LOCUS       40924_43613        7150 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pTMH308, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7150)
  AUTHORS   Hsu TM, Welner DH, Russ ZN, Cervantes B, Prathuri RL, Adams PD, 
            Dueber JE.
  TITLE     Employing a biochemical protecting group for a sustainable indigo 
            dyeing strategy
  JOURNAL   Nat. Chem. Biol. (2018) In press
  PUBMED    29309053
REFERENCE   2  (bases 1 to 7150)
  AUTHORS   Hsu TM, Welner DH, Russ ZN, Cervantes B, Prathuri RL, Adams PD, 
            Dueber JE.
  TITLE     Direct Submission
  JOURNAL   Submitted (20-AUG-2017) Bioengineering, UC Berkeley, 2151 Berkeley 
            Way Room 512D, Berkeley, CA 94704, USA
REFERENCE   3  (bases 1 to 7150)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7150)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat. Chem. 
            Biol. (2018) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (20-AUG-2017) Bioengineering, UC Berkeley, 2151 Berkeley Way Room 
            512D, Berkeley, CA 94704, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7150
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    complement(168..310)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(415..603)
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     protein_bind    complement(1881..1902)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(1918..2997)
                     /label=lacI
                     /note="lac repressor"
     promoter        complement(2998..3075)
                     /label=lacI promoter
     promoter        3384..3402
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3403..3427
                     /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             3442..3464
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3477..3494
                     /label=6xHis
                     /note="6xHis affinity tag"
     CDS             3516..3536
                     /label=TEV site
                     /note="tobacco etch virus (TEV) protease recognition and 
                     cleavage site"
     misc_feature    3540..4967
                     /note="PtUGT72B30 indoxyl glucosyltransferase from
                     Polygonum tinctorium"
     terminator      5045..5092
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(5267..5295)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein
                     gene"
     promoter        5408..5512
                     /label=AmpR promoter
     CDS             5513..6370
                     /label=AmpR
                     /note="beta-lactamase"
     regulatory      6371..6467
                     /regulatory_class="terminator"
     rep_origin      6544..7132
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
       61 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
      121 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat
      181 tttctcctta cgcatctgtg cggtatttca caccgcatat atggtgcact ctcagtacaa
      241 tctgctctga tgccgcatag ttaagccagt atacactccg ctatcgctac gtgactgggt
      301 catggctgcg ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
      361 cccggcatcc gcttacagac aagctgtgac cttctccggg agctgcatgt gtcagaggtt
      421 ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg
      481 aagcgattca cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt tctccagaag
      541 cgttaatgtc tggcttctga taaagcgggc catgttaagg gcggtttttt cctgtttggt
      601 cactgatgcc tccgtgtaag ggggatttct gttcatgggg gtaatgatac cgatgaaacg
      661 agagaggatg ctcacgatac gggttactga tgatgaacat gcccggttac tggaacgttg
      721 tgagggtaaa caactggcgg tatggatgcg gcgggaccag agaaaaatca ctcagggtca
      781 atgccagcgc ttcgttaata cagatgtagg tgttccacag ggtagccagc agcatcctgc
      841 gatgcagatc cggaacataa tggtgcaggg cgctgacttc cgcgtttcca gactttacga
      901 aacacggaaa ccgaagacca ttcatgttgt tgctcaggtc gcagacgttt tgcagcagca
      961 gtcgcttcac gttcgctcgc gtatcggtga ttcattctgc taaccagtaa ggcaaccccg
     1021 ccagcctagc cgggtcctca acgacaggag cacgatcatg cgcacccgtg gccaggaccc
     1081 aacgctgccc gagatgcgcc gcgtgcggct gctggagatg gcggacgcga tggatatgtt
     1141 ctgccaaggg ttggtttgcg cattcacagt tctccgcaag aattgattgg ctccaattct
     1201 tggagtggtg aatccgttag cgaggtgccg ccggcttcca ttcaggtcga ggtggcccgg
     1261 ctccatgcac cgcgacgcaa cgcggggagg cagacaaggt atagggcggc gcctacaatc
     1321 catgccaacc cgttccatgt gctcgccgag gcggcataaa tcgccgtgac gatcagcggt
     1381 ccagtgatcg aagttaggct ggtaagagcc gcgagcgatc cttgaagctg tccctgatgg
     1441 tcgtcatcta cctgcctgga cagcatggcc tgcaacgcgg gcatcccgat gccgccggaa
     1501 gcgagaagaa tcataatggg gaaggccatc cagcctcgcg tcgcgaacgc cagcaagacg
     1561 tagcccagcg cgtcggccgc catgccggcg ataatggcct gcttctcgcc gaaacgtttg
     1621 gtggcgggac cagtgacgaa ggcttgagcg agggcgtgca agattccgaa taccgcaagc
     1681 gacaggccga tcatcgtcgc gctccagcga aagcggtcct cgccgaaaat gacccagagc
     1741 gctgccggca cctgtcctac gagttgcatg ataaagaaga cagtcataag tgcggcgacg
     1801 atagtcatgc cccgcgccca ccggaaggag ctgactgggt tgaaggctct caagggcatc
     1861 ggtcgagatc ccggtgccta atgagtgagc taacttacat taattgcgtt gcgctcactg
     1921 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     1981 gggagaggcg gtttgcgtat tgggcgccag ggtggttttt cttttcacca gtgaaacggg
     2041 caacagctga ttgcccttca ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct
     2101 ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt aacggcggga tataacatga
     2161 gctgtcttcg gtatcgtcgt atcccactac cgagatatcc gcaccaacgc gcagcccgga
     2221 ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg ttggcaacca gcatcgcagt
     2281 gggaacgatg ccctcattca gcatttgcat ggtttgttga aaaccggaca tggcactcca
     2341 gtcgccttcc cgttccgcta tcggctgaat ttgattgcga gtgagatatt tatgccagcc
     2401 agccagacgc agacgcgccg agacagaact taatgggccc gctaacagcg cgatttgctg
     2461 gtgacccaat gcgaccagat gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat
     2521 aatactgttg atgggtgtct ggtcagagac atcaagaaat aacgccggaa cattagtgca
     2581 ggcagcttcc acagcaatgg catcctggtc atccagcgga tagttaatga tcagcccact
     2641 gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc
     2701 taccatcgac accaccacgc tggcacccag ttgatcggcg cgagatttaa tcgccgcgac
     2761 aatttgcgac ggcgcgtgca gggccagact ggaggtggca acgccaatca gcaacgactg
     2821 tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc
     2881 ttccactttt tcccgcgttt tcgcagaaac gtggctggcc tggttcacca cgcgggaaac
     2941 ggtctgataa gagacaccgg catactctgc gacatcgtat aacgttactg gtttcacatt
     3001 caccaccctg aattgactct cttccgggcg ctatcatgcc ataccgcgaa aggttttgcg
     3061 ccattcgatg gtgtccggga tctcgacgct ctcccttatg cgactcctgc attaggaagc
     3121 agcccagtag taggttgagg ccgttgagca ccgccgccgc aaggaatggt gcatgcaagg
     3181 agatggcgcc caacagtccc ccggccacgg ggcctgccac catacccacg ccgaaacaag
     3241 cgctcatgag cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg gcgatatagg
     3301 cgccagcaac cgcacctgtg gcgccggtga tgccggccac gatgcgtccg gcgtagagga
     3361 tcgagatctc gatcccgcga aattaatacg actcactata ggggaattgt gagcggataa
     3421 caattcccct ctagaaataa ttttgtttaa ctttaagaag gagatatacc atgggccatc
     3481 accatcacca tcacgactac gacatcccga ctaccgaaaa cctgtacttc cagggttctg
     3541 aatcccccgc cgctccacca accaccgctc caccaccgca cgtcataatc atgccctccg
     3601 ccggcatggg ccacctcatc cccctcgccg agttcgccaa gcgcctcctt ccgcgcttca
     3661 ccttcacctt cgccgtaccc accagcggcc cgccctcatc ctcccagcgc gacttcctct
     3721 cctccctccc tgcctccatc gacacctcct tcctccccga ggtcgacctc tccgacgccc
     3781 cctccgacgc ccaaatcgag actctcatgt ccctcatggt tgtccgctcc ctcccctcgc
     3841 tccgcgacct cattgcctcc tactccgcct ccggccgccg cgtcgccgcc ctcgtcgtcg
     3901 accttttcgc cactgatgca atcgacgtcg cccttgagct cggcatccgc cctttcatct
     3961 tcttcccctc caccgccatg accctctcct tcttcctcca cctcgagaag cttgatgaaa
     4021 cggtgtcatg tgagtttgcc gagctgtccg acccggttca gatccccggg tgtattccgg
     4081 tccacggcaa ggatttgatc gacccggttc aggataggaa gaacgacgcc tacaagtggc
     4141 tcctccacca ctccaagagg tataaattgg ccgagggtgt tatcgtaaat agcttcgagg
     4201 gtttggaggc cggaccgatc aggcagcttt tgcaccccga gcctggcaag ccgcgggttt
     4261 acccggtcgg accgctgatt caggccggtt cgtgcgagaa gggggcagct gcccggcctg
     4321 agtgcttgaa gtggttggac cagcagccac gtggatccgt cctattcgtg aatttcggga
     4381 gtggtggggt cctcagtacg gagcagcaga acgagcttgc aggtgtgctg gcccacagcc
     4441 agcagaggtt cctatgggtg gttaggcctc caaacgacgg cattgccaac gccacgtatt
     4501 tcagcgtcga cggggagatc gacccgttga aactcctgcc cgaggggttc ttggagcaga
     4561 ccgcgggcag gggtttggtt ttgccaatgt gggccccgca gatcgatgtc ttaagccatg
     4621 agtcgacggg cgggttcttg acgcattgtg ggtggaattc aacactagag agcgtgttcc
     4681 atggggtacc actaattaca tggcccctct atgccgagca aaagatgaac gctgttatgc
     4741 tgaccgaggg cctgagggtg ggactcagac cctcagtggg taaggatgga atcatccgag
     4801 gtgatgagat cgcacgagtt ataggggagt tgatggaagg cgaggaaggg aaacgaatac
     4861 ggagtaagat gcaggagttg aagcgtgcgg cttctgctgt attgagcaaa gatggatcgt
     4921 ctactcgagc tcttgaagag gttgcaaaaa tttgggaaag caaggtttaa ggatcctgat
     4981 caatccggct gctaacaaag cccgaaagga agctgagttg gctgctgcca ccgctgagca
     5041 ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt tgctgaaagg
     5101 aggaactata tccggatatc ccgcaagagg cccggcagta ccggcataac caagcctatg
     5161 cctacagcat ccagggtgac ggtgccgagg atgacgatga gcgcattgtt agatttcata
     5221 cacggtgcct gactgcgtta gcaatttaac tgtgataaac taccgcatta aagcttatcg
     5281 atgataagct gtcaaacatg agaattcttg aagacgaaag ggcctcgtga tacgcctatt
     5341 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
     5401 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     5461 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     5521 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     5581 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     5641 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     5701 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtgttga
     5761 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     5821 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     5881 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     5941 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     6001 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgcagc
     6061 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     6121 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     6181 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt cacgcggtat
     6241 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     6301 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     6361 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     6421 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     6481 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     6541 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     6601 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     6661 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca
     6721 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     6781 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     6841 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     6901 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     6961 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     7021 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     7081 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     7141 caacgcggcc
//
