LOCUS       sequence-439542-        7597 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To track the natural SoxS 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: SoxS-GFP
FEATURES             Location/Qualifiers
     source          1..7597
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    802..1052
                     /gene="incC"
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     CDS             complement(1055..1810)
                     /codon_start=1
                     /gene="repE"
                     /product="replication initiation protein for the bacterial 
                     F plasmid"
                     /label=repE
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     rep_origin      1901..2120
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     rep_origin      2188..2719
                     /label=oriV
                     /note="incP origin of replication"
     rep_origin      2196..2810
                     /label=oriV
                     /note="origin of replication for the bacterial F plasmid"
     promoter        3668..3770
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             3771..4430
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(3837..3856)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      complement(4565..4659)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(4722..5435)
                     /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(4762..4782)
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     5377..5405
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     RBS             5441..5449
                     /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      5683..5769
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(5927..6898)
                     /codon_start=1
                     /gene="sopB"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopB
                     /translation="MKRAPVIPKHTLNTQPVEDTSLSTPAAPMVDSLIARVGVMARGNA
                     ITLPVCGRDVKFTLEVLRGDSVEKTSRVWSGNERDQELLTEDALDDLIPSFLLTGQQTP
                     AFGRRVSGVIEIADGSRRRKAAALTESDYRVLVGELDDEQMAALSRLGNDYRPTSAYER
                     GQRYASRLQNEFAGNISALADAENISRKIITRCINTAKLPKSVVALFSHPGELSARSGD
                     ALQKAFTDKEELLKQQASNLHEQKKAGVIFEAEEVITLLTSVLKTSSASRTSLSSRHQF
                     APGATVLYKGDKMVLNLDRSRVPTECIEKIEAILKELEKPAP"
     CDS             complement(join(6898..7597,1..476))
                     /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 catccgtgat acattgaggc tgttccctgg gggtcgttac cttccacgag caaaacacgt
       61 agccccttca gagccagatc ctgagcaaga tgaacagaaa ctgaggtttt gtaaacgcca
      121 cctttatggg cagcaacccc gatcaccggt ggaaatacgt cttcagcacg tcgcaatcgc
      181 gtaccaaaca catcacgcat atgattaatt tgttcaattg tataaccaac acgttgctca
      241 acccgtcctc gaatttccat atccgggtgc ggtagtcgcc ctgctttctc ggcatctctg
      301 atagcctgag aagaaacccc aactaaatcc gctgcttcac ctattctcca gcgccgggtt
      361 attttcctcg cttccgggct gtcatcatta aactgtgcaa tggcgatagc cttcgtcatt
      421 tcatgaccag cgtttatgca ctggttaagt gtttccatga gtttcattct gaacatcctt
      481 taatcattgc tttgcgtttt tttattaaat cttgcaattt actgcaaagc aacaacaaaa
      541 tcgcaaagtc atcaaaaaac cgcaaagttg tttaaaataa gagcaacact acaaaaggag
      601 ataagaagag cacatacctc agtcacttat tatcactagc gctcgccgca gccgtgtaac
      661 cgagcatagc gagcgaactg gcgaggaagc aaagaagaac tgttctgtca gatagctctt
      721 acgctcagcg caagaagaaa tatccaccgt gggaaaaact ccaggtagag gtacacacgc
      781 ggatagccaa ttcagagtaa taaactgtga taatcaaccc tcatcaatga tgacgaacta
      841 acccccgata tcaagtcaca tgacgaaggg aaagagaagg aaatcaactg tgacaaactg
      901 ccctcaaatt tggcttcctt aaaaattaca gttcaaaaag tatgagaaaa tccatgcagg
      961 ctgaaggaaa cagcaaaact gtgacaaatt accctcagta ggtcagaaca aatgtgacga
     1021 accaccctca aatctgtgac agataaccct cagactatcc tgtcgtcatg gaagtgatat
     1081 cgcggaagga aaatacgata tgagtcgtct ggcggccttt ctttttctca atgtatgaga
     1141 ggcgcattgg agttctgctg ttgatctcat taacacagac ctgcaggaag cggcggcgga
     1201 agtcaggcat acgctggtaa ctttgaggca gctggtaacg ctctatgatc cagtcgattt
     1261 tcagagagac gatgcctgag ccatccggct tacgatactg acacagggat tcgtataaac
     1321 gcatggcata cggattggtg atttcttttg tttcactaag ccgaaactgc gtaaaccggt
     1381 tctgtaaccc gataaagaag ggaatgagat atgggttgat atgtacactg taaagccctc
     1441 tggatggact gtgcgcacgt ttgataaacc aaggaaaaga ttcatagcct ttttcatcgc
     1501 cggcatcctc ttcagggcga taaaaaacca cttccttccc cgcgaaactc ttcaatgcct
     1561 gccgtatatc cttactggct tccgcagagg tcaatccgaa tatttcagca tatttagcaa
     1621 catggatctc gcagataccg tcatgttcct gtagggtgcc atcagatttt ctgatctggt
     1681 caacgaacag atacagcata cgtttttgat cccgggagag actatatgcc gcctcagtga
     1741 ggtcgtttga ctggacgatt cgcgggctat ttttacgttt cttgtgattg ataaccgctg
     1801 tttccgccat gacagatcca tgtgaagtgt gacaagtttt tagattgtca cactaaataa
     1861 aaaagagtca ataagcaggg ataactttgt gaaaaaacag cttcttctga gggcaatttg
     1921 tcacagggtt aagggcaatt tgtcacagac aggactgtca tttgagggtg atttgtcaca
     1981 ctgaaagggc aatttgtcac aacaccttct ctagaaccag catggataaa ggcctacaag
     2041 gcgctctaaa aaagaagatc taaaaactat aaaaaaaata attataaaaa tatccccgtg
     2101 gataagtgga taaccccaag ggaagttttt tcaggcatcg tgtgtaagca gaatatataa
     2161 gtgctgttcc ctggtgcttc ctcgctcact cgaccgggag ggttcgagaa gggggggcac
     2221 cccccttcgg cgtgcgcggt cacgcgcaca gggcgcagcc ctggttaaaa acaaggttta
     2281 taaatattgg tttaaaagca ggttaaaaga caggttagcg gtggccgaaa aacgggcgga
     2341 aacccttgca aatgctggat tttctgcctg tggacagccc ctcaaatgtc aataggtgcg
     2401 cccctcatct gtcagcactc tgcccctcaa gtgtcaagga tcgcgcccct catctgtcag
     2461 tagtcgcgcc cctcaagtgt caataccgca gggcacttat ccccaggctt gtccacatca
     2521 tctgtgggaa actcgcgtaa aatcaggcgt tttcgccgat ttgcgaggct ggccagctcc
     2581 acgtcgccgg ccgaaatcga gcctgcccct catctgtcaa cgccgcgccg ggtgagtcgg
     2641 cccctcaagt gtcaacgtcc gcccctcatc tgtcagtgag ggccaagttt tccgcgaggt
     2701 atccacaacg ccggcggccg gccgcggtgt ctcgcacacg gcttcgacgg cgtttctggc
     2761 gcgtttgcag ggccatagac ggccgccagc ccagcggcga gggcaaccag ccgagggctt
     2821 cgccctgtcg ctcgactgcg gcgagcacta ctggctgtaa aaggacagac cacatcatgg
     2881 ttctgtgttc attaggttgt tctgtccatt gctgacataa tccgctccac ttcaacgtaa
     2941 caccgcacga agatttctat tgttcctgaa ggcatattca aatcgttttc gttaccgctt
     3001 gctggcatca tgacagaaca ctacttccta taaacgctac acaggctcct gagattaata
     3061 atgcggatct ctacgataat gggagatttt cccgactgtt tcgttcgctt ctcagtggat
     3121 aacagccagc ttctctgttt aacagacaaa aacagcatat ccactcagtt ccacatttcc
     3181 atataaaggc caaggcattt attctcagga taattgtttc agcatcgcaa ccgcatcaga
     3241 ctccggcatc gcaaactgca cccggtgccg ggcagccaca tccagcgcaa aaaccttcgt
     3301 gtagacttcc gttgaactga tggacttatg tcccatcagg ctttgcagaa ctttcagcgg
     3361 tataccggca tacagcatgt gcatcgcata ggaatggcgg aacgtatgtg gtgtgaccgg
     3421 aacagagaac gtcacaccgt cagcagcagc ggcggcaacc gcctccccaa tccaggtcct
     3481 gaccgttctg tccgtcactt cccagatccg cgctttctct gtccttcctg tgcgacggtt
     3541 acgccgctcc atgagcttat cgcgaataaa tacctgtgac ggaagatcac ttcgcagaat
     3601 aaataaatcc tggtgtccct gttgataccg ggaagccctg ggccaacttt tggcgaaaat
     3661 gagacgttga tcggcacgta agaggttcca actttcacca taatgaaata agatcactac
     3721 cgggcgtatt ttttgagtta tcgagatttt caggagctaa ggaagctaaa atggagaaaa
     3781 aaatcactgg atataccacc gttgatatat cccaatggca tcgtaaagaa cattttgagg
     3841 catttcagtc agttgctcaa tgtacctata accagaccgt tcagctggat attacggcct
     3901 ttttaaagac cgtaaagaaa aataagcaca agttttatcc ggcctttatt cacattcttg
     3961 cccgcctgat gaatgctcat ccggaatttc gtatggcaat gaaagacggt gagctggtga
     4021 tatgggatag tgttcaccct tgttacaccg ttttccatga gcaaactgaa acgttttcat
     4081 cgctctggag tgaataccac gacgatttcc ggcagtttct acacatatat tcgcaagatg
     4141 tggcgtgtta cggtgaaaac ctggcctatt tccctaaagg gtttattgag aatatgtttt
     4201 tcgtctcagc caatccctgg gtgagtttca ccagttttga tttaaacgtg gccaatatgg
     4261 acaacttctt cgcccccgtt ttcacgatgg gcaaatatta tacgcaaggc gacaaggtgc
     4321 tgatgccgct ggcgattcag gttcatcatg ccgtttgtga tggcttccat gtcggcagaa
     4381 tgcttaatga attacaacag tactgcgatg agtggcaggg cggggcgtaa tttttttaag
     4441 gcagttattg gtgcccttaa acgcctggtt gctacgcctg aataagtgat aataagcgga
     4501 tgaatggcag aaattcgatg ataagctgtc aaacatgaga attggtctcg acgagtatgc
     4561 ttggattctc accaataaaa aacgcccggc ggcaaccgag cgttctgaac aaatccagat
     4621 ggagttctga ggtcattact ggatctatca acaggagtcc aagcgacgtc tgtgcaagta
     4681 ctactgttct gcagtcactt gaattcgata cccagctgtt atttgtatag ttcatccatg
     4741 ccatgtgtaa tcccagcagc ggttacaaac tcaagaagga ccatgtggtc tctcttttcg
     4801 ttgggatctt tcgaaagggc agattgtgtg gacaggtaat ggttgtctgg taaaaggaca
     4861 gggccatcgc caatcggagt attttgttga taatggtctg ctagttgaac gcttccatct
     4921 tcaatgttgt gtctaatttt gaagttaact ttgattccat tcttttgttt gtctgccatg
     4981 atgtatacat tgtgtgagtt atagttgtat tccaatttgt gtccaagaat gtttccatct
     5041 tctttaaaat caataccttt taactcgatt ctattaacaa gggtatcacc ttcaaacttg
     5101 acttcagcac gtgtcttgta gttcccgtca tctttgaaaa atatagttct ttcctgtacg
     5161 taaccttcgg gcatggcact cttgaaaaag tcatgctgtt tcatgtgatc tgggtatctc
     5221 gcaaagcatt gaacaccata accgaaagta gtgacaagtg ttggccacgg aacaggtagt
     5281 tttccagtag tgcaaataaa tttaagggta agttttccgt atgttgcatc accttcaccc
     5341 tctccactga cagaaaattt gtgcccatta acatcaccat ctaattcaac aagaattggg
     5401 acaactccag tgaaaagttc ttctccttta cgcatattat acctccttaa tttagtgtca
     5461 tctacaactg gcatgatttt attcattgtt taaatacctc cgaggcagaa attacgtcat
     5521 cagacgtcgc taatccatga ctttacgttg ttttacaccc cctgacgcat gtttgcagcc
     5581 tgaatcgtaa actctctatc gttgaatcgc gacagaaaga ttttgggagc aaatgcctag
     5641 ggaccgtctc gagagaatca atattaatcc aacgcgtggc atcaaataaa acgaaaggct
     5701 cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt
     5761 aggacaaatc cgccgcccta gacttaggcg ttcggctgcg gcgagcggta tcagctcact
     5821 caaagaagtg gggcccagag agaatattca ggccagttat gctttctggc ctgtaacaaa
     5881 ggacattaag taaagacaga taaacgtaga ctaaaacgtg gtcgcatcag ggtgctggct
     5941 tttcaagttc cttaagaatg gcctcaattt tctctataca ctcagttgga acacgggacc
     6001 tgtccaggtt aagcaccatt ttatcgccct tatacaatac tgtcgctcca ggagcaaact
     6061 gatgtcgtga gcttaaacta gttcttgatg cagatgacgt tttaagcaca gaagttaaaa
     6121 gagtgataac ttcttcagct tcaaatatca ccccagcttt tttctgctca tgaaggttag
     6181 atgcctgctg cttaagtaat tcctctttat ctgtaaaggc tttttgaagt gcatcacctg
     6241 accgggcaga tagttcaccg gggtgagaaa aaagagcaac aactgattta ggcaatttgg
     6301 cggtgttgat acagcgggta ataatcttac gtgaaatatt ttccgcatca gccagcgcag
     6361 aaatatttcc agcaaattca ttctgcaatc ggcttgcata acgctgacca cgttcataag
     6421 cacttgttgg gcgataatcg ttacccaatc tggataatgc agccatctgc tcatcatcca
     6481 gctcgccaac cagaacacga taatcacttt cggtaagtgc agcagcttta cgacggcgac
     6541 tcccatcggc aatttctatg acaccagata ctcttcgacc gaacgccggt gtctgttgac
     6601 cagtcagtag aaaagaaggg atgagatcat ccagtgcgtc ctcagtaagc agctcctggt
     6661 cacgttcatt acctgaccat acccgagagg tcttctcaac actatcaccc cggagcactt
     6721 caagagtaaa cttcacatcc cgaccacata caggcaaagt aatggcatta ccgcgagcca
     6781 ttactcctac gcgcgcaatt aacgaatcca ccatcggggc agctggtgtc gataacgaag
     6841 tatcttcaac cggttgagta ttgagcgtat gttttggaat aacaggcgca cgcttcatta
     6901 tctaatctcc cagcgtggtt taatcagacg atcgaaaatt tcattgcaga caggttccca
     6961 aatagaaaga gcatttctcc aggcaccagt tgaagagcgt tgatcaatgg cctgttcaaa
     7021 aacagttctc atccggatct gacctttacc aacttcatcc gtttcacgta caacattttt
     7081 tagaaccatg cttccccagg catcccgaat ttgctcctcc atccacgggg actgagagcc
     7141 attactattg ctgtatttgg taagcaaaat acgtacatca ggctcgaacc ctttaagatc
     7201 aacgttcttg agcagatcac gaagcatatc gaaaaactgc agtgcggagg tgtagtcaaa
     7261 caactcagca ggcgtgggaa caatcagcac atcagcagca catacgacat taatcgtgcc
     7321 gatacccagg ttaggcgcgc tgtcaataac tatgacatca tagtcatgag caacagtttc
     7381 aatggccagt cggagcatca ggtgtggatc ggtgggcagt ttaccttcat caaatttgcc
     7441 cattaactca gtttcaatac ggtgcagagc cagacaggaa ggaataatgt caagccccgg
     7501 ccagcaagtg ggctttattg cataagtgac atcgtccttt tccccaagat agaaaggcag
     7561 gagagtgtct tctgcatgaa tatgaagatc tggtacc
//
