LOCUS       sequence-436628-        7545 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To track the natural LexA promoter's transcriptional dynamics.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7545)
  AUTHORS   Dash S, Jagadeesan R, Baptista ISC, Chauhan V, Kandavalli V, 
            Oliveira SMD, Ribeiro AS
  TITLE     A library of reporters of the global regulators of gene expression 
            in Escherichia coli.
  JOURNAL   mSystems. 2024 Jun 18;9(6):e0006524. doi: 10.1128/msystems.00065-24.
            Epub 2024 Apr 30.
  PUBMED    38687030
REFERENCE   2  (bases 1 to 7545)
  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: "mSystems.";
            date: "2024-06-18"; volume: "9(6):e0006524. doi"; pages: " 
            10.1128/msystems.00065-24. Epub 2024 Apr 30"
COMMENT     Sequence Label: LexA-GFP
FEATURES             Location/Qualifiers
     source          1..7545
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(659..1834)
                     /codon_start=1
                     /gene="sopA"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopA
                     /translation="MFRMKLMETLNQCINAGHEMTKAIAIAQFNDDSPEARKITRRWRI
                     GEAADLVGVSSQAIRDAEKAGRLPHPDMEIRGRVEQRVGYTIEQINHMRDVFGTRLRRA
                     EDVFPPVIGVAAHKGGVYKTSVSVHLAQDLALKGLRVLLVEGNDPQGTASMYHGWVPDL
                     HIHAEDTLLPFYLGEKDDVTYAIKPTCWPGLDIIPSCLALHRIETELMGKFDEGKLPTD
                     PHLMLRLAIETVAHDYDVIVIDSAPNLGIGTINVVCAADVLIVPTPAELFDYTSALQFF
                     DMLRDLLKNVDLKGFEPDVRILLTKYSNSNGSQSPWMEEQIRDAWGSMVLKNVVRETDE
                     VGKGQIRMRTVFEQAIDQRSSTGAWRNALSIWEPVCNEIFDRLIKPRWEIR"
     misc_feature    2160..2410
                     /gene="incC"
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     CDS             complement(2413..3168)
                     /codon_start=1
                     /gene="repE"
                     /product="replication initiation protein for the bacterial 
                     F plasmid"
                     /label=repE
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     rep_origin      3259..3478
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     rep_origin      3546..4077
                     /label=oriV
                     /note="incP origin of replication"
     rep_origin      3554..4168
                     /label=oriV
                     /note="origin of replication for the bacterial F plasmid"
     promoter        5026..5128
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             5129..5788
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(5195..5214)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      complement(5923..6017)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(6080..6793)
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFVRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(6120..6140)
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     6735..6763
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     RBS             6799..6807
                     /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)"
     terminator      6989..7075
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(join(7233..7545,1..659))
                     /codon_start=1
                     /gene="sopB"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopB
                     /translation="MKRAPVIPKHTLNTQPVEDTSLSTPAAPMVDSLIARVGVMARGNA
                     ITLPVCGRDVKFTLEVLRGDSVEKTSRVWSGNERDQELLTEDALDDLIPSFLLTGQQTP
                     AFGRRVSGVIEIADGSRRRKAAALTESDYRVLVGELDDEQMAALSRLGNDYRPTSAYER
                     GQRYASRLQNEFAGNISALADAENISRKIITRCINTAKLPKSVVALFSHPGELSARSGD
                     ALQKAFTDKEELLKQQASNLHEQKKAGVIFEAEEVITLLTSVLKTSSASRTSLSSRHQF
                     APGATVLYKGDKMVLNLDRSRVPTECIEKIEAILKELEKPAP"
ORIGIN
        1 gaccgggcag atagttcacc ggggtgagaa aaaagagcaa caactgattt aggcaatttg
       61 gcggtgttga tacagcgggt aataatctta cgtgaaatat tttccgcatc agccagcgca
      121 gaaatatttc cagcaaattc attctgcaat cggcttgcat aacgctgacc acgttcataa
      181 gcacttgttg ggcgataatc gttacccaat ctggataatg cagccatctg ctcatcatcc
      241 agctcgccaa ccagaacacg ataatcactt tcggtaagtg cagcagcttt acgacggcga
      301 ctcccatcgg caatttctat gacaccagat actcttcgac cgaacgccgg tgtctgttga
      361 ccagtcagta gaaaagaagg gatgagatca tccagtgcgt cctcagtaag cagctcctgg
      421 tcacgttcat tacctgacca tacccgagag gtcttctcaa cactatcacc ccggagcact
      481 tcaagagtaa acttcacatc ccgaccacat acaggcaaag taatggcatt accgcgagcc
      541 attactccta cgcgcgcaat taacgaatcc accatcgggg cagctggtgt cgataacgaa
      601 gtatcttcaa ccggttgagt attgagcgta tgttttggaa taacaggcgc acgcttcatt
      661 atctaatctc ccagcgtggt ttaatcagac gatcgaaaat ttcattgcag acaggttccc
      721 aaatagaaag agcatttctc caggcaccag ttgaagagcg ttgatcaatg gcctgttcaa
      781 aaacagttct catccggatc tgacctttac caacttcatc cgtttcacgt acaacatttt
      841 ttagaaccat gcttccccag gcatcccgaa tttgctcctc catccacggg gactgagagc
      901 cattactatt gctgtatttg gtaagcaaaa tacgtacatc aggctcgaac cctttaagat
      961 caacgttctt gagcagatca cgaagcatat cgaaaaactg cagtgcggag gtgtagtcaa
     1021 acaactcagc aggcgtggga acaatcagca catcagcagc acatacgaca ttaatcgtgc
     1081 cgatacccag gttaggcgcg ctgtcaataa ctatgacatc atagtcatga gcaacagttt
     1141 caatggccag tcggagcatc aggtgtggat cggtgggcag tttaccttca tcaaatttgc
     1201 ccattaactc agtttcaata cggtgcagag ccagacagga aggaataatg tcaagccccg
     1261 gccagcaagt gggctttatt gcataagtga catcgtcctt ttccccaaga tagaaaggca
     1321 ggagagtgtc ttctgcatga atatgaagat ctggtaccca tccgtgatac attgaggctg
     1381 ttccctgggg gtcgttacct tccacgagca aaacacgtag ccccttcaga gccagatcct
     1441 gagcaagatg aacagaaact gaggttttgt aaacgccacc tttatgggca gcaaccccga
     1501 tcaccggtgg aaatacgtct tcagcacgtc gcaatcgcgt accaaacaca tcacgcatat
     1561 gattaatttg ttcaattgta taaccaacac gttgctcaac ccgtcctcga atttccatat
     1621 ccgggtgcgg tagtcgccct gctttctcgg catctctgat agcctgagaa gaaaccccaa
     1681 ctaaatccgc tgcttcacct attctccagc gccgggttat tttcctcgct tccgggctgt
     1741 catcattaaa ctgtgcaatg gcgatagcct tcgtcatttc atgaccagcg tttatgcact
     1801 ggttaagtgt ttccatgagt ttcattctga acatccttta atcattgctt tgcgtttttt
     1861 tattaaatct tgcaatttac tgcaaagcaa caacaaaatc gcaaagtcat caaaaaaccg
     1921 caaagttgtt taaaataaga gcaacactac aaaaggagat aagaagagca catacctcag
     1981 tcacttatta tcactagcgc tcgccgcagc cgtgtaaccg agcatagcga gcgaactggc
     2041 gaggaagcaa agaagaactg ttctgtcaga tagctcttac gctcagcgca agaagaaata
     2101 tccaccgtgg gaaaaactcc aggtagaggt acacacgcgg atagccaatt cagagtaata
     2161 aactgtgata atcaaccctc atcaatgatg acgaactaac ccccgatatc aagtcacatg
     2221 acgaagggaa agagaaggaa atcaactgtg acaaactgcc ctcaaatttg gcttccttaa
     2281 aaattacagt tcaaaaagta tgagaaaatc catgcaggct gaaggaaaca gcaaaactgt
     2341 gacaaattac cctcagtagg tcagaacaaa tgtgacgaac caccctcaaa tctgtgacag
     2401 ataaccctca gactatcctg tcgtcatgga agtgatatcg cggaaggaaa atacgatatg
     2461 agtcgtctgg cggcctttct ttttctcaat gtatgagagg cgcattggag ttctgctgtt
     2521 gatctcatta acacagacct gcaggaagcg gcggcggaag tcaggcatac gctggtaact
     2581 ttgaggcagc tggtaacgct ctatgatcca gtcgattttc agagagacga tgcctgagcc
     2641 atccggctta cgatactgac acagggattc gtataaacgc atggcatacg gattggtgat
     2701 ttcttttgtt tcactaagcc gaaactgcgt aaaccggttc tgtaacccga taaagaaggg
     2761 aatgagatat gggttgatat gtacactgta aagccctctg gatggactgt gcgcacgttt
     2821 gataaaccaa ggaaaagatt catagccttt ttcatcgccg gcatcctctt cagggcgata
     2881 aaaaaccact tccttccccg cgaaactctt caatgcctgc cgtatatcct tactggcttc
     2941 cgcagaggtc aatccgaata tttcagcata tttagcaaca tggatctcgc agataccgtc
     3001 atgttcctgt agggtgccat cagattttct gatctggtca acgaacagat acagcatacg
     3061 tttttgatcc cgggagagac tatatgccgc ctcagtgagg tcgtttgact ggacgattcg
     3121 cgggctattt ttacgtttct tgtgattgat aaccgctgtt tccgccatga cagatccatg
     3181 tgaagtgtga caagttttta gattgtcaca ctaaataaaa aagagtcaat aagcagggat
     3241 aactttgtga aaaaacagct tcttctgagg gcaatttgtc acagggttaa gggcaatttg
     3301 tcacagacag gactgtcatt tgagggtgat ttgtcacact gaaagggcaa tttgtcacaa
     3361 caccttctct agaaccagca tggataaagg cctacaaggc gctctaaaaa agaagatcta
     3421 aaaactataa aaaaaataat tataaaaata tccccgtgga taagtggata accccaaggg
     3481 aagttttttc aggcatcgtg tgtaagcaga atatataagt gctgttccct ggtgcttcct
     3541 cgctcactcg accgggaggg ttcgagaagg gggggcaccc cccttcggcg tgcgcggtca
     3601 cgcgcacagg gcgcagccct ggttaaaaac aaggtttata aatattggtt taaaagcagg
     3661 ttaaaagaca ggttagcggt ggccgaaaaa cgggcggaaa cccttgcaaa tgctggattt
     3721 tctgcctgtg gacagcccct caaatgtcaa taggtgcgcc cctcatctgt cagcactctg
     3781 cccctcaagt gtcaaggatc gcgcccctca tctgtcagta gtcgcgcccc tcaagtgtca
     3841 ataccgcagg gcacttatcc ccaggcttgt ccacatcatc tgtgggaaac tcgcgtaaaa
     3901 tcaggcgttt tcgccgattt gcgaggctgg ccagctccac gtcgccggcc gaaatcgagc
     3961 ctgcccctca tctgtcaacg ccgcgccggg tgagtcggcc cctcaagtgt caacgtccgc
     4021 ccctcatctg tcagtgaggg ccaagttttc cgcgaggtat ccacaacgcc ggcggccggc
     4081 cgcggtgtct cgcacacggc ttcgacggcg tttctggcgc gtttgcaggg ccatagacgg
     4141 ccgccagccc agcggcgagg gcaaccagcc gagggcttcg ccctgtcgct cgactgcggc
     4201 gagcactact ggctgtaaaa ggacagacca catcatggtt ctgtgttcat taggttgttc
     4261 tgtccattgc tgacataatc cgctccactt caacgtaaca ccgcacgaag atttctattg
     4321 ttcctgaagg catattcaaa tcgttttcgt taccgcttgc tggcatcatg acagaacact
     4381 acttcctata aacgctacac aggctcctga gattaataat gcggatctct acgataatgg
     4441 gagattttcc cgactgtttc gttcgcttct cagtggataa cagccagctt ctctgtttaa
     4501 cagacaaaaa cagcatatcc actcagttcc acatttccat ataaaggcca aggcatttat
     4561 tctcaggata attgtttcag catcgcaacc gcatcagact ccggcatcgc aaactgcacc
     4621 cggtgccggg cagccacatc cagcgcaaaa accttcgtgt agacttccgt tgaactgatg
     4681 gacttatgtc ccatcaggct ttgcagaact ttcagcggta taccggcata cagcatgtgc
     4741 atcgcatagg aatggcggaa cgtatgtggt gtgaccggaa cagagaacgt cacaccgtca
     4801 gcagcagcgg cggcaaccgc ctccccaatc caggtcctga ccgttctgtc cgtcacttcc
     4861 cagatccgcg ctttctctgt ccttcctgtg cgacggttac gccgctccat gagcttatcg
     4921 cgaataaata cctgtgacgg aagatcactt cgcagaataa ataaatcctg gtgtccctgt
     4981 tgataccggg aagccctggg ccaacttttg gcgaaaatga gacgttgatc ggcacgtaag
     5041 aggttccaac tttcaccata atgaaataag atcactaccg ggcgtatttt ttgagttatc
     5101 gagattttca ggagctaagg aagctaaaat ggagaaaaaa atcactggat ataccaccgt
     5161 tgatatatcc caatggcatc gtaaagaaca ttttgaggca tttcagtcag ttgctcaatg
     5221 tacctataac cagaccgttc agctggatat tacggccttt ttaaagaccg taaagaaaaa
     5281 taagcacaag ttttatccgg cctttattca cattcttgcc cgcctgatga atgctcatcc
     5341 ggaatttcgt atggcaatga aagacggtga gctggtgata tgggatagtg ttcacccttg
     5401 ttacaccgtt ttccatgagc aaactgaaac gttttcatcg ctctggagtg aataccacga
     5461 cgatttccgg cagtttctac acatatattc gcaagatgtg gcgtgttacg gtgaaaacct
     5521 ggcctatttc cctaaagggt ttattgagaa tatgtttttc gtctcagcca atccctgggt
     5581 gagtttcacc agttttgatt taaacgtggc caatatggac aacttcttcg cccccgtttt
     5641 cacgatgggc aaatattata cgcaaggcga caaggtgctg atgccgctgg cgattcaggt
     5701 tcatcatgcc gtttgtgatg gcttccatgt cggcagaatg cttaatgaat tacaacagta
     5761 ctgcgatgag tggcagggcg gggcgtaatt tttttaaggc agttattggt gcccttaaac
     5821 gcctggttgc tacgcctgaa taagtgataa taagcggatg aatggcagaa attcgatgat
     5881 aagctgtcaa acatgagaat tggtctcgac gagtatgctt ggattctcac caataaaaaa
     5941 cgcccggcgg caaccgagcg ttctgaacaa atccagatgg agttctgagg tcattactgg
     6001 atctatcaac aggagtccaa gcgacgtctg tgcaagtact actgttctgc agtcacttga
     6061 attcgatacc cagctgttat ttgtatagtt catccatgcc atgtgtaatc ccagcagcgg
     6121 ttacaaactc aagaaggacc atgtggtctc tcttttcgtt gggatctttc gaaagggcag
     6181 attgtgtgga caggtaatgg ttgtctggta aaaggacagg gccatcgcca atcggagtat
     6241 tttgttgata atggtctgct agttgaacgc ttccatcttc aatgttgtgt ctaattttga
     6301 agttaacttt gattccattc ttttgtttgt ctgccatgat gtatacattg tgtgagttat
     6361 agttgtattc caatttgtgt ccaagaatgt ttccatcttc tttaaaatca atacctttta
     6421 actcgattct attaacaagg gtatcacctt caaacttgac ttcagcacgt gtcttgtagt
     6481 tcccgtcatc tttgaaaaat atagttcttt cctgtacgta accttcgggc atggcactct
     6541 tgaaaaagtc atgctgtttc atgtgatctg ggtatctcac aaagcattga acaccataac
     6601 cgaaagtagt gacaagtgtt ggccacggaa caggtagttt tccagtagtg caaataaatt
     6661 taagggtaag ttttccgtat gttgcatcac cttcaccctc tccactgaca gaaaatttgt
     6721 gcccattaac atcaccatct aattcaacaa gaattgggac aactccagtg aaaagttctt
     6781 ctcctttacg catattatac ctccttaatt tagtgtcatc tacaactggc attccgcccc
     6841 ctgggtgtat atacagttat gctgtgagta tatacagcaa aaggcgattt tggaaccata
     6901 aactgcacaa taaaccagag atttatcgaa ttctggaagg gcctagggac cgtctcgaga
     6961 gaatcaatat taatccaacg cgtggcatca aataaaacga aaggctcagt cgaaagactg
     7021 ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc ctgagtagga caaatccgcc
     7081 gccctagact taggcgttcg gctgcggcga gcggtatcag ctcactcaaa gaagtggggc
     7141 ccagagagaa tattcaggcc agttatgctt tctggcctgt aacaaaggac attaagtaaa
     7201 gacagataaa cgtagactaa aacgtggtcg catcagggtg ctggcttttc aagttcctta
     7261 agaatggcct caattttctc tatacactca gttggaacac gggacctgtc caggttaagc
     7321 accattttat cgcccttata caatactgtc gctccaggag caaactgatg tcgtgagctt
     7381 aaactagttc ttgatgcaga tgacgtttta agcacagaag ttaaaagagt gataacttct
     7441 tcagcttcaa atatcacccc agcttttttc tgctcatgaa ggttagatgc ctgctgctta
     7501 agtaattcct ctttatctgt aaaggctttt tgaagtgcat cacct
//
