LOCUS       sequence-422586-        7054 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TtgV Biosensor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7054)
  AUTHORS   Pham C, Stogios PJ, Savchenko A, Mahadevan R
  TITLE     Design and Characterization of a Generalist Biosensor for Indole 
            Derivatives.
  JOURNAL   ACS Synth Biol. 2024 Jul 19;13(7):2246-2252. doi: 
            10.1021/acssynbio.3c00736. Epub 2024 Jun 14.
  PUBMED    38875315
REFERENCE   2  (bases 1 to 7054)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1021/acssynbio.3c00736"; journalName: "ACS Synth Biol"; date: 
            "2024-07-19- 19"; volume: "13"; issue: "7"; pages: "2246-2252"
COMMENT     Sequence Label: pST
FEATURES             Location/Qualifiers
     source          1..7054
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             112..1194
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEATCTNVPALFLDVSDQTPINSIIFSH
                     EDGMRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(131..150)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             1213..1836
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     complement(1286..1305)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     rep_origin      complement(2403..2948)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(2483..2502)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3037..3852)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(3037..3852)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     3760..3779
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3844..3863
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     promoter        join(3974..3979,3980..3995,3996..4002)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    4008..4027
                     /label=lac operator (symmetric)
                     /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). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
     RBS             5076..5084
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             5090..5803
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             5834..7024
                     /codon_start=1
                     /gene="tet"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSIIGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
ORIGIN
        1 ggggggcctt ttttcgtttt ggtccaatgg cggcgcgcca tcgaatggcg caaaaccttt
       61 cgcggtatgg catgatagcg cccggaagag agtcaattca gggtggtgaa tatgaaacca
      121 gtaacgttat acgatgtcgc agagtatgcc ggtgtctctt atcagaccgt ttcccgcgtg
      181 gtgaaccagg ccagccacgt ttctgcgaaa acgcgggaaa aagtggaagc ggcgatggcg
      241 gagctgaatt acattcccaa ccgcgtggca caacaactgg cgggcaaaca gtcgttgctg
      301 attggcgttg ccacctccag tctggccctg cacgcgccgt cgcaaattgt cgcggcgatt
      361 aaatctcgcg ccgatcaact gggtgccagc gtggtggtgt cgatggtaga acgaagcggc
      421 gtcgaagcct gtaaagcggc ggtgcacaat cttctcgcgc aacgcgtcag tgggctgatc
      481 attaactatc cgctggatga ccaggatgcc attgctgtgg aagctacctg cactaatgtt
      541 ccggcgttat ttcttgatgt ctctgaccag acacccatca acagtattat tttctcccat
      601 gaggacggta tgcgactggg cgtggagcat ctggtcgcat tgggtcacca gcaaatcgcg
      661 ctgttagcgg gcccattaag ttctgtctcg gcgcgtctgc gtctggctgg ctggcataaa
      721 tatctcactc gcaatcaaat tcagccgata gcggaacggg aaggcgactg gagtgccatg
      781 tccggttttc aacaaaccat gcaaatgctg aatgagggca tcgttcccac tgcgatgctg
      841 gttgccaacg atcagatggc gctgggcgca atgcgcgcca ttaccgagtc cgggctgcgc
      901 gttggtgcgg atatctcggt agtgggatac gacgataccg aagatagctc atgttatatc
      961 ccgccgttaa ccaccatcaa acaggatttt cgcctgctgg ggcaaaccag cgtggaccgc
     1021 ttgctgcaac tctctcaggg ccaggcggtg aagggcaatc agctgttgcc agtctcactg
     1081 gtgaaaagaa aaaccaccct ggcgcccaat acgcaaaccg cctccccccg cgcgttggcc
     1141 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgataatcc
     1201 aggaggaaaa aaatgtccag attagataaa agtaaagtga ttaacagcgc attagagctg
     1261 cttaatgagg tcggaatcga aggtttaaca acccgtaaac tcgcccagaa gctaggtgta
     1321 gagcagccta cattgtattg gcatgtaaaa aataagcggg ctttgctcga cgccttagcc
     1381 attgagatgt tagataggca ccatactcac ttttgccctt tagaagggga aagctggcaa
     1441 gattttttac gtaataacgc taaaagtttt agatgtgctt tactaagtca tcgcgatgga
     1501 gcaaaagtac atttaggtac acggcctaca gaaaaacagt atgaaactct cgaaaatcaa
     1561 ttagcctttt tatgccaaca aggtttttca ctagagaatg cattatatgc actcagcgct
     1621 gtggggcatt ttactttagg ttgcgtattg gaagatcaag agcatcaagt cgctaaagaa
     1681 gaaagggaaa cacctactac tgatagtatg ccgccattat tacgacaagc tatcgaatta
     1741 tttgatcacc aaggtgcaga gccagccttc ttattcggcc ttgaattgat catatgcgga
     1801 ttagaaaaac aacttaaatg tgaaagtggg tcctaataat tggtaacgaa tcagacaatt
     1861 gacggctcga gggagtagca tagggtttgc agaatccctg cttcgtccat ttgacaggca
     1921 cattatgcat cgatgataag ctgtcaaaca tgagcagatc ctctacgccg gacgcatcgt
     1981 ggccggcatc accggcgcca caggtgcggt tgctggcgcc tatatcgccg acatcaccga
     2041 tggggaagat cgggctcgcc acttcgggct catgagcaaa tgttttatct gaggtgcttc
     2101 ctcgctcact gactcgctgc acgaggcaga cctcagcgct agcggagtgt atactggctt
     2161 actatgttgg cactgatgag ggtgtcagtg aagtgcttca tgtggcagga gaaaaaaggc
     2221 tgcaccggtg cgtcagcaga atatgtgata caggatatat tccgcttcct cgctcactga
     2281 ctcgctacgc tcggtcgttc gactgcggcg agcggaaatg gcttacgaac ggggcggaga
     2341 tttcctggaa gatgccagga agatacttaa cagggaagtg agagggccgc ggcaaagccg
     2401 tttttccata ggctccgccc ccctgacaag catcacgaaa tctgacgctc aaatcagtgg
     2461 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggcgg ctccctcgtg
     2521 cgctctcctg ttcctgcctt tcggtttacc ggtgtcattc cgctgttatg gccgcgtttg
     2581 tctcattcca cgcctgacac tcagttccgg gtaggcagtt cgctccaagc tggactgtat
     2641 gcacgaaccc cccgttcagt ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     2701 caacccggaa agacatgcaa aagcaccact ggcagcagcc actggtaatt gatttagagg
     2761 agttagtctt gaagtcatgc gccggttaag gctaaactga aaggacaagt tttggtgact
     2821 gcgctcctcc aagccagtta cctcggttca aagagttggt agctcagaga accttcgaaa
     2881 aaccgccctg caaggcggtt ttttcgtttt cagagcaaga gattacgcgc agaccaaaac
     2941 gatctcaaga agatcatctt attaaggggt ctgacgctca gtggaacgaa aaatcaatct
     3001 aaagtatata tgagtaaact tggtctgaca gttaccttag aaaaactcat cgagcatcaa
     3061 atgaaactgc aatttattca tatcaggatt atcaatacca tatttttgaa aaagccgttt
     3121 ctgtaatgaa ggagaaaact caccgaggca gttccatagg atggcaagat cctggtatcg
     3181 gtctgcgatt ccgactcgtc caacatcaat acaacctatt aatttcccct cgtcaaaaat
     3241 aaggttatca agtgagaaat caccatgagt gacgactgaa tccggtgaga atggcaaaag
     3301 cttatgcatt tctttccaga cttgttcaac aggccagcca ttacgctcgt catcaaaatc
     3361 actcgcatca accaaaccgt tattcattcg tgattgcgcc tgagcgagac gaaatacgcg
     3421 atcgctatta aaaggacaat tacaaacagg aatcgaatgc aaccggcgca ggaacactgc
     3481 cagcgcatca acaatgtttt cacctgaatc aggatattct tctaatacct ggaatgctgt
     3541 tttcccgggg atcgcagtgg tgagtaacca tgcatcatca ggagtacgga taaaatgctt
     3601 gatggtcgga agaggcataa attccgtcag ccagtttagt ctgaccatct catctgtaac
     3661 atcattggca acgctacctt tgccatgttt cagaaacaac tctggcgcat cgggcttccc
     3721 atacaatcga tagattgtcg cacctgattg cccgacatta tcgcgagccc atttataccc
     3781 atataaatca gcatccatgt tggaatttaa tcgcggcctg gagcaagacg tttcccgttg
     3841 aatatggctc ataacacccc ttgtattacc gtttatgtaa gcagacagtt ttattgttca
     3901 tgatgatata tttttatctt gtgcaatgta catcagagat tttgagacac aaggagaacg
     3961 atcgttggct gtgttgacaa ttaatcatcg gctcgtataa tgtgtggaat tgtgagcgct
     4021 cacaattagt cgtcaagtgc tgtgcttgca cttctgatga ggcagtgatg ccgaaacgac
     4081 ctctacaaat aattttgttt aagccatgtc attggctttt gatagaggac aactactaga
     4141 tgaatcaatc ggacgagaac atcggtaaag cgggtggtat tcaagttatt gcacgtgcgg
     4201 cgtctattat gcgtgccctc ggttcccacc cccatggtct ttctctggcc gcaatcgccc
     4261 agcttgtggg tttgccgcgt agtacggtgc aacgtattat taatgcgtta gaagaggaat
     4321 ttcttgtgga agccctgggc ccggcaggcg gcttccgtct gggaccggcc ctgggtcaac
     4381 tcattaacca agctcagact gatatcctga gcctcgttaa gccatattta cgttccttgg
     4441 ccgaagaatt agatgaatcc gtttgcttag ccagtttggc tggtgacaag atttatgttt
     4501 tagatcgcat tgtctcggaa cgtgaattgc gtgtcgtttt cccaatcggc atcaatgttc
     4561 cggcggcggc caccgccgcg ggtaaggttt tgctggccgc gctgccagat gagactctcc
     4621 aggcagcctt aggtgaacaa ttgcccgttc tgacatcaaa tacgctgggg cgtaaagctt
     4681 tggtaaagca gctcagtgag gtccgccaat ccggagtggc atcagacctt gatgaacata
     4741 tcgacggtgt ctgtagtttt gcgaccctgc tggacacgta tctgggttac tatagcctgg
     4801 caattgttat gccttccagc cgtgcttcga aacaaagcga tctcattaag aaggcacttt
     4861 tacaatcgaa acttaacatt gaacgcgcga ttggtcgtgc cagcaagaag gccccgtgag
     4921 tcagtttcac ctgttttacg taaaaacccg cttcggcggg tttttacttt tggtactggt
     4981 atggaagcta tacgttacca attgacaaaa aaccacatag tgatacacta ttctgcaatg
     5041 cgggccatgc attgtgtcac acaggaaaca tgacctaagg aggtaaataa tgagaaaggg
     5101 agaggagttg tttactggcg tcgtgcccat tttagtcgag ttagatggag atgtaaatgg
     5161 gcataagttt agtgtacggg gcgagggaga gggcgatgcc acaaatggaa agcttacttt
     5221 aaagtttatt tgtacaacgg ggaagttacc agtaccttgg cctacacttg ttacaacttt
     5281 aacgtatgga gtccaatgtt ttgcacggta tccagatcat atgaagcaac atgatttctt
     5341 taagtcggcc atgccagagg gatatgtcca agagcgcaca ataagcttta aggatgatgg
     5401 aacatataag acacgggcag aggtgaagtt tgagggagat acacttgtaa atcgcataga
     5461 gttgaagggc attgatttta aggaagatgg caacattctg gggcataagt tagagtataa
     5521 ctttaacagc cacaatgtat atattacggc ggacaagcaa aagaatggga taaaggccaa
     5581 ttttaagatt cgccataatg tcgaggatgg gtccgtccaa ctcgccgatc attatcaaca
     5641 aaatactccc attggcgatg gcccagtctt gctccctgat aatcattatc tctcaactca
     5701 atcagtactc tcaaaggatc ctaatgagaa gcgggatcat atggtccttt tagagtttgt
     5761 cactgcagcc ggcattacac atggcatgga tgagttgtat aagtaactcg gtaaagaggg
     5821 gaaatactag atgatgaaat ctaacaatgc gctcatcgtc atcctcggca ccgtcaccct
     5881 ggatgctgta ggcataggct tggttatgcc ggtactgccg ggcctcttgc gggatatcgt
     5941 ccattccgac agcatcgcca gtcactatgg cgtgctgcta gcgctatatg cgttgatgca
     6001 atttctatgc gcacccgttc tcggagcact gtccgaccgc tttggccgcc gcccagtcct
     6061 gctcgcttcg ctacttggag ccactatcga ctacgcgatc atggcgacca cacccgtcct
     6121 gtggatcctc tacgccggac gcatcgtggc cggcatcacc ggcgccacag gtgcggttgc
     6181 tggcgcctat atcgccgaca tcaccgatgg ggaagatcgg gctcgccact tcgggctcat
     6241 gagcgcttgt ttcggcgtgg gtatggtggc aggccccgtg gccgggggac tgttgggcgc
     6301 catctccttg catgcaccat tccttgcggc ggcggtgctc aacggcctca acctactact
     6361 gggctgcttc ctaatgcagg agtcgcataa gggagagcgt cgaccgatgc ccttgagagc
     6421 cttcaaccca gtcagctcct tccggtgggc gcggggcatg actatcgtcg ccgcacttat
     6481 gactgtcttc tttatcatgc aactcgtagg acaggtgccg gcagcgctct gggtcatttt
     6541 cggcgaggac cgctttcgct ggagcgcgac gatgatcggc ctgtcgcttg cggtattcgg
     6601 aatcttgcac gccctcgctc aagccttcgt cactggtccc gccaccaaac gtttcggcga
     6661 gaagcaggcc attatcgccg gcatggcggc cgacgcgctg ggctacgtct tgctggcgtt
     6721 cgcgacgcga ggctggatgg ccttccccat tatgattctt ctcgcttccg gcggcatcgg
     6781 gatgcccgcg ttgcaggcca tgctgtccag gcaggtagat gacgaccatc agggacagct
     6841 tcaaggatcg ctcgcggctc ttaccagcct aacttcgatc attggaccgc tgatcgtcac
     6901 ggcgatttat gccgcctcgg cgagcacatg gaacgggttg gcatggattg taggcgccgc
     6961 cctatacctt gtctgcctcc ccgcgttgcg tcgcggtgca tggagccggg ccacctcgac
     7021 ctgaaccaaa ttccagaaaa gaggcctccc gaaa
//
