LOCUS       40924_48538        7416 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pZNR0943, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7416)
  AUTHORS   DeLoache WC, Russ ZN, Narcross L, Gonzales AM, Martin VJ, Dueber JE.
  TITLE     An enzyme-coupled biosensor enables (S)-reticuline production in 
            yeast from glucose
  JOURNAL   Nat. Chem. Biol. 11 (7), 465-471 (2015)
  PUBMED    25984720
REFERENCE   2  (bases 1 to 7416)
  AUTHORS   DeLoache WC, Russ ZN, Narcross L, Gonzales AM, Martin VJJ., Dueber 
            JE.
  TITLE     Direct Submission
  JOURNAL   Submitted (22-APR-2015) Department of Bioengineering, University of 
            California, Berkeley, 2151 Berkeley Way Room 542, Berkeley, CA 
            94704, USA
REFERENCE   3  (bases 1 to 7416)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7416)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat. Chem. 
            Biol."; date: "2015"; volume: "11"; issue: "7"; pages: "465-471"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (22-APR-2015) Department of Bioengineering, University of 
            California, Berkeley, 2151 Berkeley Way Room 542, Berkeley, CA 
            94704, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7416
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    12..17
                     /label=BsmBI recognition site
                     /note="BsmBI recognition site"
     misc_feature    19..22
                     /label=BsmBI recognition site
                     /note="BsmBI recognition site"
     regulatory      189..888
                     /regulatory_class="promoter"
     misc_feature    complement(894..897)
                     /label=BsaI recognition site
                     /note="BsaI recognition site"
     misc_feature    complement(899..904)
                     /label=BsaI recognition site
                     /note="BsaI recognition site"
     regulatory      905..1070
                     /regulatory_class="promoter"
     RBS             1053..1064
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1071..1784
                     /label=superfolder GFP
                     /note="GFP variant that folds robustly even when fused to
                     poorly folded proteins (Pedelacq et al., 2006)"
     misc_feature    1788..1793
                     /label=BsaI recognition site
                     /note="BsaI recognition site"
     misc_feature    1795..1798
                     /label=BsaI recognition site
                     /note="BsaI recognition site"
     CDS             1799..2491
                     /label=mKate2
                     /note="monomeric far-red fluorescent protein (Shcherbo et
                     al., 2009)"
     terminator      2501..2688
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae
                     alcohol dehydrogenase 1 (ADH1) gene"
     misc_feature    complement(2895..2898)
                     /label=BsmBI recognition site
                     /note="BsmBI recognition site"
     misc_feature    complement(2900..2905)
                     /label=BsmBI recognition site
                     /note="BsmBI recognition site"
     misc_feature    2917..3420
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an
                     autonomously replicating sequence"
     regulatory      complement(3426..3902)
                     /regulatory_class="terminator"
     CDS             complement(3906..4997)
                     /label=LEU2
                     /note="3-isopropylmalate dehydrogenase, required for
                     leucine biosynthesis"
     promoter        complement(4998..5405)
                     /label=LEU2 promoter
     rep_origin      complement(5761..6349)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     regulatory      6426..6573
                     /regulatory_class="promoter"
     CDS             6574..7386
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     regulatory      7390..7416
                     /regulatory_class="terminator"
ORIGIN
        1 gaattcgttc acgtctccgt tctggtagag ccacaaacag ccggtacaag caacgatctc
       61 caggaccatc tgaatcatgc gcggatgaca cgaactcacg acggcgatca cagacattaa
      121 cccacagccg ataattgcag acggtacaga cactgcgaca acgtggcaat tcgtcgcaat
      181 actctagaaa gaggatgtcc aatatttttt ttaaggaata aggatacttc aagactagat
      241 tcccccctgc attcccatca gaaccgtaaa ccttggcgct ttccttggga agtattcaag
      301 aagtgccttg tccggtttct gtggctcaca aaccagcgcg cccgatatgg ctttcttttc
      361 acttatgaat gtaccagtac gggacaatta gaacgctcct gtaacaatct ctttgcaaat
      421 gtggggttac attctaacca tgtcacactg ctgacgaaat tcaaagtaaa aaaaaatggg
      481 accacgtctt gagaacgata gattttcttt attttacatt gaacagtcgt tgtctcagcg
      541 cgctttatgt tttcattcat acttcatatt ataaaataac aaaagaagaa tttcatattc
      601 acgcccaaga aatcaggctg ctttccaaat gcaattgaca cttcattagc catcacacaa
      661 aactctttct tgctggagct tcttttaaaa aagacctcag tacaccaaac acgttacccg
      721 acctcgttat tttacgacaa ctatgataaa attctgaaga aaaaataaaa aaattttcat
      781 acttcttgct tttatttaaa ccattgaatg atttcttttg aacaaaacta cctgtttcac
      841 caaaggaaat agaaagaaaa aatcaattag aagaaaacaa aaaacaaaag atctatgtga
      901 gaccgaaagt gaaacgtgat ttcatgcgtc attttgaaca ttttgtaaat cttatttaat
      961 aatgtgtgcg gcaattcaca tttaatttat gaatgttttc ttaacatcgc ggcaactcaa
     1021 gaaacggcag gttcggatct tagctactag agaaagagga gaaatactag atgcgtaaag
     1081 gcgaagagct gttcactggt gtcgtcccta ttctggtgga actggatggt gatgtcaacg
     1141 gtcataagtt ttccgtgcgt ggcgagggtg aaggtgacgc aactaatggt aaactgacgc
     1201 tgaagttcat ctgtactact ggtaaactgc cggtaccttg gccgactctg gtaacgacgc
     1261 tgacttatgg tgttcagtgc tttgctcgtt atccggacca tatgaagcag catgacttct
     1321 tcaagtccgc catgccggaa ggctatgtgc aggaacgcac gatttccttt aaggatgacg
     1381 gcacgtacaa aacgcgtgcg gaagtgaaat ttgaaggcga taccctggta aaccgcattg
     1441 agctgaaagg cattgacttt aaagaagacg gcaatatcct gggccataag ctggaataca
     1501 attttaacag ccacaatgtt tacatcaccg ccgataaaca aaaaaatggc attaaagcga
     1561 attttaaaat tcgccacaac gtggaggatg gcagcgtgca gctggctgat cactaccagc
     1621 aaaacactcc aatcggtgat ggtcctgttc tgctgccaga caatcactat ctgagcacgc
     1681 aaagcgttct gtctaaagat ccgaacgaga aacgcgatca tatggttctg ctggagttcg
     1741 taaccgcagc gggcatcacg catggtatgg atgaactgta caaatgaggt ctccatccgt
     1801 gagcgagctg attaaggaga acatgcacat gaagctgtac atggagggca ccgtgaacaa
     1861 ccaccacttc aagtgcacat ccgagggcga aggcaagccc tacgagggca cccagaccat
     1921 gagaatcaag gcggtcgagg gcggccctct ccccttcgcc ttcgacatcc tggctaccag
     1981 cttcatgtac ggcagcaaaa ccttcatcaa ccacacccag ggcatccccg acttctttaa
     2041 gcagtccttc cccgagggct tcacatggga gagagtcacc acatacgaag acgggggcgt
     2101 gctgaccgct acccaggaca ccagcctcca ggacggctgc ctcatctaca acgtcaagat
     2161 cagaggggtg aacttcccat ccaacggccc tgtgatgcag aagaaaacac tcggctggga
     2221 ggcctccacc gaaaccctgt accccgctga cggcggcctg gaaggcagag ccgacatggc
     2281 cctgaagctc gttggcgggg gccacctgat ctgcaacttg aagaccacat acagatccaa
     2341 gaaacccgct aagaacctca agatgcccgg cgtctactat gtggacagaa gactggaaag
     2401 aatcaaggag gccgacaaag aaacctacgt cgagcagcac gaggtggctg tggccagata
     2461 ctgcgacctc cctagcaaac tggggcacag ataactcgag gcgaatttct tatgatttat
     2521 gatttttatt attaaataag ttataaaaaa aataagtgta tacaaatttt aaagtgactc
     2581 ttaggtttta aaacgaaaat tcttattctt gagtaactct ttcctgtagg tcaggttgct
     2641 ttctcaggta tagcatgagg tcgctcttat tgaccacacc tctaccggca tgccgagcaa
     2701 atgcctgcaa atcgctcccc atttcactag tgccacaagg tcagggcact catgcgacaa
     2761 tcaactcgat gcatgatccg caccattgtc gaggggccag caataaagct ccacacagtc
     2821 ggtcaatagt gccgaagacc acagacccgg ttaagacata gccgaatgga gccgcgccga
     2881 ccacagaatg atacacagag agacgcacag ctgcagatca cgtgctataa aaataattat
     2941 aatttaaatt ttttaatata aatatataaa ttaaaaatag aaagtaaaaa aagaaattaa
     3001 agaaaaaata gtttttgttt tccgaagatg taaaagactc tagggggatc gccaacaaat
     3061 actacctttt atcttgctct tcctgctctc aggtattaat gccgaattgt ttcatcttgt
     3121 ctgtgtagaa gaccacacac gaaaatcctg tgattttaca ttttacttat cgttaatcga
     3181 atgtatatct atttaatctg cttttcttgt ctaataaata tatatgtaaa gtacgctttt
     3241 tgttgaaatt ttttaaacct ttgtttattt ttttttcttc attccgtaac tcttctacct
     3301 tctttattta ctttctaaaa tccaaataca aaacataaaa ataaataaac acagagtaaa
     3361 ttcccaaatt attccatcat taaaagatac gaggcgcgtg taagttacag gcaagcgatc
     3421 cgtcatcgac tacgtcgtta aggccgtttc tgacagagta aaattcttga gggaactttc
     3481 accattatgg gaaatggttc aagaaggtat tgacttaaac tccatcaaat ggtcaggtca
     3541 ttgagtgttt tttatttgtt gtattttttt ttttttagag aaaatcctcc aatatataaa
     3601 ttaggaatca tagtttcatg attttctgtt acacctaact ttttgtgtgg tgccctcctc
     3661 cttgtcaata ttaatgttaa agtgcaattc tttttcctta tcacgttgag ccattagtat
     3721 caatttgctt acctgtattc ctttacatcc tcctttttct ccttcttgat aaatgtatgt
     3781 agattgcgta tatagtttcg tctaccctat gaacatattc cattttgtaa tttcgtgtcg
     3841 tttctattat gaatttcatt tataaagttt atgtacaaat atcataaaaa aagagaatct
     3901 ttttaagcaa ggattttctt aacttcttcg gcgacagcat caccgacttc ggtggtactg
     3961 ttggaaccac ctaaatcacc agttctgata cctgcatcca aaaccttttt aactgcatct
     4021 tcaatggcct taccttcttc aggcaagttc aatgacaatt tcaacatcat tgcagcagac
     4081 aagatagtgg cgatagggtt gaccttattc tttggcaaat ctggagcaga accgtggcat
     4141 ggttcgtaca aaccaaatgc ggtgttcttg tctggcaaag aggccaagga cgcagatggc
     4201 aacaaaccca aggaacctgg gataacggag gcttcatcgg agatgatatc accaaacatg
     4261 ttgctggtga ttataatacc atttaggtgg gttgggttct taactaggat catggcggca
     4321 gaatcaatca attgatgttg aaccttcaat gtaggaaatt cgttcttgat ggtttcctcc
     4381 acagtttttc tccataatct tgaagaggcc aaaacattag ctttatccaa ggaccaaata
     4441 ggcaatggtg gctcatgttg tagggccatg aaagcggcca ttcttgtgat tctttgcact
     4501 tctggaacgg tgtattgttc actatcccaa gcgacaccat caccatcgtc ttcctttctc
     4561 ttaccaaagt aaatacctcc cactaattct ctgacaacaa cgaagtcagt acctttagca
     4621 aattgtggct tgattggaga taagtctaaa agagagtcgg atgcaaagtt acatggtctt
     4681 aagttggcgt acaattgaag ttctttacgg atttttagta aaccttgttc aggtctaaca
     4741 ctaccggttc cccatttagg accacccaca gcacctaaca aaacggcatc agccttcttg
     4801 gaggcttcca gcgcctcatc tggaagtgga acacctgtag catcgatagc agcaccacca
     4861 attaaatgat tttcgaaatc gaacttgaca ttggaacgaa catcagaaat agctttaaga
     4921 accttaatgg cttcggctgt gatttcttga ccaacgtggt cacctggcaa aacgacgatc
     4981 ttcttagggg cagacattag aatggtatat ccttgaaata tatatatata tattgctgaa
     5041 atgtaaaagg taagaaaagt tagaaagtaa gacgattgct aaccacctat tggaaaaaac
     5101 aataggtcct taaataatat tgtcaacttc aagtattgtg atgcaagcat ttagtcatga
     5161 acgcttctct attctatatg aaaagccggt tccggcgctc tcacctttcc tttttctccc
     5221 aatttttcag ttgaaaaagg tatatgcgtc aggcgacctc tgaaattaac aaaaaatttc
     5281 cagtcatcga atttgattct gtgcgatagc gcccctgtgt gttctcgtta tgttgaggaa
     5341 aaaaataatg gttgctaaga gattcgaact cttgcatctt acgatacctg agtattccca
     5401 cagttaactg cggtcaagat atttcttgaa tcaggcgcct tagaccgctc ggccaaacaa
     5461 ccaattactt gttgagaaat agagtataat tatcctataa atataacgtt tttgaacaca
     5521 catgaacaag gaagtacagg acaattgatt ttgaagagaa tgtggatttt gatgtaattg
     5581 ttgggattcc atttttaata aggcaataat attaggtatg tagatatact agaagttctc
     5641 ctcgacggtc gggtaccccg aacggttatc cacagaatca ggggataacg caggaaagaa
     5701 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     5761 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     5821 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     5881 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     5941 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     6001 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     6061 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     6121 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     6181 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     6241 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     6301 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     6361 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     6421 catgactgat ccttcaactc agcaaaagtt cgatttattc aacaaagcca cgttgtgtct
     6481 caaaatctct gatgttacat tgcacaagat aaaaatatat catcatgaac aataaaactg
     6541 tctgcttaca taaacagtaa tacaaggggt gttatgagcc atattcaacg ggaaacgtct
     6601 tgctccaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta taaatgggct
     6661 cgcgataatg tcgggcaatc aggtgcgaca atctatcgat tgtatgggaa gcccgatgcg
     6721 ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac agatgagatg
     6781 gtcagactaa actggctgac ggaatttatg cctcttccga ccatcaagca ttttatccgt
     6841 actcctgatg atgcatggtt actcaccact gcgatccccg ggaaaacagc attccaggta
     6901 ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt gttcctgcgc
     6961 cggttgcatt cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt atttcgcctc
     7021 gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt tgatgacgag
     7081 cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataagctttt gccattctca
     7141 ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt tgacgagggg
     7201 aaattaatag gttgtattga tgttggacga gtcggaatcg cagaccgata ccaggatctt
     7261 gccatcctat ggaactgcct cggtgagttt tctccttcat tacagaaacg gctttttcaa
     7321 aaatatggta ttgataatcc tgatatgaat aaattgcagt ttcatttgat gctcgatgag
     7381 tttttctaat cagaattggt taattggttg taacac
//
