LOCUS       Exported                6080 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pTMH409, complete sequence.
ACCESSION   MF688771
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6080)
  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 6080)
  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 6080)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6080
                     /organism="Expression vector pTMH409"
                     /mol_type="other DNA"
                     /db_xref="taxon:2070487"
     CDS             complement(86..1168)
                     /codon_start=1
                     /product="LacI lac repressor"
                     /label=LacI lac repressor
                     /protein_id="AUO17142.1"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(1169..1246)
                     /gene="lacI"
                     /label=lacI promoter
     regulatory      1555..1574
                     /regulatory_class="promoter"
                     /note="T7"
     promoter        1555..1573
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1574..1598
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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)."
     protein_bind    1574..1596
                     /label=LacI binding site
                     /bound_moiety="LacI"
                     /note="LacO lac operator"
     RBS             1613..1635
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1642..3018
                     /codon_start=1
                     /product="flavin-containing monooxygenase FMO"
                     /EC_number="1.14.13.8"
                     /label=flavin-containing monooxygenase FMO
                     /note="from Methylophaga aminisulfidivorans"
                     /protein_id="AUO17145.1"
                     /translation="MATRIAILGAGPSGMAQLRAFQSAQEKGAEIPELVCFEKQADWGG
                     QWNYTWRTGLDENGEPVHSSMYRYLWSNGPKECLEFADYTFDEHFGKPIASYPPREVLW
                     DYIKGRVEKAGVRKYIRFNTAVRHVEFNEDSQTFTVTVQDHTTDTIYSEEFDYVVCCTG
                     HFSTPYVPEFEGFEKFGGRILHAHDFRDALEFKDKTVLLVGSSYSAEDIGSQCYKYGAK
                     KLISCYRTAPMGYKWPENWDERPNLVRVDTENAYFADGSSEKVDAIILCTGYIHHFPFL
                     NDDLRLVTNNRLWPLNLYKGVVWEDNPKFFYIGMQDQWYSFNMFDAQAWYARDVIMGRL
                     PLPSKEEMKADSMAWREKELTLVTAEEMYTYQGDYIQNLIDMTDYPSFDIPATNKTFLE
                     WKHHKKENIMTFRDHSYRSLMTGTMAPKHHTPWIDALDDSLEAYLSDKSEIPVAKEAGS
                     "
     regulatory      3027..3038
                     /regulatory_class="ribosome_binding_site"
     RBS             3027..3038
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             3045..4478
                     /codon_start=1
                     /product="PtUGT72B29 indoxyl glucosyltransferase"
                     /label=PtUGT72B29 indoxyl glucosyltransferase
                     /note="from Polygonum tinctorium"
                     /protein_id="AUO17143.1"
                     /translation="MESPAAPPTTAPPPHVIIVPSAGMGHLIPLAEFAKRLLPRFTFTF
                     AVPTSGPPSSSQRDFLSSLPASIDTSFLPEVDLSDAPSDAQIETLMSLMVVRSLPSLRD
                     LIASYSASGRRVAALVVDLFATDAIDVALELGIRPFIFFPSTAMTLSFFLHLEKLDETV
                     SCEFAELSDPVQIPGCIPVHGKDLIDPVQDRKNDAYKWLLHHSKRYKLAEGVIVNSFEG
                     LEGGPIRELLHPEPGKPRVYPVGPLIQAGSCEKGAAARPECLKWLDQQPRGSVLFVNFG
                     SGGVLSTEQQNELAGVLAHSQQRFLWVVRPPNDGIANATYFSVDGEIDPLKLLPEGFLE
                     QTAGRGLVLPMWAPQIDVLSHESTGGFLTHCGWNSTLESVFHGVPLITWPLYAEQKMNA
                     VMLTEGLRVGLRPSVGKDGIIRGAEIARVIGELMEGEEGKRIRSKMQELKRAASAVLSK
                     DGSSTRALEEVAKIWESKV"
     regulatory      4556..4603
                     /regulatory_class="terminator"
                     /note="T7 terminator"
     terminator      4556..4603
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     regulatory      complement(4659..4685)
                     /regulatory_class="terminator"
     CDS             complement(4686..5501)
                     /codon_start=1
                     /product="kanamycin resistance protein"
                     /label=kanamycin resistance protein
                     /protein_id="AUO17144.1"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     regulatory      complement(5502..5649)
                     /regulatory_class="promoter"
     rep_origin      5650..6049
                     /label=R6K origin
                     /note="R6K origin"
     rep_origin      5666..6022
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"