LOCUS       sequence-436630-        7906 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To track the natural FlhD promoter's transcriptional dynamics.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7906)
  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 7906)
  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: FlhD-GFP
FEATURES             Location/Qualifiers
     source          1..7906
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(839..2014)
                     /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    2340..2590
                     /gene="incC"
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     CDS             complement(2593..3348)
                     /codon_start=1
                     /gene="repE"
                     /product="replication initiation protein for the bacterial 
                     F plasmid"
                     /label=repE
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     rep_origin      3439..3658
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     rep_origin      3726..4257
                     /label=oriV
                     /note="incP origin of replication"
     rep_origin      3734..4348
                     /label=oriV
                     /note="origin of replication for the bacterial F plasmid"
     promoter        5206..5308
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             5309..5968
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(5375..5394)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      complement(6103..6197)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(6260..6973)
                     /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(6300..6320)
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     6915..6943
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     RBS             6979..6987
                     /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      7530..7616
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(join(7774..7906,1..839))
                     /codon_start=1
                     /gene="sopB"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopB
                     /translation="MKRAPVIPKHTLNTQPVEDTSLSTPAAPMVDSLIARVGVMARGNA
                     ITLPVCGRDVKFTLEVLRGDSVEKTSRVWSGNERDQELLTEDALDDLIPSFLLTGQQTP
                     AFGRRVSGVIEIADGSRRRKAAALTESDYRVLVGELDDEQMAALSRLGNDYRPTSAYER
                     GQRYASRLQNEFAGNISALADAENISRKIITRCINTAKLPKSVVALFSHPGELSARSGD
                     ALQKAFTDKEELLKQQASNLHEQKKAGVIFEAEEVITLLTSVLKTSSASRTSLSSRHQF
                     APGATVLYKGDKMVLNLDRSRVPTECIEKIEAILKELEKPAP"
ORIGIN
        1 tgatgtcgtg agcttaaact agttcttgat gcagatgacg ttttaagcac agaagttaaa
       61 agagtgataa cttcttcagc ttcaaatatc accccagctt ttttctgctc atgaaggtta
      121 gatgcctgct gcttaagtaa ttcctcttta tctgtaaagg ctttttgaag tgcatcacct
      181 gaccgggcag atagttcacc ggggtgagaa aaaagagcaa caactgattt aggcaatttg
      241 gcggtgttga tacagcgggt aataatctta cgtgaaatat tttccgcatc agccagcgca
      301 gaaatatttc cagcaaattc attctgcaat cggcttgcat aacgctgacc acgttcataa
      361 gcacttgttg ggcgataatc gttacccaat ctggataatg cagccatctg ctcatcatcc
      421 agctcgccaa ccagaacacg ataatcactt tcggtaagtg cagcagcttt acgacggcga
      481 ctcccatcgg caatttctat gacaccagat actcttcgac cgaacgccgg tgtctgttga
      541 ccagtcagta gaaaagaagg gatgagatca tccagtgcgt cctcagtaag cagctcctgg
      601 tcacgttcat tacctgacca tacccgagag gtcttctcaa cactatcacc ccggagcact
      661 tcaagagtaa acttcacatc ccgaccacat acaggcaaag taatggcatt accgcgagcc
      721 attactccta cgcgcgcaat taacgaatcc accatcgggg cagctggtgt cgataacgaa
      781 gtatcttcaa ccggttgagt attgagcgta tgttttggaa taacaggcgc acgcttcatt
      841 atctaatctc ccagcgtggt ttaatcagac gatcgaaaat ttcattgcag acaggttccc
      901 aaatagaaag agcatttctc caggcaccag ttgaagagcg ttgatcaatg gcctgttcaa
      961 aaacagttct catccggatc tgacctttac caacttcatc cgtttcacgt acaacatttt
     1021 ttagaaccat gcttccccag gcatcccgaa tttgctcctc catccacggg gactgagagc
     1081 cattactatt gctgtatttg gtaagcaaaa tacgtacatc aggctcgaac cctttaagat
     1141 caacgttctt gagcagatca cgaagcatat cgaaaaactg cagtgcggag gtgtagtcaa
     1201 acaactcagc aggcgtggga acaatcagca catcagcagc acatacgaca ttaatcgtgc
     1261 cgatacccag gttaggcgcg ctgtcaataa ctatgacatc atagtcatga gcaacagttt
     1321 caatggccag tcggagcatc aggtgtggat cggtgggcag tttaccttca tcaaatttgc
     1381 ccattaactc agtttcaata cggtgcagag ccagacagga aggaataatg tcaagccccg
     1441 gccagcaagt gggctttatt gcataagtga catcgtcctt ttccccaaga tagaaaggca
     1501 ggagagtgtc ttctgcatga atatgaagat ctggtaccca tccgtgatac attgaggctg
     1561 ttccctgggg gtcgttacct tccacgagca aaacacgtag ccccttcaga gccagatcct
     1621 gagcaagatg aacagaaact gaggttttgt aaacgccacc tttatgggca gcaaccccga
     1681 tcaccggtgg aaatacgtct tcagcacgtc gcaatcgcgt accaaacaca tcacgcatat
     1741 gattaatttg ttcaattgta taaccaacac gttgctcaac ccgtcctcga atttccatat
     1801 ccgggtgcgg tagtcgccct gctttctcgg catctctgat agcctgagaa gaaaccccaa
     1861 ctaaatccgc tgcttcacct attctccagc gccgggttat tttcctcgct tccgggctgt
     1921 catcattaaa ctgtgcaatg gcgatagcct tcgtcatttc atgaccagcg tttatgcact
     1981 ggttaagtgt ttccatgagt ttcattctga acatccttta atcattgctt tgcgtttttt
     2041 tattaaatct tgcaatttac tgcaaagcaa caacaaaatc gcaaagtcat caaaaaaccg
     2101 caaagttgtt taaaataaga gcaacactac aaaaggagat aagaagagca catacctcag
     2161 tcacttatta tcactagcgc tcgccgcagc cgtgtaaccg agcatagcga gcgaactggc
     2221 gaggaagcaa agaagaactg ttctgtcaga tagctcttac gctcagcgca agaagaaata
     2281 tccaccgtgg gaaaaactcc aggtagaggt acacacgcgg atagccaatt cagagtaata
     2341 aactgtgata atcaaccctc atcaatgatg acgaactaac ccccgatatc aagtcacatg
     2401 acgaagggaa agagaaggaa atcaactgtg acaaactgcc ctcaaatttg gcttccttaa
     2461 aaattacagt tcaaaaagta tgagaaaatc catgcaggct gaaggaaaca gcaaaactgt
     2521 gacaaattac cctcagtagg tcagaacaaa tgtgacgaac caccctcaaa tctgtgacag
     2581 ataaccctca gactatcctg tcgtcatgga agtgatatcg cggaaggaaa atacgatatg
     2641 agtcgtctgg cggcctttct ttttctcaat gtatgagagg cgcattggag ttctgctgtt
     2701 gatctcatta acacagacct gcaggaagcg gcggcggaag tcaggcatac gctggtaact
     2761 ttgaggcagc tggtaacgct ctatgatcca gtcgattttc agagagacga tgcctgagcc
     2821 atccggctta cgatactgac acagggattc gtataaacgc atggcatacg gattggtgat
     2881 ttcttttgtt tcactaagcc gaaactgcgt aaaccggttc tgtaacccga taaagaaggg
     2941 aatgagatat gggttgatat gtacactgta aagccctctg gatggactgt gcgcacgttt
     3001 gataaaccaa ggaaaagatt catagccttt ttcatcgccg gcatcctctt cagggcgata
     3061 aaaaaccact tccttccccg cgaaactctt caatgcctgc cgtatatcct tactggcttc
     3121 cgcagaggtc aatccgaata tttcagcata tttagcaaca tggatctcgc agataccgtc
     3181 atgttcctgt agggtgccat cagattttct gatctggtca acgaacagat acagcatacg
     3241 tttttgatcc cgggagagac tatatgccgc ctcagtgagg tcgtttgact ggacgattcg
     3301 cgggctattt ttacgtttct tgtgattgat aaccgctgtt tccgccatga cagatccatg
     3361 tgaagtgtga caagttttta gattgtcaca ctaaataaaa aagagtcaat aagcagggat
     3421 aactttgtga aaaaacagct tcttctgagg gcaatttgtc acagggttaa gggcaatttg
     3481 tcacagacag gactgtcatt tgagggtgat ttgtcacact gaaagggcaa tttgtcacaa
     3541 caccttctct agaaccagca tggataaagg cctacaaggc gctctaaaaa agaagatcta
     3601 aaaactataa aaaaaataat tataaaaata tccccgtgga taagtggata accccaaggg
     3661 aagttttttc aggcatcgtg tgtaagcaga atatataagt gctgttccct ggtgcttcct
     3721 cgctcactcg accgggaggg ttcgagaagg gggggcaccc cccttcggcg tgcgcggtca
     3781 cgcgcacagg gcgcagccct ggttaaaaac aaggtttata aatattggtt taaaagcagg
     3841 ttaaaagaca ggttagcggt ggccgaaaaa cgggcggaaa cccttgcaaa tgctggattt
     3901 tctgcctgtg gacagcccct caaatgtcaa taggtgcgcc cctcatctgt cagcactctg
     3961 cccctcaagt gtcaaggatc gcgcccctca tctgtcagta gtcgcgcccc tcaagtgtca
     4021 ataccgcagg gcacttatcc ccaggcttgt ccacatcatc tgtgggaaac tcgcgtaaaa
     4081 tcaggcgttt tcgccgattt gcgaggctgg ccagctccac gtcgccggcc gaaatcgagc
     4141 ctgcccctca tctgtcaacg ccgcgccggg tgagtcggcc cctcaagtgt caacgtccgc
     4201 ccctcatctg tcagtgaggg ccaagttttc cgcgaggtat ccacaacgcc ggcggccggc
     4261 cgcggtgtct cgcacacggc ttcgacggcg tttctggcgc gtttgcaggg ccatagacgg
     4321 ccgccagccc agcggcgagg gcaaccagcc gagggcttcg ccctgtcgct cgactgcggc
     4381 gagcactact ggctgtaaaa ggacagacca catcatggtt ctgtgttcat taggttgttc
     4441 tgtccattgc tgacataatc cgctccactt caacgtaaca ccgcacgaag atttctattg
     4501 ttcctgaagg catattcaaa tcgttttcgt taccgcttgc tggcatcatg acagaacact
     4561 acttcctata aacgctacac aggctcctga gattaataat gcggatctct acgataatgg
     4621 gagattttcc cgactgtttc gttcgcttct cagtggataa cagccagctt ctctgtttaa
     4681 cagacaaaaa cagcatatcc actcagttcc acatttccat ataaaggcca aggcatttat
     4741 tctcaggata attgtttcag catcgcaacc gcatcagact ccggcatcgc aaactgcacc
     4801 cggtgccggg cagccacatc cagcgcaaaa accttcgtgt agacttccgt tgaactgatg
     4861 gacttatgtc ccatcaggct ttgcagaact ttcagcggta taccggcata cagcatgtgc
     4921 atcgcatagg aatggcggaa cgtatgtggt gtgaccggaa cagagaacgt cacaccgtca
     4981 gcagcagcgg cggcaaccgc ctccccaatc caggtcctga ccgttctgtc cgtcacttcc
     5041 cagatccgcg ctttctctgt ccttcctgtg cgacggttac gccgctccat gagcttatcg
     5101 cgaataaata cctgtgacgg aagatcactt cgcagaataa ataaatcctg gtgtccctgt
     5161 tgataccggg aagccctggg ccaacttttg gcgaaaatga gacgttgatc ggcacgtaag
     5221 aggttccaac tttcaccata atgaaataag atcactaccg ggcgtatttt ttgagttatc
     5281 gagattttca ggagctaagg aagctaaaat ggagaaaaaa atcactggat ataccaccgt
     5341 tgatatatcc caatggcatc gtaaagaaca ttttgaggca tttcagtcag ttgctcaatg
     5401 tacctataac cagaccgttc agctggatat tacggccttt ttaaagaccg taaagaaaaa
     5461 taagcacaag ttttatccgg cctttattca cattcttgcc cgcctgatga atgctcatcc
     5521 ggaatttcgt atggcaatga aagacggtga gctggtgata tgggatagtg ttcacccttg
     5581 ttacaccgtt ttccatgagc aaactgaaac gttttcatcg ctctggagtg aataccacga
     5641 cgatttccgg cagtttctac acatatattc gcaagatgtg gcgtgttacg gtgaaaacct
     5701 ggcctatttc cctaaagggt ttattgagaa tatgtttttc gtctcagcca atccctgggt
     5761 gagtttcacc agttttgatt taaacgtggc caatatggac aacttcttcg cccccgtttt
     5821 cacgatgggc aaatattata cgcaaggcga caaggtgctg atgccgctgg cgattcaggt
     5881 tcatcatgcc gtttgtgatg gcttccatgt cggcagaatg cttaatgaat tacaacagta
     5941 ctgcgatgag tggcagggcg gggcgtaatt tttttaaggc agttattggt gcccttaaac
     6001 gcctggttgc tacgcctgaa taagtgataa taagcggatg aatggcagaa attcgatgat
     6061 aagctgtcaa acatgagaat tggtctcgac gagtatgctt ggattctcac caataaaaaa
     6121 cgcccggcgg caaccgagcg ttctgaacaa atccagatgg agttctgagg tcattactgg
     6181 atctatcaac aggagtccaa gcgacgtctg tgcaagtact actgttctgc agtcacttga
     6241 attcgatacc cagctgttat ttgtatagtt catccatgcc atgtgtaatc ccagcagcgg
     6301 ttacaaactc aagaaggacc atgtggtctc tcttttcgtt gggatctttc gaaagggcag
     6361 attgtgtgga caggtaatgg ttgtctggta aaaggacagg gccatcgcca atcggagtat
     6421 tttgttgata atggtctgct agttgaacgc ttccatcttc aatgttgtgt ctaattttga
     6481 agttaacttt gattccattc ttttgtttgt ctgccatgat gtatacattg tgtgagttat
     6541 agttgtattc caatttgtgt ccaagaatgt ttccatcttc tttaaaatca atacctttta
     6601 actcgattct attaacaagg gtatcacctt caaacttgac ttcagcacgt gtcttgtagt
     6661 tcccgtcatc tttgaaaaat atagttcttt cctgtacgta accttcgggc atggcactct
     6721 tgaaaaagtc atgctgtttc atgtgatctg ggtatctcgc aaagcattga acaccataac
     6781 cgaaagtagt gacaagtgtt ggccacggaa caggtagttt tccagtagtg caaataaatt
     6841 taagggtaag ttttccgtat gttgcatcac cttcaccctc tccactgaca gaaaatttgt
     6901 gcccattaac atcaccatct aattcaacaa gaattgggac aactccagtg aaaagttctt
     6961 ctcctttacg catattatac ctccttaatt tagtgtcatc tacaactggc attattccca
     7021 cccagaataa ccaactttat ttttatgcgg tttcaccgca ccccgtgatg tcgccgggaa
     7081 gccccggtaa aaaataatta gcattagaat agttgcgata agctgcaata agcagaacca
     7141 cctttttggt ttaatatgtc cttacaaata gaaatgggtc tttacactta tctaagattt
     7201 ttcctaaatc gacgcaactg tactcgtcac tacacgcaca tacaacggag gggggctgcg
     7261 attttcaata atgcgtgatg cagatcacac aaaacactca attacttaac ataaatgggt
     7321 aatgactcca acttattgat agtgttttat gttcagataa tgcccgatga ctttgtcatg
     7381 cagctccacc gattttgaga acgacagcga cttccgtccc agccgtgcca ggtgctgcct
     7441 cagattcagg ttatgccgct caattcgctg cgtatatcgc ttcctaggga ccgtctcgag
     7501 agaatcaata ttaatccaac gcgtggcatc aaataaaacg aaaggctcag tcgaaagact
     7561 gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc
     7621 cgccctagac ttaggcgttc ggctgcggcg agcggtatca gctcactcaa agaagtgggg
     7681 cccagagaga atattcaggc cagttatgct ttctggcctg taacaaagga cattaagtaa
     7741 agacagataa acgtagacta aaacgtggtc gcatcagggt gctggctttt caagttcctt
     7801 aagaatggcc tcaattttct ctatacactc agttggaaca cgggacctgt ccaggttaag
     7861 caccatttta tcgcccttat acaatactgt cgctccagga gcaaac
//
