LOCUS       sequence-441279-        7597 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To track the natural CpxR promoter's transcriptional dynamics.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7597)
  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 7597)
  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: CpxR-GFP
FEATURES             Location/Qualifiers
     source          1..7597
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             210..1181
                     /codon_start=1
                     /gene="sopB"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopB
                     /translation="MKRAPVIPKHTLNTQPVEDTSLSTPAAPMVDSLIARVGVMARGNA
                     ITLPVCGRDVKFTLEVLRGDSVEKTSRVWSGNERDQELLTEDALDDLIPSFLLTGQQTP
                     AFGRRVSGVIEIADGSRRRKAAALTESDYRVLVGELDDEQMAALSRLGNDYRPTSAYER
                     GQRYASRLQNEFAGNISALADAENISRKIITRCINTAKLPKSVVALFSHPGELSARSGD
                     ALQKAFTDKEELLKQQASNLHEQKKAGVIFEAEEVITLLTSVLKTSSASRTSLSSRHQF
                     APGATVLYKGDKMVLNLDRSRVPTECIEKIEAILKELEKPAP"
     terminator      complement(1339..1425)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     RBS             1659..1667
                     /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             1673..2386
                     /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
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(1703..1731)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2326..2346
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      2449..2543
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2678..3337)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3252..3271
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(3338..3440)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     rep_origin      4298..4912
                     /label=oriV
                     /note="origin of replication for the bacterial F plasmid"
     rep_origin      4389..4920
                     /label=oriV
                     /note="incP origin of replication"
     rep_origin      4988..5207
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     CDS             5298..6053
                     /codon_start=1
                     /gene="repE"
                     /product="replication initiation protein for the bacterial 
                     F plasmid"
                     /label=repE
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     misc_feature    6056..6306
                     /gene="incC"
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     CDS             join(6632..7597,1..210)
                     /codon_start=1
                     /gene="sopA"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopA
                     /translation="MFRMKLMETLNQCINAGHEMTKAIAIAQFNDDSPEARKITRRWRI
                     GEAADLVGVSSQAIRDAEKAGRLPHPDMEIRGRVEQRVGYTIEQINHMRDVFGTRLRRA
                     EDVFPPVIGVAAHKGGVYKTSVSVHLAQDLALKGLRVLLVEGNDPQGTASMYHGWVPDL
                     HIHAEDTLLPFYLGEKDDVTYAIKPTCWPGLDIIPSCLALHRIETELMGKFDEGKLPTD
                     PHLMLRLAIETVAHDYDVIVIDSAPNLGIGTINVVCAADVLIVPTPAELFDYTSALQFF
                     DMLRDLLKNVDLKGFEPDVRILLTKYSNSNGSQSPWMEEQIRDAWGSMVLKNVVRETDE
                     VGKGQIRMRTVFEQAIDQRSSTGAWRNALSIWEPVCNEIFDRLIKPRWEIR"
ORIGIN
        1 cgggatgcct ggggaagcat ggttctaaaa aatgttgtac gtgaaacgga tgaagttggt
       61 aaaggtcaga tccggatgag aactgttttt gaacaggcca ttgatcaacg ctcttcaact
      121 ggtgcctgga gaaatgctct ttctatttgg gaacctgtct gcaatgaaat tttcgatcgt
      181 ctgattaaac cacgctggga gattagataa tgaagcgtgc gcctgttatt ccaaaacata
      241 cgctcaatac tcaaccggtt gaagatactt cgttatcgac accagctgcc ccgatggtgg
      301 attcgttaat tgcgcgcgta ggagtaatgg ctcgcggtaa tgccattact ttgcctgtat
      361 gtggtcggga tgtgaagttt actcttgaag tgctccgggg tgatagtgtt gagaagacct
      421 ctcgggtatg gtcaggtaat gaacgtgacc aggagctgct tactgaggac gcactggatg
      481 atctcatccc ttcttttcta ctgactggtc aacagacacc ggcgttcggt cgaagagtat
      541 ctggtgtcat agaaattgcc gatgggagtc gccgtcgtaa agctgctgca cttaccgaaa
      601 gtgattatcg tgttctggtt ggcgagctgg atgatgagca gatggctgca ttatccagat
      661 tgggtaacga ttatcgccca acaagtgctt atgaacgtgg tcagcgttat gcaagccgat
      721 tgcagaatga atttgctgga aatatttctg cgctggctga tgcggaaaat atttcacgta
      781 agattattac ccgctgtatc aacaccgcca aattgcctaa atcagttgtt gctctttttt
      841 ctcaccccgg tgaactatct gcccggtcag gtgatgcact tcaaaaagcc tttacagata
      901 aagaggaatt acttaagcag caggcatcta accttcatga gcagaaaaaa gctggggtga
      961 tatttgaagc tgaagaagtt atcactcttt taacttctgt gcttaaaacg tcatctgcat
     1021 caagaactag tttaagctca cgacatcagt ttgctcctgg agcgacagta ttgtataagg
     1081 gcgataaaat ggtgcttaac ctggacaggt cccgtgttcc aactgagtgt atagagaaaa
     1141 ttgaggccat tcttaaggaa cttgaaaagc cagcaccctg atgcgaccac gttttagtct
     1201 acgtttatct gtctttactt aatgtccttt gttacaggcc agaaagcata actggcctga
     1261 atattctctc tgggccccac ttctttgagt gagctgatac cgctcgccgc agccgaacgc
     1321 ctaagtctag ggcggcggat ttgtcctact caggagagcg ttcaccgaca aacaacagat
     1381 aaaacgaaag gcccagtctt tcgactgagc ctttcgtttt atttgatgcc acgcgttgga
     1441 ttaatattga ttctctcgag acggtcccta ggcatttgct cccaaaatct ttctgtcgcg
     1501 attcaacgat agagagttta cgattcaggc tgcaaacatg cgtcaggggg tgtaaaacaa
     1561 cgtaaagtca tggattagcg acgtctgatg acgtaatttc tgcctcggag gtatttaaac
     1621 aatgaataaa atcatgccag ttgtagatga cactaaatta aggaggtata atatgcgtaa
     1681 aggagaagaa cttttcactg gagttgtccc aattcttgtt gaattagatg gtgatgttaa
     1741 tgggcacaaa ttttctgtca gtggagaggg tgaaggtgat gcaacatacg gaaaacttac
     1801 ccttaaattt atttgcacta ctggaaaact acctgttccg tggccaacac ttgtcactac
     1861 tttcggttat ggtgttcaat gctttgcgag atacccagat cacatgaaac agcatgactt
     1921 tttcaagagt gccatgcccg aaggttacgt acaggaaaga actatatttt tcaaagatga
     1981 cgggaactac aagacacgtg ctgaagtcaa gtttgaaggt gatacccttg ttaatagaat
     2041 cgagttaaaa ggtattgatt ttaaagaaga tggaaacatt cttggacaca aattggaata
     2101 caactataac tcacacaatg tatacatcat ggcagacaaa caaaagaatg gaatcaaagt
     2161 taacttcaaa attagacaca acattgaaga tggaagcgtt caactagcag accattatca
     2221 acaaaatact ccgattggcg atggccctgt ccttttacca gacaaccatt acctgtccac
     2281 acaatctgcc ctttcgaaag atcccaacga aaagagagac cacatggtcc ttcttgagtt
     2341 tgtaaccgct gctgggatta cacatggcat ggatgaacta tacaaataac agctgggtat
     2401 cgaattcaag tgactgcaga acagtagtac ttgcacagac gtcgcttgga ctcctgttga
     2461 tagatccagt aatgacctca gaactccatc tggatttgtt cagaacgctc ggttgccgcc
     2521 gggcgttttt tattggtgag aatccaagca tactcgtcga gaccaattct catgtttgac
     2581 agcttatcat cgaatttctg ccattcatcc gcttattatc acttattcag gcgtagcaac
     2641 caggcgttta agggcaccaa taactgcctt aaaaaaatta cgccccgccc tgccactcat
     2701 cgcagtactg ttgtaattca ttaagcattc tgccgacatg gaagccatca caaacggcat
     2761 gatgaacctg aatcgccagc ggcatcagca ccttgtcgcc ttgcgtataa tatttgccca
     2821 tcgtgaaaac gggggcgaag aagttgtcca tattggccac gtttaaatca aaactggtga
     2881 aactcaccca gggattggct gagacgaaaa acatattctc aataaaccct ttagggaaat
     2941 aggccaggtt ttcaccgtaa cacgccacat cttgcgaata tatgtgtaga aactgccgga
     3001 aatcgtcgtg gtattcactc cagagcgatg aaaacgtttc agtttgctca tggaaaacgg
     3061 tgtaacaagg gtgaacacta tcccatatca ccagctcacc gtctttcatt gccatacgaa
     3121 attccggatg agcattcatc aggcgggcaa gaatgtgaat aaaggccgga taaaacttgt
     3181 gcttattttt ctttacggtc tttaaaaagg ccgtaatatc cagctgaacg gtctggttat
     3241 aggtacattg agcaactgac tgaaatgcct caaaatgttc tttacgatgc cattgggata
     3301 tatcaacggt ggtatatcca gtgatttttt tctccatttt agcttcctta gctcctgaaa
     3361 atctcgataa ctcaaaaaat acgcccggta gtgatcttat ttcattatgg tgaaagttgg
     3421 aacctcttac gtgccgatca acgtctcatt ttcgccaaaa gttggcccag ggcttcccgg
     3481 tatcaacagg gacaccagga tttatttatt ctgcgaagtg atcttccgtc acaggtattt
     3541 attcgcgata agctcatgga gcggcgtaac cgtcgcacag gaaggacaga gaaagcgcgg
     3601 atctgggaag tgacggacag aacggtcagg acctggattg gggaggcggt tgccgccgct
     3661 gctgctgacg gtgtgacgtt ctctgttccg gtcacaccac atacgttccg ccattcctat
     3721 gcgatgcaca tgctgtatgc cggtataccg ctgaaagttc tgcaaagcct gatgggacat
     3781 aagtccatca gttcaacgga agtctacacg aaggtttttg cgctggatgt ggctgcccgg
     3841 caccgggtgc agtttgcgat gccggagtct gatgcggttg cgatgctgaa acaattatcc
     3901 tgagaataaa tgccttggcc tttatatgga aatgtggaac tgagtggata tgctgttttt
     3961 gtctgttaaa cagagaagct ggctgttatc cactgagaag cgaacgaaac agtcgggaaa
     4021 atctcccatt atcgtagaga tccgcattat taatctcagg agcctgtgta gcgtttatag
     4081 gaagtagtgt tctgtcatga tgccagcaag cggtaacgaa aacgatttga atatgccttc
     4141 aggaacaata gaaatcttcg tgcggtgtta cgttgaagtg gagcggatta tgtcagcaat
     4201 ggacagaaca acctaatgaa cacagaacca tgatgtggtc tgtcctttta cagccagtag
     4261 tgctcgccgc agtcgagcga cagggcgaag ccctcggctg gttgccctcg ccgctgggct
     4321 ggcggccgtc tatggccctg caaacgcgcc agaaacgccg tcgaagccgt gtgcgagaca
     4381 ccgcggccgg ccgccggcgt tgtggatacc tcgcggaaaa cttggccctc actgacagat
     4441 gaggggcgga cgttgacact tgaggggccg actcacccgg cgcggcgttg acagatgagg
     4501 ggcaggctcg atttcggccg gcgacgtgga gctggccagc ctcgcaaatc ggcgaaaacg
     4561 cctgatttta cgcgagtttc ccacagatga tgtggacaag cctggggata agtgccctgc
     4621 ggtattgaca cttgaggggc gcgactactg acagatgagg ggcgcgatcc ttgacacttg
     4681 aggggcagag tgctgacaga tgaggggcgc acctattgac atttgagggg ctgtccacag
     4741 gcagaaaatc cagcatttgc aagggtttcc gcccgttttt cggccaccgc taacctgtct
     4801 tttaacctgc ttttaaacca atatttataa accttgtttt taaccagggc tgcgccctgt
     4861 gcgcgtgacc gcgcacgccg aaggggggtg cccccccttc tcgaaccctc ccggtcgagt
     4921 gagcgaggaa gcaccaggga acagcactta tatattctgc ttacacacga tgcctgaaaa
     4981 aacttccctt ggggttatcc acttatccac ggggatattt ttataattat tttttttata
     5041 gtttttagat cttctttttt agagcgcctt gtaggccttt atccatgctg gttctagaga
     5101 aggtgttgtg acaaattgcc ctttcagtgt gacaaatcac cctcaaatga cagtcctgtc
     5161 tgtgacaaat tgcccttaac cctgtgacaa attgccctca gaagaagctg ttttttcaca
     5221 aagttatccc tgcttattga ctctttttta tttagtgtga caatctaaaa acttgtcaca
     5281 cttcacatgg atctgtcatg gcggaaacag cggttatcaa tcacaagaaa cgtaaaaata
     5341 gcccgcgaat cgtccagtca aacgacctca ctgaggcggc atatagtctc tcccgggatc
     5401 aaaaacgtat gctgtatctg ttcgttgacc agatcagaaa atctgatggc accctacagg
     5461 aacatgacgg tatctgcgag atccatgttg ctaaatatgc tgaaatattc ggattgacct
     5521 ctgcggaagc cagtaaggat atacggcagg cattgaagag tttcgcgggg aaggaagtgg
     5581 ttttttatcg ccctgaagag gatgccggcg atgaaaaagg ctatgaatct tttccttggt
     5641 ttatcaaacg tgcgcacagt ccatccagag ggctttacag tgtacatatc aacccatatc
     5701 tcattccctt ctttatcggg ttacagaacc ggtttacgca gtttcggctt agtgaaacaa
     5761 aagaaatcac caatccgtat gccatgcgtt tatacgaatc cctgtgtcag tatcgtaagc
     5821 cggatggctc aggcatcgtc tctctgaaaa tcgactggat catagagcgt taccagctgc
     5881 ctcaaagtta ccagcgtatg cctgacttcc gccgccgctt cctgcaggtc tgtgttaatg
     5941 agatcaacag cagaactcca atgcgcctct catacattga gaaaaagaaa ggccgccaga
     6001 cgactcatat cgtattttcc ttccgcgata tcacttccat gacgacagga tagtctgagg
     6061 gttatctgtc acagatttga gggtggttcg tcacatttgt tctgacctac tgagggtaat
     6121 ttgtcacagt tttgctgttt ccttcagcct gcatggattt tctcatactt tttgaactgt
     6181 aatttttaag gaagccaaat ttgagggcag tttgtcacag ttgatttcct tctctttccc
     6241 ttcgtcatgt gacttgatat cgggggttag ttcgtcatca ttgatgaggg ttgattatca
     6301 cagtttatta ctctgaattg gctatccgcg tgtgtacctc tacctggagt ttttcccacg
     6361 gtggatattt cttcttgcgc tgagcgtaag agctatctga cagaacagtt cttctttgct
     6421 tcctcgccag ttcgctcgct atgctcggtt acacggctgc ggcgagcgct agtgataata
     6481 agtgactgag gtatgtgctc ttcttatctc cttttgtagt gttgctctta ttttaaacaa
     6541 ctttgcggtt ttttgatgac tttgcgattt tgttgttgct ttgcagtaaa ttgcaagatt
     6601 taataaaaaa acgcaaagca atgattaaag gatgttcaga atgaaactca tggaaacact
     6661 taaccagtgc ataaacgctg gtcatgaaat gacgaaggct atcgccattg cacagtttaa
     6721 tgatgacagc ccggaagcga ggaaaataac ccggcgctgg agaataggtg aagcagcgga
     6781 tttagttggg gtttcttctc aggctatcag agatgccgag aaagcagggc gactaccgca
     6841 cccggatatg gaaattcgag gacgggttga gcaacgtgtt ggttatacaa ttgaacaaat
     6901 taatcatatg cgtgatgtgt ttggtacgcg attgcgacgt gctgaagacg tatttccacc
     6961 ggtgatcggg gttgctgccc ataaaggtgg cgtttacaaa acctcagttt ctgttcatct
     7021 tgctcaggat ctggctctga aggggctacg tgttttgctc gtggaaggta acgaccccca
     7081 gggaacagcc tcaatgtatc acggatgggt accagatctt catattcatg cagaagacac
     7141 tctcctgcct ttctatcttg gggaaaagga cgatgtcact tatgcaataa agcccacttg
     7201 ctggccgggg cttgacatta ttccttcctg tctggctctg caccgtattg aaactgagtt
     7261 aatgggcaaa tttgatgaag gtaaactgcc caccgatcca cacctgatgc tccgactggc
     7321 cattgaaact gttgctcatg actatgatgt catagttatt gacagcgcgc ctaacctggg
     7381 tatcggcacg attaatgtcg tatgtgctgc tgatgtgctg attgttccca cgcctgctga
     7441 gttgtttgac tacacctccg cactgcagtt tttcgatatg cttcgtgatc tgctcaagaa
     7501 cgttgatctt aaagggttcg agcctgatgt acgtattttg cttaccaaat acagcaatag
     7561 taatggctct cagtccccgt ggatggagga gcaaatt
//
