LOCUS       sequence-204070-        6624 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  LexA DNA binding domain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6624)
  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 6624)
  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: pHJ2: LexA DBD
FEATURES             Location/Qualifiers
     source          1..6624
                     /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)."
     terminator      1731..1802
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1818..1845
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             2086..3168
                     /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(2105..2124)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     terminator      3284..3315
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             complement(3414..4229)
                     /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     4137..4156
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4221..4240
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             complement(4587..5537)
                     /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      5585..5807
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(6299..6393)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     terminator      complement(6549..6578)
                     /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 taataaactt tatctgagaa tagtcaatct tcggaaatcc caggtggcat
     1681 gctaaaagtc tcgtaaagcg ttctatcaat aacccgttgg tgccaggcat caaataaaac
     1741 gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc
     1801 tctactagag tcacactggc tcaccttcgg gtgggccttt ctgcgtttat accgtctcag
     1861 aatcggccgt gaacaataaa atagtttcgg tttgcattta aatcttacat atgtaatact
     1921 ttcaaagact acatttgtaa gatttgatgt ttgagtcggc tgaaagatcg tacgtaccaa
     1981 ttattgtttc gtgattgttc aagccataac actgtaggga tagtggaaag agtgcttcat
     2041 ctggttacga tcaatcaaat attcaaacgg agggagacga ttttgatgaa accagtaacg
     2101 ttatacgatg tcgcagagta tgccggtgtc tcttatcaga ccgtttcccg cgtggtgaac
     2161 caggccagcc acgtttctgc gaaaacgcgg gaaaaagtgg aagcggcgat ggcggagctg
     2221 aattacattc ccaaccgcgt ggcacaacaa ctggcgggca aacagtcgtt gctgattggc
     2281 gttgccacct ccagtctggc cctgcacgcg ccgtcgcaaa ttgtcgcggc gattaaatct
     2341 cgcgccgatc aactgggtgc cagcgtggtg gtgtcgatgg tagaacgaag cggcgtcgaa
     2401 gcctgtaaaa cggcggtgca caatcttctc gcgcaacgcg tcagtgggct gatcattaac
     2461 tatccgctgg atgaccagga tgccattgct gtggaagctg cctgcactaa tgttccggcg
     2521 ttatttcttg atgtctctga ccagacaccc atcaacagta ttattttctc ccatgaagac
     2581 ggtacgcgac tgggcgtgga gcatctggtc gcattgggtc accagcaaat cgcgctgtta
     2641 gcgggcccat taagttctgt ctcggcgcgt ctgcgtctgg ctggctggca taaatatctc
     2701 actcgcaatc aaattcagcc gatagcggaa cgggaaggcg actggagtgc catgtccggt
     2761 tttcaacaaa ccatgcaaat gctgaatgag ggcatcgttc ccactgcgat gctggttgcc
     2821 aacgatcaga tggcgctggg cgcaatgcgc gccattaccg agtccgggct gcgcgttggt
     2881 gcggatatct cggtagtggg atacgacgat accgaagaca gctcatgtta tatcccgccg
     2941 ttaaccacca tcaaacagga ttttcgcctg ctggggcaaa ccagcgtgga ccgcttgctg
     3001 caactctctc agggccaggc ggtgaagggc aatcagctgt tgcccgtctc actggtgaaa
     3061 agaaaaacca ccctggcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     3121 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgata ataaaaggtc
     3181 ccgtctgaac ttactgtgaa ttcgactaat tattgaccac ttccgagtag aatcgtgctt
     3241 cagtaagaga gtcactaagg gttagttagt tagattagca gaaagtcaaa agcctccgac
     3301 cggaggcttt tgactaaaac ttcccttggg gttatcattg gggctcactc aaaggcggta
     3361 atcagataaa aaaaatcctt agctttcgct aaggatgatt tctgctagta ttattagaaa
     3421 aactcatcga gcatcaaatg aaactgcaat ttattcatat caggattatc aataccatat
     3481 ttttgaaaaa gccgtttctg taatgaagga gaaaactcac cgaggcagtt ccaaagaatg
     3541 gcaaggtcct ggtaacggtc tgcgattccg acccgtccaa catcaataca acctattaat
     3601 ttcccctcgt caaaaataag gttatcaagt gagaaatcac catgagtgac gactgaatcc
     3661 ggtgagaatg gcaagagctt gtgcatttct ttccagactt gttcaacagg ccagccatta
     3721 cgctcgtcat caaaatcact cgcatcaacc aaaccgttat tcatgcgtga ttgcgcctga
     3781 gcaagacgaa atacacgatc gctgttaaaa ggacaattac aaacaggaat cgaatgtaac
     3841 cggcgcagga acacggccag cgcatcaaca atattttcac ctgaatcagg atattcttct
     3901 aatacctgga aggctgtttt cccaggaatc gcggtggtga gtaaccacgc atcatcagga
     3961 gtacggataa aatgcttgat ggtcgggaga ggcataaact ccgtcagcca gttgagacgg
     4021 accatctcat ctgtaacatc attggcaacg ctacctttgc catgtttcag aaacaactct
     4081 ggcgcatcgg gcttcccata caagcgatag attgtcgcac ctgattgccc gacattatcg
     4141 cgagcccatt tatacccata taaatcagcg tccatgttgg agtttaagcg cggacgggag
     4201 caagacgttt cccgttgaat atggctcata acaccccttg tattactgtt tatgtaagca
     4261 gacagtttta ttgttcatga tgatatattt ttatcttgtg caatgtaaca tcagagattt
     4321 tgagacacaa cgtggctttg ttgaataaat cgaacttttg ctgagttgaa ggatcagctc
     4381 tagtagttac attgtcgatc tgttcatggt gaacagcttt aaatgcacca aaaactcgta
     4441 aaagctctga tgtatctatc ttttttacac cgttttcatc tgtgcatatg gacagttttc
     4501 cctttgatat ctaacggtga acagttgttc tacttttgtt tgttagtctt gatgcttcac
     4561 tgatagatac aagagccata agaacctcag atccttccgt atttagccag tatgttctct
     4621 agtgtggttc gttgtttttg cgtgagccat gagaacgaac cattgagatc atgcttactt
     4681 tgcatgtcac tcaaaaattt tgcctcaaaa ctggtgagct gaatttttgc agttaaagca
     4741 tcgtgtagtg tttttcttag tccgttacgt aggtaggaat ctgatgtaat ggttgttggt
     4801 attttgtcac cattcatttt tatctggttg ttctcaagtt cggttacgag atccatttgt
     4861 ctatctagtt caacttggaa aatcaacgta tcagtcgggc ggcctcgctt atcaaccacc
     4921 aatttcatat tgctgtaagt gtttaaatct ttacttattg gtttcaaaac ccattggtta
     4981 agccttttaa actcatggta gttattttca agcattaaca tgaacttaaa ttcatcaagg
     5041 ctaatctcta tatttgcctt gtgagttttc ttttgtgtta gttcttttaa taaccactca
     5101 taaatcctca tagagtattt gttttcaaaa gacttaacat gttccagatt atattttatg
     5161 aattttttta actggaaaag ataaggcaat atctcttcac taaaaactaa ttctaatttt
     5221 tcgcttgaga acttggcata gtttgtccac tggaaaatct caaagccttt aaccaaagga
     5281 ttcctgattt ccacagttct cgtcatcagc tctctggttg ctttagctaa tacaccataa
     5341 gcattttccc tactgatgtt catcatctga gcgtattggt tataagtgaa cgataccgtc
     5401 cgttctttcc ttgtagggtt ttcaatcgtg gggttgagta gtgccacaca gcataaaatt
     5461 agcttggttt catgctccgt taagtcatag cgactaatcg ctagttcatt tgctttgaaa
     5521 acaactaatt cagacataca tctcaattgg tctaggtgat tttaatcact ataccaattg
     5581 agatgggcta gtcaatgata attacatgtc cttttccttt gagttgtggg tatctgtaaa
     5641 ttctgctaga cctttgctgg aaaacttgta aattctgcta gaccctctgt aaattccgct
     5701 agacctttgt gtgttttttt tgtttatatt caagtggtta taatttatag aataaagaaa
     5761 gaataaaaaa agataaaaag aatagatccc agccctgtgt ataactcact actttagtca
     5821 gttccgcagt attacaaaag gatgtcgcaa acgctgtttg ctcctctaca aaacagacct
     5881 taaaacccta aaggcttaag tagcaccctc gcaagctcgg gcaaatcgct gaatattcct
     5941 tttgtctccg accatcaggc acctgagtcg ctgtcttttt cgtgacattc agttcgctgc
     6001 gctcacggct ctggcagtga atgggggtaa atggcactac aggcgccttt tatggattca
     6061 tgcaaggaaa ctacccataa tacaagaaaa gcccgtcacg ggcttctcag ggcgttttat
     6121 ggcgggtctg ctatgtggtg ctatctgact ttttgctgtt cagcagttcc tgccctctga
     6181 ttttccagtc tgaccacttc ggattatccc gtgacaggtc attcagactg gctaatgcac
     6241 ccagtaaggc agcggtatca tcaacaggct tacccgtctt actgtcccta gtgcttggat
     6301 tctcaccaat aaaaaacgcc cggcggcaac cgagcgttct gaacaaatcc agatggagtt
     6361 ctgaggtcat tactggatct atcaacagga gtccaagcga gctcgtaaac ttggtctgac
     6421 agctctagct ccggcaaaaa aacgggcaag gtgtcaccac cctgcccttt ttctttaaaa
     6481 ccgaaaagat tacttcgcgt ttgccacctg acgtctaaga aaaggaatat tcagcaattt
     6541 gcccgtgccg aagaaaggcc cacccgtgaa ggtgagccag tgagttgatt gctacgtaat
     6601 tagttagtta gcccttagtg actc
//
