccc
      661 gcttattttt tacatgccaa tacaatgtag gctgctctac acctagcttc tgggcgagtt
      721 tacgggttgt taaaccttcg attccgacct cattaagcag ctctaatgcg ctgttaatca
      781 ctttactttt atctaatcta gacatcatta attcctaatt tttgttgaca ctctatcgtt
      841 gatagagtta ttttaccact ccctatcagt gatagagaaa aggtaccatt aaagaggaga
      901 aaacgcgtat gagcaacaaa aacagcgacg acctgaaccg tcaggcttcc gagaacacgc
      961 tgggtctgaa ccctgtaatc ggtctgcgtg gcaaggatct gctgacttct gcacgtatgg
     1021 tcctgactca agcaatcaag cagccaatcc actctgtcaa gcacgtcgca cacttcggta
     1081 tcgagctgaa gaacgtcatg ttcggtaaaa gcaaactgca accggagtcc gatgaccgtc
     1141 gtttcaacga cccggcatgg tctcagaacc ctctgtacaa acgttacctg caaacctacc
     1201 tggcctggcg taaagagctg cacgactgga tcggtaactc caaactgtcc gagcaggaca
     1261 tcaaccgtgc acacttcgtc atcacgctga tgaccgaagc tatggcacct actaactccg
     1321 ctgctaaccc agcagcagtg aaacgcttct tcgaaaccgg tggcaaaagc ctgctggacg
     1381 gtctgactca tctggcaaag gacctggtta acaacggtgg tatgccaagc caggttgaca
     1441 tgggtgcatt cgaagtgggc aaatctctgg gtactaccga aggtgcagtg gtgttccgta
     1501 acgacgtgct ggaactgatc cagtaccgtc caactactga acaggtgcac gaacgtccac
     1561 tgctggtagt gccgccgcaa attaacaaat tctacgtgtt cgacctgtcc ccggacaaat
     1621 ccctggctcg tttctgtctg tctaacaacc agcagacctt catcgtgtcc tggcgtaacc
     1681 cgactaaagc gcagcgtgaa tggggtctgt ctacttacat cgacgctctg aaagaagccg
     1741 ttgacgttgt ttccgccatc acgggtagca aagacatcaa catgctgggc gcttgctccg
     1801 gtggtattac ctgcactgct ctgctgggtc attacgctgc tctgggtgag aagaaagtga
     1861 acgctctgac tctgctggtt acggttctgg ataccaccct ggactctcag gttgctctgt
     1921 tcgttgatga gaaaacgctg gaagccgcta aacgccactc ttatcaggcg ggtgtactgg
     1981 aaggccgtga tatggcgaaa gttttcgcgt ggatgcgccc gaatgatctg atctggaact
     2041 actgggttaa caactacctg ctgggcaacg aaccgccggt ttttgatatc ctgttttgga
     2101 acaacgatac cacccgcctg ccggcggcgt ttcacggcga tctgattgaa atgtttaaaa
     2161 acaacccgct ggtacgcgcg aacgcgctgg aagtaagcgg caccccgatt gatctgaaac
     2221 aggttaccgc ggatatctat agcctggcgg gcaccaatga tcacattacc ccgtggaaat
     2281 cctgttacaa aagcgcccag ctgtttggcg gcaaagtaga atttgtactg agctccagcg
     2341 gccacattaa gagcattctg aatccgccgg gcaatccgaa atcccgctat atgacctcta
     2401 ccgatatgcc ggcgaccgcg aatgaatggc aggaaaattc taccaaacat accgattctt
     2461 ggtggctgca ttggcaggcg tggcaggccg aacgctctgg caaactgaag aaatctccga
     2521 cctctctggg caacaaagcc tatccgtctg gcgaagcggc gccgggcacc tatgttcatg
     2581 aacgctaaaa gctttcacac aggaaaggga tcctcatgag aaaagtagaa atcattactg
     2641 ctgaacaagc agctcagctc gtaaaagaca acgacacgat tacgtctatc ggctttgtca
     2701 gcagcgccca tccggaagca ctgaccaaag ctttggaaaa acggttcctg gacacgaaca
     2761 ccccgcagaa cttgacctac atctatgcag gctctcagga caaacgcgat ggccgtgccg
     2821 ctgaacatct ggcacacaca ggccttttga aacgcgccat catcggtcac tggcagactg
     2881 taccggctat cggtaaactg gctgtcgaaa acaagattga agcttacaac ttctcgcagg
     2941 gcacgttggt ccactggttc cgcgccttgg caggtcataa gctcggcgtc ttcaccgaca
     3001 tcggtctgga aactttcctc gatccccgtc agctcggcgg caagctcaat gacgtaacca
     3061 aagaagacct cgtcaaactg atcgaagtcg atggtcatga acagcttttc tacccgacct
     3121 tcccggtcaa cgtagctttc ctccgcggta cgtatgctga tgaatccggc aatatcacca
     3181 tggacgaaga aatcgggcct ttcgaaagca cttccgtagc ccaggccgtt cacaactgtg
     3241 gcggtaaagt cgtcgtccag gtcaaagacg tcgtcgctca cggcagcctc gacccgcgca
     3301 tggtcaagat ccctggcatc tatgtcgact acgtcgtcgt agcagctccg gaagaccatc
     3361 agcagacgta tgactgcgaa tacgatccgt ccctcagcgg tgaacatcgt gctcctgaag
     3421 gcgctaccga tgcagctctc cccatgagcg ctaagaaaat catcggccgc cgcggcgctt
     3481 tggaattgac tgaaaacgct gtcgtcaacc tcggcgtcgg tgctccggaa tacgttgctt
     3541 ctgttgccgg tgaagaaggt atcgccgata ccattaccct gaccgtcgaa ggtggcgcca
     3601 tcggtggcgt accgcagggc ggtgcccgct tcggttcgtc ccgcaatgcc gatgccatca
     3661 tcgaccacac ctatcagttc gacttctacg atggcggcgg tctggacatc gcttacctcg
     3721 gcctggccca gtgcgatggc tcgggcaaca tcaacgtcag caagttcggt actaacgttg
     3781 ccggctgcgg cggtttcccc aacatttccc agcagacacc gaatgtttac ttctgcggca
     3841 ccttcacggc tggcggcttg aaaatcgctg tcgaagacgg caaagtcaag atcctccagg
     3901 aaggcaaagc caagaagttc atcaaagctg tcgaccagat cactttcaac ggttcctatg
     3961 cagcccgcaa cggcaaacac gttctctaca tcacagaacg ctgcgtattt gaactgacca
     4021 aagaaggctt gaaactcatc gaagtcgcac cgggcatcga tattgaaaaa gatatcctcg
     4081 ctcacatgga cttcaagccg atcattgata atccgaaact catggatgcc cgcctcttcc
     4141 aggacggtcc catgggactg aaaaaataaa gatctctgca ggagtcacta agggttagtt
     4201 agttagatta gcagaaagtc aaaagcctcc gaccggaggc ttttgactaa aacttccctt
     4261 ggggttatca ttggggctca ctcaaaggcg gtaatcagat aaaaaaaatc cttagctttc
     4321 gctgaggatg atttctgcta gagctgtcag accaagttta cgagctcgct tggactcctg
     4381 ttgatagatc cagtaatgac ctcagaactc catctggatt tgttcagaac gctcggttgc
     4441 cgccgggcgt tttttattgg tgagaatcca agcactaggg acagtaagac gggtaagcct
     4501 gttgatgata ccgctgcctt actgggtgca ttagccagtc tgaatgacct gtcacggggt
     4561 aatccgaagt ggccagactg gaaaatcaga gggcaggaac tgctgaacag caaaaagtca
     4621 gatagcaccg catagcagac ccgccataaa acgccctgag aagcccgtga cgggcttttc
     4681 ttgtattatg ggtagtttcc ttgcatgaat ccataaaagg cgcctgtagt gccatttacc
     4741 cccattcact gccagagccg tgagtgcagc gaactgaatg tcacgaaaaa gacggcgact
     4801 caggtgcctg atggtcggag acaaaaggaa tattcagcga tttgcccgag cttgcgaggg
     4861 tgctacttaa gcctttaggg ttttaaggtc tgttttgtag aggagcaaac agcgtttgcg
     4921 acatcctttt gtaatactgc ggaactgact aaagtagtga gttatacaca gggctgggat
     4981 ctattctttt tatctttttt tattctttct ttattctata aattataacc acttgaatat
     5041 aaacaaaaaa aacacacaaa ggtctagcgg aatttacaga gggtctagca gaatttacaa
     5101 gttttccagc aaaggtctag cagaatttac agatacccac aactcaaagg aaaaggacat
     5161 gtaattatca ttgactagcc catctcaatt ggtatagtga ttaaaatcac ctagaccaat
     5221 tgagatgtat gtctgaatta gttgttttca aagcaaatga actagcgatt agtcgctatg
     5281 acttaacgga gcatgaaacc aagctaattt tatgctgtgt ggcactactc aaccccacga
     5341 ttgaaaaccc tacaaggaaa gaacggacgg tatcgttcac ttataaccaa tacgctcaga
     5401 tgatgaacat cagtagggaa aatgcttatg gtgtattagc taaagcaacc agagagctga
     5461 tgacgagaac tgtggaaatc aggaatcctt tggttaaagg ctttgagatt ttccagtgga
     5521 caaactgtgc caagttctca agcgaaaaat tagaattagt ttttagtgaa gggatattgc
     5581 cttatctttt ccagttaaaa aaattcataa aatataatct ggaacatgtt aagtcttttg
     5641 aaaacaaata ctctatgagg atttatgagt ggttattaaa agaactaaca caaaagaaaa
     5701 ctcacaaggc aaatatagag attagccttg atgaatttaa gttcatgtta atgcttgaaa
     5761 ataactacca tgagtttaaa aggcttaacc aatgggtttt gaaaccaata agtaaagatt
     5821 tgaacactta cagcaatatg aaattggtgg ttgataagcg aggccgcccg actgatacgt
     5881 tgattttcca agttgaacta gatagacaaa tggatctcgt aaccgaactt gagaacaacc
     5941 agataaaaat gaatggtgac aaaataccaa caaccattac atcagattcc tacctacata
     6001 acggactaag aaaaacacta cacgatgctt taactgcaaa aattcagttc accagttttg
     6061 aggcaaaatt tttgggtgac atgcaaagta agtatgatct caatggttcg ttctcatggc
     6121 tcacgcaaaa acaacgaacc acactagaga acatactggc taaatacgga aggatctgag
     6181 gttcttatgg ctcttgtatc tatcagtgaa gcatcaagac taacaaacaa aagtagaaca
     6241 actgttcacc gttacatatc aaagggaaaa ctgtccatat gcacagatga aaacggtgta
     6301 aaaaagatag atacatcaga gcttttacga gtttttggtg cattcaaagc tgttcaccat
     6361 gaacagatcg acaatgtaac tactagagct aaatacattc aaatatctat ccgctcatga
     6421 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagaatatg agtattcaac
     6481 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     6541 cagaaacgct ggtgaaagta aaagatgccg aagatcagtt gggcgcacgt gtgggttaca
     6601 tcgaactgga cctcaacagc ggtaagattc ttgagagttt tcgccccgaa gaacgtttcc
     6661 caatgatgag cacttttaaa gttctgctct gtggcgcggt attatcccgt attgacgccg
     6721 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     6781 cagtcacaga aaagcatctt acggacggca tgacagtacg cgaattatgc agcgctgcca
     6841 taaccacgag tgataacacg gcggccaact tacttctgac aacgatcggg ggaccgaagg
     6901 agcttaccgc ttttttgcac aacatgggtg atcatgtaac tcgccttgat cgttgggaac
     6961 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagctatgg
     7021 caacaacgtt gcgcaaactc ttaactggcg aacttcttac tctcgcttcc cggcaacaat
     7081 taatagactg gatggaggcg gataaagttg cgggaccact tctgcgctcg gcccttccgg
     7141 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtcccgc ggtattattg
     7201 cagccctggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagcc
     7261 aggcaactat ggacgaacgt aatcgccaga tcgctgagat aggtgcctcc ctgattaagc
     7321 attggtaata atactagctc cggcaaaaaa acgggcaa
//
