LOCUS       sequence-440606-        8394 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To track the natural Fur promoter's transcriptional dynamics.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8394)
  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 8394)
  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: Fur-GFP
FEATURES             Location/Qualifiers
     source          1..8394
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(940..1026)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     RBS             2057..2065
                     /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             2071..2784
                     /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(2101..2129)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2724..2744
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      2847..2941
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3076..3735)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3650..3669
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(3736..3838)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     rep_origin      4696..5310
                     /label=oriV
                     /note="origin of replication for the bacterial F plasmid"
     rep_origin      4787..5318
                     /label=oriV
                     /note="incP origin of replication"
     rep_origin      5386..5605
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     CDS             5696..6451
                     /codon_start=1
                     /gene="repE"
                     /product="replication initiation protein for the bacterial 
                     F plasmid"
                     /label=repE
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     misc_feature    6454..6704
                     /gene="incC"
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     CDS             7030..8205
                     /codon_start=1
                     /gene="sopA"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopA
                     /translation="MFRMKLMETLNQCINAGHEMTKAIAIAQFNDDSPEARKITRRWRI
                     GEAADLVGVSSQAIRDAEKAGRLPHPDMEIRGRVEQRVGYTIEQINHMRDVFGTRLRRA
                     EDVFPPVIGVAAHKGGVYKTSVSVHLAQDLALKGLRVLLVEGNDPQGTASMYHGWVPDL
                     HIHAEDTLLPFYLGEKDDVTYAIKPTCWPGLDIIPSCLALHRIETELMGKFDEGKLPTD
                     PHLMLRLAIETVAHDYDVIVIDSAPNLGIGTINVVCAADVLIVPTPAELFDYTSALQFF
                     DMLRDLLKNVDLKGFEPDVRILLTKYSNSNGSQSPWMEEQIRDAWGSMVLKNVVRETDE
                     VGKGQIRMRTVFEQAIDQRSSTGAWRNALSIWEPVCNEIFDRLIKPRWEIR"
     CDS             join(8205..8394,1..782)
                     /codon_start=1
                     /gene="sopB"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopB
                     /translation="MKRAPVIPKHTLNTQPVEDTSLSTPAAPMVDSLIARVGVMARGNA
                     ITLPVCGRDVKFTLEVLRGDSVEKTSRVWSGNERDQELLTEDALDDLIPSFLLTGQQTP
                     AFGRRVSGVIEIADGSRRRKAAALTESDYRVLVGELDDEQMAALSRLGNDYRPTSAYER
                     GQRYASRLQNEFAGNISALADAENISRKIITRCINTAKLPKSVVALFSHPGELSARSGD
                     ALQKAFTDKEELLKQQASNLHEQKKAGVIFEAEEVITLLTSVLKTSSASRTSLSSRHQF
                     APGATVLYKGDKMVLNLDRSRVPTECIEKIEAILKELEKPAP"
ORIGIN
        1 gtgatagtgt tgagaagacc tctcgggtat ggtcaggtaa tgaacgtgac caggagctgc
       61 ttactgagga cgcactggat gatctcatcc cttcttttct actgactggt caacagacac
      121 cggcgttcgg tcgaagagta tctggtgtca tagaaattgc cgatgggagt cgccgtcgta
      181 aagctgctgc acttaccgaa agtgattatc gtgttctggt tggcgagctg gatgatgagc
      241 agatggctgc attatccaga ttgggtaacg attatcgccc aacaagtgct tatgaacgtg
      301 gtcagcgtta tgcaagccga ttgcagaatg aatttgctgg aaatatttct gcgctggctg
      361 atgcggaaaa tatttcacgt aagattatta cccgctgtat caacaccgcc aaattgccta
      421 aatcagttgt tgctcttttt tctcaccccg gtgaactatc tgcccggtca ggtgatgcac
      481 ttcaaaaagc ctttacagat aaagaggaat tacttaagca gcaggcatct aaccttcatg
      541 agcagaaaaa agctggggtg atatttgaag ctgaagaagt tatcactctt ttaacttctg
      601 tgcttaaaac gtcatctgca tcaagaacta gtttaagctc acgacatcag tttgctcctg
      661 gagcgacagt attgtataag ggcgataaaa tggtgcttaa cctggacagg tcccgtgttc
      721 caactgagtg tatagagaaa attgaggcca ttcttaagga acttgaaaag ccagcaccct
      781 gatgcgacca cgttttagtc tacgtttatc tgtctttact taatgtcctt tgttacaggc
      841 cagaaagcat aactggcctg aatattctct ctgggcccca cttctttgag tgagctgata
      901 ccgctcgccg cagccgaacg cctaagtcta gggcggcgga tttgtcctac tcaggagagc
      961 gttcaccgac aaacaacaga taaaacgaaa ggcccagtct ttcgactgag cctttcgttt
     1021 tatttgatgc cacgcgttgg attaatattg attctctcga gacggtccct aggatttcca
     1081 ctttcatgta gcacagtgtg cagtcctgct cgtttgctga ttccgcaaga ctgcctgttc
     1141 tgctatgatt gcctttatcc gtgggcaatt ttccaccccc atttcaataa gtttcaagag
     1201 gttatttcac tcatggctat cactggcatc tttttcggca gcgacaccgg taataccgaa
     1261 aatatcgcaa aaatgattca aaaacagctt ggtaaagacg ttgccgatgt ccatgacatt
     1321 gcaaaaagca gcaaagaaga tctggaagct tatgacattc tgctgctggg catcccaacc
     1381 tggtattacg gcgaagcgca gtgtgactgg gatgacttct tcccgactct cgaagagatt
     1441 gatttcaacg gcaaactggt tgcgctgttt ggttgtggtg accaggaaga ttacgccgaa
     1501 tatttctgcg acgcattggg caccatccgc gacatcattg aaccgcgcgg tgcaaccatc
     1561 gttggtcact ggccaactgc gggctatcat ttcgaagcat caaaaggtct ggcagatgac
     1621 gaccactttg tcggtctggc tatcgacgaa gaccgtcagc cggaactgac cgctgaacgt
     1681 gtagaaaaat gggttaaaca gatttctgaa gagttgcatc tcgacgaaat tctcaatgcc
     1741 tgatgtgatg cggcgtagac tcatgtctac gccgtattaa tagataatgc caatcaaaat
     1801 aattgctaca aatttgtaac ttttgctgtt gtacctgtac aatgtcccgg tgttcaagtg
     1861 gccttgccgt tgtaaatgta agctgtgcca cgtttttatt aacaatattt gccagggact
     1921 tgtggttttc atttaggcgt ggcaattcta taatgatacg cattatctca agagcaaatt
     1981 ctgtcacttc ttctaatgaa gtgaaccgct tagtaacagg acagattccg catgccagtt
     2041 gtagatgaca ctaaattaag gaggtataat atgcgtaaag gagaagaact tttcactgga
     2101 gttgtcccaa ttcttgttga attagatggt gatgttaatg ggcacaaatt ttctgtcagt
     2161 ggagagggtg aaggtgatgc aacatacgga aaacttaccc ttaaatttat ttgcactact
     2221 ggaaaactac ctgttccgtg gccaacactt gtcactactt tcggttatgg tgttcaatgc
     2281 tttgcgagat acccagatca catgaaacag catgactttt tcaagagtgc catgcccgaa
     2341 ggttacgtac aggaaagaac tatatttttc aaagatgacg ggaactacaa gacacgtgct
     2401 gaagtcaagt ttgaaggtga tacccttgtt aatagaatcg agttaaaagg tattgatttt
     2461 aaagaagatg gaaacattct tggacacaaa ttggaataca actataactc acacaatgta
     2521 tacatcatgg cagacaaaca aaagaatgga atcaaagtta acttcaaaat tagacacaac
     2581 attgaagatg gaagcgttca actagcagac cattatcaac aaaatactcc gattggcgat
     2641 ggccctgtcc ttttaccaga caaccattac ctgtccacac aatctgccct ttcgaaagat
     2701 cccaacgaaa agagagacca catggtcctt cttgagtttg taaccgctgc tgggattaca
     2761 catggcatgg atgaactata caaataacag ctgggtatcg aattcaagtg actgcagaac
     2821 agtagtactt gcacagacgt cgcttggact cctgttgata gatccagtaa tgacctcaga
     2881 actccatctg gatttgttca gaacgctcgg ttgccgccgg gcgtttttta ttggtgagaa
     2941 tccaagcata ctcgtcgaga ccaattctca tgtttgacag cttatcatcg aatttctgcc
     3001 attcatccgc ttattatcac ttattcaggc gtagcaacca ggcgtttaag ggcaccaata
     3061 actgccttaa aaaaattacg ccccgccctg ccactcatcg cagtactgtt gtaattcatt
     3121 aagcattctg ccgacatgga agccatcaca aacggcatga tgaacctgaa tcgccagcgg
     3181 catcagcacc ttgtcgcctt gcgtataata tttgcccatc gtgaaaacgg gggcgaagaa
     3241 gttgtccata ttggccacgt ttaaatcaaa actggtgaaa ctcacccagg gattggctga
     3301 gacgaaaaac atattctcaa taaacccttt agggaaatag gccaggtttt caccgtaaca
     3361 cgccacatct tgcgaatata tgtgtagaaa ctgccggaaa tcgtcgtggt attcactcca
     3421 gagcgatgaa aacgtttcag tttgctcatg gaaaacggtg taacaagggt gaacactatc
     3481 ccatatcacc agctcaccgt ctttcattgc catacgaaat tccggatgag cattcatcag
     3541 gcgggcaaga atgtgaataa aggccggata aaacttgtgc ttatttttct ttacggtctt
     3601 taaaaaggcc gtaatatcca gctgaacggt ctggttatag gtacattgag caactgactg
     3661 aaatgcctca aaatgttctt tacgatgcca ttgggatata tcaacggtgg tatatccagt
     3721 gatttttttc tccattttag cttccttagc tcctgaaaat ctcgataact caaaaaatac
     3781 gcccggtagt gatcttattt cattatggtg aaagttggaa cctcttacgt gccgatcaac
     3841 gtctcatttt cgccaaaagt tggcccaggg cttcccggta tcaacaggga caccaggatt
     3901 tatttattct gcgaagtgat cttccgtcac aggtatttat tcgcgataag ctcatggagc
     3961 ggcgtaaccg tcgcacagga aggacagaga aagcgcggat ctgggaagtg acggacagaa
     4021 cggtcaggac ctggattggg gaggcggttg ccgccgctgc tgctgacggt gtgacgttct
     4081 ctgttccggt cacaccacat acgttccgcc attcctatgc gatgcacatg ctgtatgccg
     4141 gtataccgct gaaagttctg caaagcctga tgggacataa gtccatcagt tcaacggaag
     4201 tctacacgaa ggtttttgcg ctggatgtgg ctgcccggca ccgggtgcag tttgcgatgc
     4261 cggagtctga tgcggttgcg atgctgaaac aattatcctg agaataaatg ccttggcctt
     4321 tatatggaaa tgtggaactg agtggatatg ctgtttttgt ctgttaaaca gagaagctgg
     4381 ctgttatcca ctgagaagcg aacgaaacag tcgggaaaat ctcccattat cgtagagatc
     4441 cgcattatta atctcaggag cctgtgtagc gtttatagga agtagtgttc tgtcatgatg
     4501 ccagcaagcg gtaacgaaaa cgatttgaat atgccttcag gaacaataga aatcttcgtg
     4561 cggtgttacg ttgaagtgga gcggattatg tcagcaatgg acagaacaac ctaatgaaca
     4621 cagaaccatg atgtggtctg tccttttaca gccagtagtg ctcgccgcag tcgagcgaca
     4681 gggcgaagcc ctcggctggt tgccctcgcc gctgggctgg cggccgtcta tggccctgca
     4741 aacgcgccag aaacgccgtc gaagccgtgt gcgagacacc gcggccggcc gccggcgttg
     4801 tggatacctc gcggaaaact tggccctcac tgacagatga ggggcggacg ttgacacttg
     4861 aggggccgac tcacccggcg cggcgttgac agatgagggg caggctcgat ttcggccggc
     4921 gacgtggagc tggccagcct cgcaaatcgg cgaaaacgcc tgattttacg cgagtttccc
     4981 acagatgatg tggacaagcc tggggataag tgccctgcgg tattgacact tgaggggcgc
     5041 gactactgac agatgagggg cgcgatcctt gacacttgag gggcagagtg ctgacagatg
     5101 aggggcgcac ctattgacat ttgaggggct gtccacaggc agaaaatcca gcatttgcaa
     5161 gggtttccgc ccgtttttcg gccaccgcta acctgtcttt taacctgctt ttaaaccaat
     5221 atttataaac cttgttttta accagggctg cgccctgtgc gcgtgaccgc gcacgccgaa
     5281 ggggggtgcc cccccttctc gaaccctccc ggtcgagtga gcgaggaagc accagggaac
     5341 agcacttata tattctgctt acacacgatg cctgaaaaaa cttcccttgg ggttatccac
     5401 ttatccacgg ggatattttt ataattattt tttttatagt ttttagatct tcttttttag
     5461 agcgccttgt aggcctttat ccatgctggt tctagagaag gtgttgtgac aaattgccct
     5521 ttcagtgtga caaatcaccc tcaaatgaca gtcctgtctg tgacaaattg cccttaaccc
     5581 tgtgacaaat tgccctcaga agaagctgtt ttttcacaaa gttatccctg cttattgact
     5641 cttttttatt tagtgtgaca atctaaaaac ttgtcacact tcacatggat ctgtcatggc
     5701 ggaaacagcg gttatcaatc acaagaaacg taaaaatagc ccgcgaatcg tccagtcaaa
     5761 cgacctcact gaggcggcat atagtctctc ccgggatcaa aaacgtatgc tgtatctgtt
     5821 cgttgaccag atcagaaaat ctgatggcac cctacaggaa catgacggta tctgcgagat
     5881 ccatgttgct aaatatgctg aaatattcgg attgacctct gcggaagcca gtaaggatat
     5941 acggcaggca ttgaagagtt tcgcggggaa ggaagtggtt ttttatcgcc ctgaagagga
     6001 tgccggcgat gaaaaaggct atgaatcttt tccttggttt atcaaacgtg cgcacagtcc
     6061 atccagaggg ctttacagtg tacatatcaa cccatatctc attcccttct ttatcgggtt
     6121 acagaaccgg tttacgcagt ttcggcttag tgaaacaaaa gaaatcacca atccgtatgc
     6181 catgcgttta tacgaatccc tgtgtcagta tcgtaagccg gatggctcag gcatcgtctc
     6241 tctgaaaatc gactggatca tagagcgtta ccagctgcct caaagttacc agcgtatgcc
     6301 tgacttccgc cgccgcttcc tgcaggtctg tgttaatgag atcaacagca gaactccaat
     6361 gcgcctctca tacattgaga aaaagaaagg ccgccagacg actcatatcg tattttcctt
     6421 ccgcgatatc acttccatga cgacaggata gtctgagggt tatctgtcac agatttgagg
     6481 gtggttcgtc acatttgttc tgacctactg agggtaattt gtcacagttt tgctgtttcc
     6541 ttcagcctgc atggattttc tcatactttt tgaactgtaa tttttaagga agccaaattt
     6601 gagggcagtt tgtcacagtt gatttccttc tctttccctt cgtcatgtga cttgatatcg
     6661 ggggttagtt cgtcatcatt gatgagggtt gattatcaca gtttattact ctgaattggc
     6721 tatccgcgtg tgtacctcta cctggagttt ttcccacggt ggatatttct tcttgcgctg
     6781 agcgtaagag ctatctgaca gaacagttct tctttgcttc ctcgccagtt cgctcgctat
     6841 gctcggttac acggctgcgg cgagcgctag tgataataag tgactgaggt atgtgctctt
     6901 cttatctcct tttgtagtgt tgctcttatt ttaaacaact ttgcggtttt ttgatgactt
     6961 tgcgattttg ttgttgcttt gcagtaaatt gcaagattta ataaaaaaac gcaaagcaat
     7021 gattaaagga tgttcagaat gaaactcatg gaaacactta accagtgcat aaacgctggt
     7081 catgaaatga cgaaggctat cgccattgca cagtttaatg atgacagccc ggaagcgagg
     7141 aaaataaccc ggcgctggag aataggtgaa gcagcggatt tagttggggt ttcttctcag
     7201 gctatcagag atgccgagaa agcagggcga ctaccgcacc cggatatgga aattcgagga
     7261 cgggttgagc aacgtgttgg ttatacaatt gaacaaatta atcatatgcg tgatgtgttt
     7321 ggtacgcgat tgcgacgtgc tgaagacgta tttccaccgg tgatcggggt tgctgcccat
     7381 aaaggtggcg tttacaaaac ctcagtttct gttcatcttg ctcaggatct ggctctgaag
     7441 gggctacgtg ttttgctcgt ggaaggtaac gacccccagg gaacagcctc aatgtatcac
     7501 ggatgggtac cagatcttca tattcatgca gaagacactc tcctgccttt ctatcttggg
     7561 gaaaaggacg atgtcactta tgcaataaag cccacttgct ggccggggct tgacattatt
     7621 ccttcctgtc tggctctgca ccgtattgaa actgagttaa tgggcaaatt tgatgaaggt
     7681 aaactgccca ccgatccaca cctgatgctc cgactggcca ttgaaactgt tgctcatgac
     7741 tatgatgtca tagttattga cagcgcgcct aacctgggta tcggcacgat taatgtcgta
     7801 tgtgctgctg atgtgctgat tgttcccacg cctgctgagt tgtttgacta cacctccgca
     7861 ctgcagtttt tcgatatgct tcgtgatctg ctcaagaacg ttgatcttaa agggttcgag
     7921 cctgatgtac gtattttgct taccaaatac agcaatagta atggctctca gtccccgtgg
     7981 atggaggagc aaattcggga tgcctgggga agcatggttc taaaaaatgt tgtacgtgaa
     8041 acggatgaag ttggtaaagg tcagatccgg atgagaactg tttttgaaca ggccattgat
     8101 caacgctctt caactggtgc ctggagaaat gctctttcta tttgggaacc tgtctgcaat
     8161 gaaattttcg atcgtctgat taaaccacgc tgggagatta gataatgaag cgtgcgcctg
     8221 ttattccaaa acatacgctc aatactcaac cggttgaaga tacttcgtta tcgacaccag
     8281 ctgccccgat ggtggattcg ttaattgcgc gcgtaggagt aatggctcgc ggtaatgcca
     8341 ttactttgcc tgtatgtggt cgggatgtga agtttactct tgaagtgctc cggg
//
