LOCUS       40924_31085        5540 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pML1381, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5540)
  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 5540)
  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 5540)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5540)
  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..5540
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    5..524
                     /label=similar to LEU2 5'
                     /note="similar to LEU2 5'"
     misc_feature    687..692
                     /label=BsmBI recognition site
                     /note="BsmBI recognition site"
     misc_feature    694..697
                     /label=BsmBI recognition site
                     /note="BsmBI recognition site"
     misc_feature    complement(764..767)
                     /label=BsmBI recognition site
                     /note="BsmBI recognition site"
     misc_feature    complement(769..774)
                     /label=BsmBI recognition site
                     /note="BsmBI recognition site"
     promoter        1177..1584
                     /label=LEU2 promoter
     CDS             1585..2676
                     /label=LEU2
                     /note="3-isopropylmalate dehydrogenase, required for
                     leucine biosynthesis"
     regulatory      2680..3156
                     /regulatory_class="terminator"
     misc_feature    3161..3670
                     /label=similar to LEU2 3'
                     /note="similar to LEU2 3'"
     regulatory      complement(3683..3779)
                     /regulatory_class="terminator"
     CDS             complement(3780..4637)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(4638..4742)
                     /label=AmpR promoter
     rep_origin      4845..5433
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 caatacccag aagaacagta aaataaagca aggtacgtga aattaatatt tttaaatggt
       61 tctaaccgat gccgaagaac tgcgcagtcc ggttataacg tctgacatgt ccttttttga
      121 tttggaatcc aaccactcaa gtgactctgt tcatttactt tgcgaaaaat atacccacaa
      181 attgcccatc gaaagtgaat cgcaaaccac cttcagactg gcaccgacaa agcaaagatt
      241 atacagacag agtactttat acgtaccgtt aagtctcaag caaagggttt tcttatttac
      301 tgaacgggta aagagtatct gggccggctt gccaagatgc aaaccgaata agtatttcaa
      361 agttgcattt gccttagccg tcctgacacc attggctatt tggatatttt atattgactt
      421 tcgtgtacat tgatcacatc gactgttcta ttggcaaatg aaccacgggc attgactatt
      481 tttcaggtta ctactatata ttatcatcac gggcaaggat tgtaccctga attcgcatct
      541 agaacggggt catcacggct catcatgcgc caaacaaatg tgtgcaatac acgctcggat
      601 gactgcatga tgaccgcact gactggggac agcagatcca cctaagcctg tgagagaagc
      661 agacacccga cagatcaagg cagttacgtc tcaccaaacc agatgtcaac acagctacaa
      721 cgactagaga tctatgtgag gatcctaact cgagatcgag ctgagcatga gacggaaatc
      781 tgctcgtcag tggtgctcac actgacgaat catgtacaga tcataccgat gactgcctgg
      841 cgactcacaa ctaagcaaga cagccggaac cagcgccggc gaacaccact gcatatatgg
      901 catatcacaa cagtccaact agtgcactgc agtacatcga ggagaacttc tagtatatct
      961 acatacctaa tattattgcc ttattaaaaa tggaatccca acaattacat caaaatccac
     1021 attctcttca aaatcaattg tcctgtactt ccttgttcat gtgtgttcaa aaacgttata
     1081 tttataggat aattatactc tatttctcaa caagtaattg gttgtttggc cgagcggtct
     1141 aaggcgcctg attcaagaaa tatcttgacc gcagttaact gtgggaatac tcaggtatcg
     1201 taagatgcaa gagttcgaat ctcttagcaa ccattatttt tttcctcaac ataacgagaa
     1261 cacacagggg cgctatcgca cagaatcaaa ttcgatgact ggaaattttt tgttaatttc
     1321 agaggtcgcc tgacgcatat acctttttca actgaaaaat tgggagaaaa aggaaaggtg
     1381 agagcgccgg aaccggcttt tcatatagaa tagagaagcg ttcatgacta aatgcttgca
     1441 tcacaatact tgaagttgac aatattattt aaggacctat tgttttttcc aataggtggt
     1501 tagcaatcgt cttactttct aacttttctt accttttaca tttcagcaat atatatatat
     1561 atatttcaag gatataccat tctaatgtct gcccctaaga agatcgtcgt tttgccaggt
     1621 gaccacgttg gtcaagaaat cacagccgaa gccattaagg ttcttaaagc tatttctgat
     1681 gttcgttcca atgtcaagtt cgatttcgaa aatcatttaa ttggtggtgc tgctatcgat
     1741 gctacaggtg ttccacttcc agatgaggcg ctggaagcct ccaagaaggc tgatgccgtt
     1801 ttgttaggtg ctgtgggtgg tcctaaatgg ggaaccggta gtgttagacc tgaacaaggt
     1861 ttactaaaaa tccgtaaaga acttcaattg tacgccaact taagaccatg taactttgca
     1921 tccgactctc ttttagactt atctccaatc aagccacaat ttgctaaagg tactgacttc
     1981 gttgttgtca gagaattagt gggaggtatt tactttggta agagaaagga agacgatggt
     2041 gatggtgtcg cttgggatag tgaacaatac accgttccag aagtgcaaag aatcacaaga
     2101 atggccgctt tcatggccct acaacatgag ccaccattgc ctatttggtc cttggataaa
     2161 gctaatgttt tggcctcttc aagattatgg agaaaaactg tggaggaaac catcaagaac
     2221 gaatttccta cattgaaggt tcaacatcaa ttgattgatt ctgccgccat gatcctagtt
     2281 aagaacccaa cccacctaaa tggtattata atcaccagca acatgtttgg tgatatcatc
     2341 tccgatgaag cctccgttat cccaggttcc ttgggtttgt tgccatctgc gtccttggcc
     2401 tctttgccag acaagaacac cgcatttggt ttgtacgaac catgccacgg ttctgctcca
     2461 gatttgccaa agaataaggt caaccctatc gccactatct tgtctgctgc aatgatgttg
     2521 aaattgtcat tgaacttgcc tgaagaaggt aaggccattg aagatgcagt taaaaaggtt
     2581 ttggatgcag gtatcagaac tggtgattta ggtggttcca acagtaccac cgaagtcggt
     2641 gatgctgtcg ccgaagaagt taagaaaatc cttgcttaaa aagattctct ttttttatga
     2701 tatttgtaca taaactttat aaatgaaatt cataatagaa acgacacgaa attacaaaat
     2761 ggaatatgtt catagggtag acgaaactat atacgcaatc tacatacatt tatcaagaag
     2821 gagaaaaagg aggatgtaaa ggaatacagg taagcaaatt gatactaatg gctcaacgtg
     2881 ataaggaaaa agaattgcac tttaacatta atattgacaa ggaggagggc accacacaaa
     2941 aagttaggtg taacagaaaa tcatgaaact atgattccta atttatatat tggaggattt
     3001 tctctaaaaa aaaaaaaata caacaaataa aaaacactca atgacctgac catttgatgg
     3061 agtttaagtc aataccttct tgaaccattt cccataatgg tgaaagttcc ctcaagaatt
     3121 ttactctgtc agaaacggcc ttaacgacgt agtcgagagt actcgtatcg catgtcggtg
     3181 cgacacgaaa ttacaaaatg gaatatgttc atagggtaga cgaaactata tacgcaatct
     3241 acatacattt atcaagaagg agaaaaagga ggatgtaaag gaatacaggt aagcaaattg
     3301 atactaatgg ctcaacgtga taaggaaaaa gaattgcact ttaacattaa tattgacaag
     3361 gaggagggca ccacacaaaa agttaggtgt aacagaaaat catgaaacta tgattcctaa
     3421 tttatatatt ggaggatttt ctctaaaaaa aaaaaaatac aacaaataaa aaacactcaa
     3481 tgacctgacc atttgatgga gtttaagtca ataccttctt gaaccatttc ccataatggt
     3541 gaaagttccc tcaagaattt tactctgtca gaaacggcct taacgacgta gtcgacctcc
     3601 tcttcagtac taaatctacc aataccaaat ctgatggaag aatgggctaa tgcatcatcc
     3661 ttacccagcg ccgagcggcc gcgattatca aaaaggatct tcacctagat ccttttaaat
     3721 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     3781 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     3841 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     3901 gctgcaatga taccgcggga cccacgctca ccggctccag atttatcagc aataaaccag
     3961 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     4021 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     4081 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     4141 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     4201 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     4261 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     4321 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     4381 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     4441 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     4501 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     4561 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     4621 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     4681 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     4741 cgcacatttc cccgaaaagt gccacctgtc atgaccaaaa tcccttaacg tgagttttcg
     4801 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     4861 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     4921 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     4981 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     5041 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     5101 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     5161 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     5221 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     5281 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     5341 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     5401 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     5461 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     5521 gtggataacc gtgcggccgc
//
