LOCUS       sequence-436633-        7764 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To track the natural Fis promoter's transcriptional dynamics.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7764)
  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 7764)
  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: Fis-GFP
FEATURES             Location/Qualifiers
     source          1..7764
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      586..805
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     rep_origin      873..1404
                     /label=oriV
                     /note="incP origin of replication"
     rep_origin      881..1495
                     /label=oriV
                     /note="origin of replication for the bacterial F plasmid"
     promoter        2353..2455
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             2456..3115
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2522..2541)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      complement(3250..3344)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3407..4120)
                     /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(3447..3467)
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     4062..4090
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     RBS             4126..4134
                     /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      4535..4621
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(4779..5750)
                     /codon_start=1
                     /gene="sopB"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopB
                     /translation="MKRAPVIPKHTLNTQPVEDTSLSTPAAPMVDSLIARVGVMARGNA
                     ITLPVCGRDVKFTLEVLRGDSVEKTSRVWSGNERDQELLTEDALDDLIPSFLLTGQQTP
                     AFGRRVSGVIEIADGSRRRKAAALTESDYRVLVGELDDEQMAALSRLGNDYRPTSAYER
                     GQRYASRLQNEFAGNISALADAENISRKIITRCINTAKLPKSVVALFSHPGELSARSGD
                     ALQKAFTDKEELLKQQASNLHEQKKAGVIFEAEEVITLLTSVLKTSSASRTSLSSRHQF
                     APGATVLYKGDKMVLNLDRSRVPTECIEKIEAILKELEKPAP"
     CDS             complement(5750..6925)
                     /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    7251..7501
                     /gene="incC"
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     CDS             complement(join(7504..7764,1..495))
                     /codon_start=1
                     /gene="repE"
                     /product="replication initiation protein for the bacterial 
                     F plasmid"
                     /label=repE
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
ORIGIN
        1 taaacgcatg gcatacggat tggtgatttc ttttgtttca ctaagccgaa actgcgtaaa
       61 ccggttctgt aacccgataa agaagggaat gagatatggg ttgatatgta cactgtaaag
      121 ccctctggat ggactgtgcg cacgtttgat aaaccaagga aaagattcat agcctttttc
      181 atcgccggca tcctcttcag ggcgataaaa aaccacttcc ttccccgcga aactcttcaa
      241 tgcctgccgt atatccttac tggcttccgc agaggtcaat ccgaatattt cagcatattt
      301 agcaacatgg atctcgcaga taccgtcatg ttcctgtagg gtgccatcag attttctgat
      361 ctggtcaacg aacagataca gcatacgttt ttgatcccgg gagagactat atgccgcctc
      421 agtgaggtcg tttgactgga cgattcgcgg gctattttta cgtttcttgt gattgataac
      481 cgctgtttcc gccatgacag atccatgtga agtgtgacaa gtttttagat tgtcacacta
      541 aataaaaaag agtcaataag cagggataac tttgtgaaaa aacagcttct tctgagggca
      601 atttgtcaca gggttaaggg caatttgtca cagacaggac tgtcatttga gggtgatttg
      661 tcacactgaa agggcaattt gtcacaacac cttctctaga accagcatgg ataaaggcct
      721 acaaggcgct ctaaaaaaga agatctaaaa actataaaaa aaataattat aaaaatatcc
      781 ccgtggataa gtggataacc ccaagggaag ttttttcagg catcgtgtgt aagcagaata
      841 tataagtgct gttccctggt gcttcctcgc tcactcgacc gggagggttc gagaaggggg
      901 ggcacccccc ttcggcgtgc gcggtcacgc gcacagggcg cagccctggt taaaaacaag
      961 gtttataaat attggtttaa aagcaggtta aaagacaggt tagcggtggc cgaaaaacgg
     1021 gcggaaaccc ttgcaaatgc tggattttct gcctgtggac agcccctcaa atgtcaatag
     1081 gtgcgcccct catctgtcag cactctgccc ctcaagtgtc aaggatcgcg cccctcatct
     1141 gtcagtagtc gcgcccctca agtgtcaata ccgcagggca cttatcccca ggcttgtcca
     1201 catcatctgt gggaaactcg cgtaaaatca ggcgttttcg ccgatttgcg aggctggcca
     1261 gctccacgtc gccggccgaa atcgagcctg cccctcatct gtcaacgccg cgccgggtga
     1321 gtcggcccct caagtgtcaa cgtccgcccc tcatctgtcg gtgagggcca agttttccgc
     1381 gaggtatcca caacgccggc ggccggccgc ggtgtctcgc acacggcttc gacggcgttt
     1441 ctggcgcgtt tgcagggcca tagacggccg ccagcccagc ggcgagggca accagccgag
     1501 ggcttcgccc tgtcgctcga ctgcggcgag cactactggc tgtaaaagga cagaccacat
     1561 catggttctg tgttcattag gttgttctgt ccattgctga cataatccgc tccacttcaa
     1621 cgtaacaccg cacgaagatt tctattgttc ctgaaggcat attcaaatcg ttttcgttac
     1681 cgcttgctgg catcatgaca gaacactact tcctataaac gctacacagg ctcctgagat
     1741 taataatgcg gatctctacg ataatgggag attttcccga ctgtttcgtt cgcttctcag
     1801 tggataacag ccagcttctc tgtttaacag acaaaaacag catatccact cagttccaca
     1861 tttccatata aaggccaagg catttattct caggataatt gtttcagcat cgcaaccgca
     1921 tcagactccg gcatcgcaaa ctgcacccgg tgccgggcag ccacatccag cgcaaaaacc
     1981 ttcgtgtaga cttccgttga actgatggac ttatgtccca tcaggctttg cagaactttc
     2041 agcggtatac cggcatacag catgtgcatc gcataggaat ggcggaacgt atgtggtgtg
     2101 accggaacag agaacgtcac accgtcagca gcagcggcgg caaccgcctc cccaatccag
     2161 gtcctgaccg ttctgtccgt cacttcccag atccgcgctt tctctgtcct tcctgtgcga
     2221 cggttacgcc gctccatgag cttatcgcga ataaatacct gtgacggaag atcacttcgc
     2281 agaataaata aatcctggtg tccctgttga taccgggaag ccctgggcca acttttggcg
     2341 aaaatgagac gttgatcggc acgtaagagg ttccaacttt caccataatg aaataagatc
     2401 actaccgggc gtattttttg agttatcgag attttcagga gctaaggaag ctaaaatgga
     2461 gaaaaaaatc actggatata ccaccgttga tatatcccaa tggcatcgta aagaacattt
     2521 tgaggcattt cagtcagttg ctcaatgtac ctataaccag accgttcagc tggatattac
     2581 ggccttttta aagaccgtaa agaaaaataa gcacaagttt tatccggcct ttattcacat
     2641 tcttgcccgc ctgatgaatg ctcatccgga atttcgtatg gcaatgaaag acggtgagct
     2701 ggtgatatgg gatagtgttc acccttgtta caccgttttc catgagcaaa ctgaaacgtt
     2761 ttcatcgctc tggagtgaat accacgacga tttccggcag tttctacaca tatattcgca
     2821 agatgtggcg tgttacggtg aaaacctggc ctatttccct aaagggttta ttgagaatat
     2881 gtttttcgtc tcagccaatc cctgggtgag tttcaccagt tttgatttaa acgtggccaa
     2941 tatggacaac ttcttcgccc ccgttttcac gatgggcaaa tattatacgc aaggcgacaa
     3001 ggtgctgatg ccgctggcga ttcaggttca tcatgccgtt tgtgatggct tccatgtcgg
     3061 cagaatgctt aatgaattac aacagtactg cgatgagtgg cagggcgggg cgtaattttt
     3121 ttaaggcagt tattggtgcc cttaaacgcc tggttgctac gcctgaataa gtgataataa
     3181 gcggatgaat ggcagaaatt cgatgataag ctgtcaaaca tgagaattgg tctcgacgag
     3241 tatgcttgga ttctcaccaa taaaaaacgc ccggcggcaa ccgagcgttc tgaacaaatc
     3301 cagatggagt tctgaggtca ttactggatc tatcaacagg agtccaagcg acgtctgtgc
     3361 aagtactact gttctgcagt cacttgaatt cgatacccag ctgttatttg tatagttcat
     3421 ccatgccatg tgtaatccca gcagcggtta caaactcaag aaggaccatg tggtctctct
     3481 tttcgttggg atctttcgaa agggcagatt gtgtggacag gtaatggttg tctggtaaaa
     3541 ggacagggcc atcgccaatc ggagtatttt gttgataatg gtctgctagt tgaacgcttc
     3601 catcttcaat gttgtgtcta attttgaagt taactttgat tccattcttt tgtttgtctg
     3661 ccatgatgta tacattgtgt gagttatagt tgtattccaa tttgtgtcca agaatgtttc
     3721 catcttcttt aaaatcaata ccttttaact cgattctatt aacaagggta tcaccttcaa
     3781 acttgacttc agcacgtgtc ttgtagttcc cgtcatcttt gaaaaatata gttctttcct
     3841 gtacgtaacc ttcgggcatg gcactcttga aaaagtcatg ctgtttcatg tgatctgggt
     3901 atctcgcaaa gcattgaaca ccataaccga aagtagtgac aagtgttggc cacggaacag
     3961 gtagttttcc agtagtgcaa ataaatttaa gggtaagttt tccgtatgtt gcatcacctt
     4021 caccctctcc actgacagaa aatttgtgcc cattaacatc accatctaat tcaacaagaa
     4081 ttgggacaac tccagtgaaa agttcttctc ctttacgcat attatacctc cttaatttag
     4141 tgtcatctac aactggcatg agttaagaaa tgaccatact gtgactgcaa ggcggcgtat
     4201 attacgcatt ttttgcacga gatgaaaggc caaactttga ccaatcctct gagatggatc
     4261 aaagaattgc atttaaaatg agcgtggtgc gataattact cataaaaatc atcatattag
     4321 aaaatagtga ctaaaattta cactcaaaga aatttgagta agttctcaat ttttctttat
     4381 gaatgaaaat tttggcacgc aaatttgcgt aaataatcgg caaatttacg tgccttttgt
     4441 gagcttgctc gcacttcgcc ccgcgtcacc ctacggcgat gcgaaggcct agggaccgtc
     4501 tcgagagaat caatattaat ccaacgcgtg gcatcaaata aaacgaaagg ctcagtcgaa
     4561 agactgggcc tttcgtttta tctgttgttt gtcggtgaac gctctcctga gtaggacaaa
     4621 tccgccgccc tagacttagg cgttcggctg cggcgagcgg tatcagctca ctcaaagaag
     4681 tggggcccag agagaatatt caggccagtt atgctttctg gcctgtaaca aaggacatta
     4741 agtaaagaca gataaacgta gactaaaacg tggtcgcatc agggtgctgg cttttcaagt
     4801 tccttaagaa tggcctcaat tttctctata cactcagttg gaacacggga cctgtccagg
     4861 ttaagcacca ttttatcgcc cttatacaat actgtcgctc caggagcaaa ctgatgtcgt
     4921 gagcttaaac tagttcttga tgcagatgac gttttaagca cagaagttaa aagagtgata
     4981 acttcttcag cttcaaatat caccccagct tttttctgct catgaaggtt agatgcctgc
     5041 tgcttaagta attcctcttt atctgtaaag gctttttgaa gtgcatcacc tgaccgggca
     5101 gatagttcac cggggtgaga aaaaagagca acaactgatt taggcaattt ggcggtgttg
     5161 atacagcggg taataatctt acgtgaaata ttttccgcat cagccagcgc agaaatattt
     5221 ccagcaaatt cattctgcaa tcggcttgca taacgctgac cacgttcata agcacttgtt
     5281 gggcgataat cgttacccaa tctggataat gcagccatct gctcatcatc cagctcgcca
     5341 accagaacac gataatcact ttcggtaagt gcagcagctt tacgacggcg actcccatcg
     5401 gcaatttcta tgacaccaga tactcttcga ccgaacgccg gtgtctgttg accagtcagt
     5461 agaaaagaag ggatgagatc atccagtgcg tcctcagtaa gcagctcctg gtcacgttca
     5521 ttacctgacc atacccgaga ggtcttctca acactatcac cccggagcac ttcaagagta
     5581 aacttcacat cccgaccaca tacaggcaaa gtaatggcat taccgcgagc cattactcct
     5641 acgcgcgcaa ttaacgaatc caccatcggg gcagctggtg tcgataacga agtatcttca
     5701 accggttgag tattgagcgt atgttttgga ataacaggcg cacgcttcat tatctaatct
     5761 cccagcgtgg tttaatcaga cgatcgaaaa tttcattgca gacaggttcc caaatagaaa
     5821 gagcatttct ccaggcacca gttgaagagc gttgatcaat ggcctgttca aaaacagttc
     5881 tcatccggat ctgaccttta ccaacttcat ccgtttcacg tacaacattt tttagaacca
     5941 tgcttcccca ggcatcccga atttgctcct ccatccacgg ggactgagag ccattactat
     6001 tgctgtattt ggtaagcaaa atacgtacat caggctcgaa ccctttaaga tcaacgttct
     6061 tgagcagatc acgaagcata tcgaaaaact gcagtgcgga ggtgtagtca aacaactcag
     6121 caggcgtggg aacaatcagc acatcagcag cacatacgac attaatcgtg ccgataccca
     6181 ggttaggcgc gctgtcaata actatgacat catagtcatg agcaacagtt tcaatggcca
     6241 gtcggagcat caggtgtgga tcggtgggca gtttaccttc atcaaatttg cccattaact
     6301 cagtttcaat acggtgcaga gccagacagg aaggaataat gtcaagcccc ggccagcaag
     6361 tgggctttat tgcataagtg acatcgtcct tttccccaag atagaaaggc aggagagtgt
     6421 cttctgcatg aatatgaaga tctggtaccc atccgtgata cattgaggct gttccctggg
     6481 ggtcgttacc ttccacgagc aaaacacgta gccccttcag agccagatcc tgagcaagat
     6541 gaacagaaac tgaggttttg taaacgccac ctttatgggc agcaaccccg atcaccggtg
     6601 gaaatacgtc ttcagcacgt cgcaatcgcg taccaaacac atcacgcata tgattaattt
     6661 gttcaattgt ataaccaaca cgttgctcaa cccgtcctcg aatttccata tccgggtgcg
     6721 gtagtcgccc tgctttctcg gcatctctga tagcctgaga agaaacccca actaaatccg
     6781 ctgcttcacc tattctccag cgccgggtta ttttcctcgc ttccgggctg tcatcattaa
     6841 actgtgcaat ggcgatagcc ttcgtcattt catgaccagc gtttatgcac tggttaagtg
     6901 tttccatgag tttcattctg aacatccttt aatcattgct ttgcgttttt ttattaaatc
     6961 ttgcaattta ctgcaaagca acaacaaaat cgcaaagtca tcaaaaaacc gcaaagttgt
     7021 ttaaaataag agcaacacta caaaaggaga taagaagagc acatacctca gtcacttatt
     7081 atcactagcg ctcgccgcag ccgtgtaacc gagcatagcg agcgaactgg cgaggaagca
     7141 aagaagaact gttctgtcag atagctctta cgctcagcgc aagaagaaat atccaccgtg
     7201 ggaaaaactc caggtagagg tacacacgcg gatagccaat tcagagtaat aaactgtgat
     7261 aatcaaccct catcaatgat gacgaactaa cccccgatat caagtcacat gacgaaggga
     7321 aagagaagga aatcaactgt gacaaactgc cctcaaattt ggcttcctta aaaattacag
     7381 ttcaaaaagt atgagaaaat ccatgcaggc tgaaggaaac agcaaaactg tgacaaatta
     7441 ccctcagtag gtcagaacaa atgtgacgaa ccaccctcaa atctgtgaca gataaccctc
     7501 agactatcct gtcgtcatgg aagtgatatc gcggaaggaa aatacgatat gagtcgtctg
     7561 gcggcctttc tttttctcaa tgtatgagag gcgcattgga gttctgctgt tgatctcatt
     7621 aacacagacc tgcaggaagc ggcggcggaa gtcaggcata cgctggtaac tttgaggcag
     7681 ctggtaacgc tctatgatcc agtcgatttt cagagagacg atgcctgagc catccggctt
     7741 acgatactga cacagggatt cgta
//
