LOCUS       sequence-210108-        6802 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 6802)
  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 6802)
  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..6802
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             650..679
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     terminator      780..851
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      867..894
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             1135..2217
                     /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(1154..1173)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             complement(2369..3184)
                     /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     3092..3111
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3176..3195
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             complement(3542..4492)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      4540..4762
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(5254..5348)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(5770..6483,6484..6486))
                     /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      6633..6660
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     protein_bind    6763..6779
                     /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    6786..6802
                     /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)."
ORIGIN
        1 gatactgagc acagctagcg attaactaat aaggaggaca aacatgaaag cgctgaccgc
       61 gcgtcaacaa gaagttttcg acctgatccg tgaccacatc agccagacgg gtatgccacc
      121 tacccgtgcg gaaatcgcgc aacgtctggg cttccgtagc gcaagctctg ctgaagagca
      181 cctgaaagcg ctggcccgta agggtgttat cgaaatcgtt tccggtgcga gccgcggtat
      241 ccgtctgctg caggaagaag aagaaggtct gccgctggtg ggccgtgtgg ctgccggcga
      301 accactgctg gcccagcaac acattgaggg ccattatcaa gtagatccga gcctgttcaa
      361 gcctaatgcc gactttctgc tgcgtgtgag cggcatgagc atgaaagaca ttggcatcat
      421 ggatggcgat ctgctggcag tccataagac gcaggacgta cgcaatggcc aagtggtggt
      481 tgcacgcatt gacgatgagg ttaccgtaaa acgcctgaaa aagcagggca acaaggtcga
      541 actgctgccg gaaaatagcg agtttaagcc gattgttgtc gatctgcgcc aacagtcctt
      601 tacgatcgaa ggtctggctg tcggcgtcat tcgcaacggc gattggctgg aacaaaaact
      661 gatttctgag gaagatctgt aataaacttt atctgagaat agtcaatctt cggaaatccc
      721 aggtggcatg ctaaaagtct cgtaaagcgt tctatcaata acccgttggt gccaggcatc
      781 aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg
      841 tgaacgctct ctactagagt cacactggct caccttcggg tgggcctttc tgcgtttata
      901 ccgtctcaga atcggccgtg aacaataaaa tagtttcggt ttgcatttaa atcttacata
      961 tgtaatactt tcaaagacta catttgtaag atttgatgtt tgagtcggct gaaagatcgt
     1021 acgtaccaat tattgtttcg tgattgttca agccataaca ctgtagggat agtggaaaga
     1081 gtgcttcatc tggttacgat caatcaaata ttcaaacgga gggagacgat tttgatgaaa
     1141 ccagtaacgt tatacgatgt cgcagagtat gccggtgtct cttatcagac cgtttcccgc
     1201 gtggtgaacc aggccagcca cgtttctgcg aaaacgcggg aaaaagtgga agcggcgatg
     1261 gcggagctga attacattcc caaccgcgtg gcacaacaac tggcgggcaa acagtcgttg
     1321 ctgattggcg ttgccacctc cagtctggcc ctgcacgcgc cgtcgcaaat tgtcgcggcg
     1381 attaaatctc gcgccgatca actgggtgcc agcgtggtgg tgtcgatggt agaacgaagc
     1441 ggcgtcgaag cctgtaaaac ggcggtgcac aatcttctcg cgcaacgcgt cagtgggctg
     1501 atcattaact atccgctgga tgaccaggat gccattgctg tggaagctgc ctgcactaat
     1561 gttccggcgt tatttcttga tgtctctgac cagacaccca tcaacagtat tattttctcc
     1621 catgaagacg gtacgcgact gggcgtggag catctggtcg cattgggtca ccagcaaatc
     1681 gcgctgttag cgggcccatt aagttctgtc tcggcgcgtc tgcgtctggc tggctggcat
     1741 aaatatctca ctcgcaatca aattcagccg atagcggaac gggaaggcga ctggagtgcc
     1801 atgtccggtt ttcaacaaac catgcaaatg ctgaatgagg gcatcgttcc cactgcgatg
     1861 ctggttgcca acgatcagat ggcgctgggc gcaatgcgcg ccattaccga gtccgggctg
     1921 cgcgttggtg cggatatctc ggtagtggga tacgacgata ccgaagacag ctcatgttat
     1981 atcccgccgt taaccaccat caaacaggat tttcgcctgc tggggcaaac cagcgtggac
     2041 cgcttgctgc aactctctca gggccaggcg gtgaagggca atcagctgtt gcccgtctca
     2101 ctggtgaaaa gaaaaaccac cctggcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
     2161 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgataa
     2221 taaaaggtcc cgtctgaact tactgtgaat tcgactaatt attgaccact tccgagtaga
     2281 atcgtgcttc agtaagagct cactcaaagg cggtaatcag ataaaaaaaa tccttagctt
     2341 tcgctaagga tgatttctgc tagtattatt agaaaaactc atcgagcatc aaatgaaact
     2401 gcaatttatt catatcagga ttatcaatac catatttttg aaaaagccgt ttctgtaatg
     2461 aaggagaaaa ctcaccgagg cagttccaaa gaatggcaag gtcctggtaa cggtctgcga
     2521 ttccgacccg tccaacatca atacaaccta ttaatttccc ctcgtcaaaa ataaggttat
     2581 caagtgagaa atcaccatga gtgacgactg aatccggtga gaatggcaag agcttgtgca
     2641 tttctttcca gacttgttca acaggccagc cattacgctc gtcatcaaaa tcactcgcat
     2701 caaccaaacc gttattcatg cgtgattgcg cctgagcaag acgaaataca cgatcgctgt
     2761 taaaaggaca attacaaaca ggaatcgaat gtaaccggcg caggaacacg gccagcgcat
     2821 caacaatatt ttcacctgaa tcaggatatt cttctaatac ctggaaggct gttttcccag
     2881 gaatcgcggt ggtgagtaac cacgcatcat caggagtacg gataaaatgc ttgatggtcg
     2941 ggagaggcat aaactccgtc agccagttga gacggaccat ctcatctgta acatcattgg
     3001 caacgctacc tttgccatgt ttcagaaaca actctggcgc atcgggcttc ccatacaagc
     3061 gatagattgt cgcacctgat tgcccgacat tatcgcgagc ccatttatac ccatataaat
     3121 cagcgtccat gttggagttt aagcgcggac gggagcaaga cgtttcccgt tgaatatggc
     3181 tcataacacc ccttgtatta ctgtttatgt aagcagacag ttttattgtt catgatgata
     3241 tatttttatc ttgtgcaatg taacatcaga gattttgaga cacaacgtgg ctttgttgaa
     3301 taaatcgaac ttttgctgag ttgaaggatc agctctagta gttacattgt cgatctgttc
     3361 atggtgaaca gctttgaatg caccaaaaac tcgtaaaagc tctgatgtat ctatcttttt
     3421 tacaccgttt tcatctgtgc atatggacag ttttcccttt gatatgtaac ggtgaacagt
     3481 tgttctactt ttgtttgtta gtcttgatgc ttcactgata gatacaagag ccataagaac
     3541 ctcagatcct tccgtattta gccagtatgt tctctagtgt ggttcgttgt ttttgcgtga
     3601 gccatgagaa cgaaccattg agatcatact tactttgcat gtcactcaaa aattttgcct
     3661 caaaactggt gagctgaatt tttgcagtta aagcatcgtg tagtgttttt cttagtccgt
     3721 tatgtaggta ggaatctgat gtaatggttg ttggtatttt gtcaccattc atttttatct
     3781 ggttgttctc aagttcggtt acgagatcca tttgtctatc tagttcaact tggaaaatca
     3841 acgtatcagt cgggcggcct cgcttatcaa ccaccaattt catattgctg taagtgttta
     3901 aatctttact tattggtttc aaaacccatt ggttaagcct tttaaactca tggtagttat
     3961 tttcaagcat taacatgaac ttaaattcat caaggctaat ctctatattt gccttgtgag
     4021 ttttcttttg tgttagttct tttaataacc actcataaat cctcatagag tatttgtttt
     4081 caaaagactt aacatgttcc agattatatt ttatgaattt ttttaactgg aaaagataag
     4141 gcaatatctc ttcactaaaa actaattcta atttttcgct tgagaacttg gcatagtttg
     4201 tccactggaa aatctcaaag cctttaacca aaggattcct gatttccaca gttctcgtca
     4261 tcagctctct ggttgcttta gctaatacac cataagcatt ttccctactg atgttcatca
     4321 tctgagcgta ttggttataa gtgaacgata ccgtccgttc tttccttgta gggttttcaa
     4381 tcgtggggtt gagtagtgcc acacagcata aaattagctt ggtttcatgc tccgttaagt
     4441 catagcgact aatcgctagt tcatttgctt tgaaaacaac taattcagac atacatctca
     4501 attggtctag gtgattttaa tcactatacc aattgagatg ggctagtcaa tgataattac
     4561 atgtcctttt cctttgagtt gtgggtatct gtaaattctg ctagaccttt gctggaaaac
     4621 ttgtaaattc tgctagaccc tctgtaaatt ccgctagacc tttgtgtgtt ttttttgttt
     4681 atattcaagt ggttataatt tatagaataa agaaagaata aaaaaagata aaaagaatag
     4741 atcccagccc tgtgtataac tcactacttt agtcagttcc gcagtattac aaaaggatgt
     4801 cgcaaacgct gtttgctcct ctacaaaaca gaccttaaaa ccctaaaggc ttaagtagca
     4861 ccctcgcaag ctcgggcaaa tcgctgaata tttcttttgt ctccgaccat caggcacctg
     4921 agtcgctgtc tttttcgtga cattcagttc gctgcgctca cggctctggc agtgaatggg
     4981 ggtaaatggc actacaggcg ccttttatgg attcatgcaa ggaaactacc cataatacaa
     5041 gaaaagcccg tcacgggctt ctcagggcgt tttatggcgg gtctgctatg tggtgctatc
     5101 tgactttttg ctgttcagca gttcctgccc tctgattttc cagtctgacc acttcggatt
     5161 atcccgtgac aggtcattca gactggctaa tgcacccagt aaggcagcgg tatcatcaac
     5221 aggcttaccc gtcttactgt ccctagtgct tggattctca ccaataaaaa acgcccggcg
     5281 gcaaccgagc gttctgaaca aatccagatg gagttctgag gtcattactg gatctatcaa
     5341 caggagtcca agcgagctcg taaacttggt ctgacagctc tagctccggc aaaaaaacgg
     5401 gcaaggtgtc accaccctgc cctttttctt taaaaccgaa aagattactt cgcgtttgcc
     5461 acctgacgtc taagaactcg gataccctta ctctgttgaa aacgaataga taggtttgct
     5521 cgtagtttac cacggataca gacagtgata atcttaggcg cgccgaatct tgcttggcaa
     5581 ggttcttttt ttttttcaaa aaagcacgaa aaaagcaccc ataaggtaaa cttttgagtg
     5641 cttttaaata cgcttcacag tttcttcttc atttttaaca aaaaacccag ctcattgagc
     5701 tgggtttaag ctggcttggc ggcgtcctaa gattactact gataaccact gttgattggg
     5761 atacccgtat tatttataaa gttcgtccat accgtgagtg atacctgctg ctgtaacaaa
     5821 ttcaagaagt accatatggt cacgtttttc gttaggatct tttgaaagaa ctgattgagt
     5881 agaaaggtag tggttatctg gaagcaatac tggtccgtct ccgattggtg tattttgttg
     5941 ataatgatca gccaattgaa cagatccatc ttcaacattg tgacgaattt taaaattagc
     6001 tttaataccg tttttttgtt tgtcagctgt gatataaaca ttatgagagt tgaagttgta
     6061 ttcaagtttg tgtcccaaaa tgttaccatc ttctttgaaa tcgattcctt tcaattcgat
     6121 acgattaaca agagtgtcac cttcgaattt tacttcagca cgtgttttat atgttccgtc
     6181 atctttaaat gaaattgtac gttcttgtac ataaccttct ggcatagcag atttaaaaaa
     6241 atcgtgacgt ttcatatgat caggataacg tgaaaaacat tgtactccat atgtcaatgt
     6301 tgtaacaaga gtaggccaag gaacaggaag ttttccagtt gtacaaatga atttaagtgt
     6361 caatttaccg ttagttgcat caccttcacc ttcaccacgt acagaaaatt tgtgaccgtt
     6421 aacatcacca tccaattcaa ccaagatagg tactacacct gtaaaaagtt cttctccttt
     6481 tgacatttca ccacctttac tagtccagcc cctgtgagtt accgtatgga tgtacggtac
     6541 atccagtgac aacaaagatc aaccctattt tcggaaagag cctcgaagga acggttattt
     6601 ctgcgtggat ctatcttaca cagcatcaca ctggctcacc ttcgggtggg cctttctgcg
     6661 tttatatact agagagagaa tataaaaagc cagattatta atccggcttt tttattattt
     6721 aggcaactga aacgattcgg atcctgtatt actattctta aattgtgagc ggataacaat
     6781 tgacattgtg agcggataac aa
//
