LOCUS       sequence-203310-        6999 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  sfGFP expression repressed by transcriptional factor LexA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6999)
  AUTHORS   Chang HJ, Mayonove P, Zavala A, De Visch A, Minard P, Cohen-Gonsaud 
            M, Bonnet J
  TITLE     A Modular Receptor Platform To Expand the Sensing Repertoire of 
            Bacteria.
  JOURNAL   ACS Synth Biol. 2017 Oct 30. doi: 10.1021/acssynbio.7b00266.
  PUBMED    28946740
REFERENCE   2  (bases 1 to 6999)
  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"; journalName: "ACS Synth 
            Biol. 2017 Oct 30. doi: 10.1021/acssynbio.7b00266."
COMMENT     Sequence Label: pHJ1_Full length LexA
FEATURES             Location/Qualifiers
     source          1..6999
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(294..1007,1008..1010))
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /note="mammalian codon-optimized"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      1157..1184
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     protein_bind    1287..1303
                     /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    1310..1326
                     /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)."
     CDS             1976..2005
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     terminator      2106..2177
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2193..2220
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             2461..3543
                     /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
                     EACKTAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(2480..2499)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     terminator      3659..3690
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             complement(3789..4604)
                     /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"
     primer_bind     4512..4531
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4596..4615
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             complement(4962..5912)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLRNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKHDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      5960..6182
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(6674..6768)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     terminator      complement(6924..6953)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
ORIGIN
        1 ctcggatacc cttactctgt tgaaaacgaa tagataggtt tgctcgtagt ttaccacgga
       61 tacagacagt gataatctta ggcgcgccga atcttgcttg gcaaggttct tttttttttt
      121 caaaaaagca cgaaaaaagc acccataagg taaacttttg agtgctttta aatacgcttc
      181 acagtttctt cttcattttt aacaaaaaac ccagctcatt gagctgggtt taagctggct
      241 tggcggcgtc ctaagattac tactgataac cactgttgat tgggataccc gtattattta
      301 taaagttcgt ccataccgtg agtgatacct gctgctgtaa caaattcaag aagtaccata
      361 tggtcacgtt tttcgttagg atcttttgaa agaactgatt gagtagaaag gtagtggtta
      421 tctggaagca atactggtcc gtctccgatt ggtgtatttt gttgataatg atcagccaat
      481 tgaacagatc catcttcaac attgtgacga attttaaaat tagctttaat accgtttttt
      541 tgtttgtcag ctgtgatata aacattatga gagttgaagt tgtattcaag tttgtgtccc
      601 aaaatgttac catcttcttt gaaatcgatt cctttcaatt cgatacgatt aacaagagtg
      661 tcaccttcga attttacttc agcacgtgtt ttatatgttc cgtcatcttt aaatgaaatt
      721 gtacgttctt gtacataacc ttctggcata gcagatttaa aaaaatcgtg acgtttcata
      781 tgatcaggat aacgtgaaaa acattgtact ccatatgtca atgttgtaac aagagtaggc
      841 caaggaacag gaagttttcc agttgtacaa atgaatttaa gtgtcaattt accgttagtt
      901 gcatcacctt caccttcacc acgtacagaa aatttgtgac cgttaacatc accatccaat
      961 tcaaccaaga taggtactac acctgtaaaa agttcttctc cttttgacat ttcaccacct
     1021 ttactagtcc agcccctgtg agttaccgta tggatgtacg gtacatccag tgacaacaaa
     1081 gatcaaccct attttcggaa agagcctcga aggaacggtt atttctgcgt agatctatct
     1141 tacacagcat cacactggct caccttcggg tgggcctttc tgcgtttata tactagagag
     1201 agaatataaa aagccagatt attaatccgg cttttttatt atttaggcaa ctgaaacgat
     1261 tcggatcctg tattactatt cttaaattgt gagcggataa caattgacat tgtgagcgga
     1321 taacaagata ctgagcacag ctagcgatta actaataagg aggacaaaca tgaaagcgct
     1381 gaccgcgcgt caacaagaag ttttcgacct gatccgtgac cacatcagcc agacgggtat
     1441 gccacctacc cgtgcggaaa tcgcgcaacg tctgggcttc cgtagcgcaa gctctgctga
     1501 agagcacctg aaagcgctgg cccgtaaggg tgttatcgaa atcgtttccg gtgcgagccg
     1561 cggtatccgt ctgctgcagg aagaagaaga aggtctgccg ctggtgggcc gtgtggctgc
     1621 cggcgaacca ctgctggccc agcaacacat tgagggccat tatcaagtag atccgagcct
     1681 gttcaagcct aatgccgact ttctgctgcg tgtgagcggc atgagcatga aagacattgg
     1741 catcatggat ggcgatctgc tggcagtcca taagacgcag gacgtacgca atggccaagt
     1801 ggtggttgca cgcattgacg atgaggttac cgtaaaacgc ctgaaaaagc agggcaacaa
     1861 ggtcgaactg ctgccggaaa atagcgagtt taagccgatt gttgtcgatc tgcgccaaca
     1921 gtcctttacg atcgaaggtc tggctgtcgg cgtcattcgc aacggcgatt ggctggaaca
     1981 aaaactgatt tctgaggaag atctgtaata aactttatct gagaatagtc aatcttcgga
     2041 aatcccaggt ggcatgctaa aagtctcgta aagcgttcta tcaataaccc gttggtgcca
     2101 ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt
     2161 tgtcggtgaa cgctctctac tagagtcaca ctggctcacc ttcgggtggg cctttctgcg
     2221 tttataccgt ctcagaatcg gccgtgaaca ataaaatagt ttcggtttgc atttaaatct
     2281 tacatatgta atactttcaa agactacatt tgtaagattt gatgtttgag tcggctgaaa
     2341 gatcgtacgt accaattatt gtttcgtgat tgttcaagcc ataacactgt agggatagtg
     2401 gaaagagtgc ttcatctggt tacgatcaat caaatattca aacggaggga gacgattttg
     2461 atgaaaccag taacgttata cgatgtcgca gagtatgccg gtgtctctta tcagaccgtt
     2521 tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa cgcgggaaaa agtggaagcg
     2581 gcgatggcgg agctgaatta cattcccaac cgcgtggcac aacaactggc gggcaaacag
     2641 tcgttgctga ttggcgttgc cacctccagt ctggccctgc acgcgccgtc gcaaattgtc
     2701 gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg tggtggtgtc gatggtagaa
     2761 cgaagcggcg tcgaagcctg taaaacggcg gtgcacaatc ttctcgcgca acgcgtcagt
     2821 gggctgatca ttaactatcc gctggatgac caggatgcca ttgctgtgga agctgcctgc
     2881 actaatgttc cggcgttatt tcttgatgtc tctgaccaga cacccatcaa cagtattatt
     2941 ttctcccatg aagacggtac gcgactgggc gtggagcatc tggtcgcatt gggtcaccag
     3001 caaatcgcgc tgttagcggg cccattaagt tctgtctcgg cgcgtctgcg tctggctggc
     3061 tggcataaat atctcactcg caatcaaatt cagccgatag cggaacggga aggcgactgg
     3121 agtgccatgt ccggttttca acaaaccatg caaatgctga atgagggcat cgttcccact
     3181 gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa tgcgcgccat taccgagtcc
     3241 gggctgcgcg ttggtgcgga tatctcggta gtgggatacg acgataccga agacagctca
     3301 tgttatatcc cgccgttaac caccatcaaa caggattttc gcctgctggg gcaaaccagc
     3361 gtggaccgct tgctgcaact ctctcagggc caggcggtga agggcaatca gctgttgccc
     3421 gtctcactgg tgaaaagaaa aaccaccctg gcgcccaata cgcaaaccgc ctctccccgc
     3481 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
     3541 tgataataaa aggtcccgtc tgaacttact gtgaattcga ctaattattg accacttccg
     3601 agtagaatcg tgcttcagta agagagtcac taagggttag ttagttagat tagcagaaag
     3661 tcaaaagcct ccgaccggag gcttttgact aaaacttccc ttggggttat cattggggct
     3721 cactcaaagg cggtaatcag ataaaaaaaa tccttagctt tcgctaagga tgatttctgc
     3781 tagtattatt agaaaaactc atcgagcatc aaatgaaact gcaatttatt catatcagga
     3841 ttatcaatac catatttttg aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg
     3901 cagttccaaa gaatggcaag gtcctggtaa cggtctgcga ttccgacccg tccaacatca
     3961 atacaaccta ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa atcaccatga
     4021 gtgacgactg aatccggtga gaatggcaag agcttgtgca tttctttcca gacttgttca
     4081 acaggccagc cattacgctc gtcatcaaaa tcactcgcat caaccaaacc gttattcatg
     4141 cgtgattgcg cctgagcaag acgaaataca cgatcgctgt taaaaggaca attacaaaca
     4201 ggaatcgaat gtaaccggcg caggaacacg gccagcgcat caacaatatt ttcacctgaa
     4261 tcaggatatt cttctaatac ctggaaggct gttttcccag gaatcgcggt ggtgagtaac
     4321 cacgcatcat caggagtacg gataaaatgc ttgatggtcg ggagaggcat aaactccgtc
     4381 agccagttga gacggaccat ctcatctgta acatcattgg caacgctacc tttgccatgt
     4441 ttcagaaaca actctggcgc atcgggcttc ccatacaagc gatagattgt cgcacctgat
     4501 tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcgtccat gttggagttt
     4561 aagcgcggac gggagcaaga cgtttcccgt tgaatatggc tcataacacc ccttgtatta
     4621 ctgtttatgt aagcagacag ttttattgtt catgatgata tatttttatc ttgtgcaatg
     4681 taacatcaga gattttgaga cacaacgtgg ctttgttgaa taaatcgaac ttttgctgag
     4741 ttgaaggatc agctctagta gttacattgt cgatctgttc atggtgaaca gctttaaatg
     4801 caccaaaaac tcgtaaaagc tctgatgtat ctatcttttt tacaccgttt tcatctgtgc
     4861 atatggacag ttttcccttt gatatctaac ggtgaacagt tgttctactt ttgtttgtta
     4921 gtcttgatgc ttcactgata gatacaagag ccataagaac ctcagatcct tccgtattta
     4981 gccagtatgt tctctagtgt ggttcgttgt ttttgcgtga gccatgagaa cgaaccattg
     5041 agatcatgct tactttgcat gtcactcaaa aattttgcct caaaactggt gagctgaatt
     5101 tttgcagtta aagcatcgtg tagtgttttt cttagtccgt tacgtaggta ggaatctgat
     5161 gtaatggttg ttggtatttt gtcaccattc atttttatct ggttgttctc aagttcggtt
     5221 acgagatcca tttgtctatc tagttcaact tggaaaatca acgtatcagt cgggcggcct
     5281 cgcttatcaa ccaccaattt catattgctg taagtgttta aatctttact tattggtttc
     5341 aaaacccatt ggttaagcct tttaaactca tggtagttat tttcaagcat taacatgaac
     5401 ttaaattcat caaggctaat ctctatattt gccttgtgag ttttcttttg tgttagttct
     5461 tttaataacc actcataaat cctcatagag tatttgtttt caaaagactt aacatgttcc
     5521 agattatatt ttatgaattt ttttaactgg aaaagataag gcaatatctc ttcactaaaa
     5581 actaattcta atttttcgct tgagaacttg gcatagtttg tccactggaa aatctcaaag
     5641 cctttaacca aaggattcct gatttccaca gttctcgtca tcagctctct ggttgcttta
     5701 gctaatacac cataagcatt ttccctactg atgttcatca tctgagcgta ttggttataa
     5761 gtgaacgata ccgtccgttc tttccttgta gggttttcaa tcgtggggtt gagtagtgcc
     5821 acacagcata aaattagctt ggtttcatgc tccgttaagt catagcgact aatcgctagt
     5881 tcatttgctt tgaaaacaac taattcagac atacatctca attggtctag gtgattttaa
     5941 tcactatacc aattgagatg ggctagtcaa tgataattac atgtcctttt cctttgagtt
     6001 gtgggtatct gtaaattctg ctagaccttt gctggaaaac ttgtaaattc tgctagaccc
     6061 tctgtaaatt ccgctagacc tttgtgtgtt ttttttgttt atattcaagt ggttataatt
     6121 tatagaataa agaaagaata aaaaaagata aaaagaatag atcccagccc tgtgtataac
     6181 tcactacttt agtcagttcc gcagtattac aaaaggatgt cgcaaacgct gtttgctcct
     6241 ctacaaaaca gaccttaaaa ccctaaaggc ttaagtagca ccctcgcaag ctcgggcaaa
     6301 tcgctgaata ttccttttgt ctccgaccat caggcacctg agtcgctgtc tttttcgtga
     6361 cattcagttc gctgcgctca cggctctggc agtgaatggg ggtaaatggc actacaggcg
     6421 ccttttatgg attcatgcaa ggaaactacc cataatacaa gaaaagcccg tcacgggctt
     6481 ctcagggcgt tttatggcgg gtctgctatg tggtgctatc tgactttttg ctgttcagca
     6541 gttcctgccc tctgattttc cagtctgacc acttcggatt atcccgtgac aggtcattca
     6601 gactggctaa tgcacccagt aaggcagcgg tatcatcaac aggcttaccc gtcttactgt
     6661 ccctagtgct tggattctca ccaataaaaa acgcccggcg gcaaccgagc gttctgaaca
     6721 aatccagatg gagttctgag gtcattactg gatctatcaa caggagtcca agcgagctcg
     6781 taaacttggt ctgacagctc tagctccggc aaaaaaacgg gcaaggtgtc accaccctgc
     6841 cctttttctt taaaaccgaa aagattactt cgcgtttgcc acctgacgtc taagaaaagg
     6901 aatattcagc aatttgcccg tgccgaagaa aggcccaccc gtgaaggtga gccagtgagt
     6961 tgattgctac gtaattagtt agttagccct tagtgactc
//
