LOCUS       sequence-436629-        7630 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To track the natural Fnr promoter's transcriptional dynamics.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7630)
  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 7630)
  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: Fnr-GFP
FEATURES             Location/Qualifiers
     source          1..7630
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(455..541)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     RBS             808..816
                     /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             822..1535
                     /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(852..880)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1475..1495
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      1598..1692
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1827..2486)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2401..2420
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2487..2589)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     rep_origin      3447..4061
                     /label=oriV
                     /note="origin of replication for the bacterial F plasmid"
     rep_origin      3538..4069
                     /label=oriV
                     /note="incP origin of replication"
     rep_origin      4137..4356
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     CDS             4447..5202
                     /codon_start=1
                     /gene="repE"
                     /product="replication initiation protein for the bacterial 
                     F plasmid"
                     /label=repE
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     misc_feature    5205..5455
                     /gene="incC"
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     CDS             5781..6956
                     /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(6956..7630,1..297)
                     /codon_start=1
                     /gene="sopB"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopB
                     /translation="MKRAPVIPKHTLNTQPVEDTSLSTPAAPMVDSLIARVGVMARGNA
                     ITLPVCGRDVKFTLEVLRGDSVEKTSRVWSGNERDQELLTEDALDDLIPSFLLTGQQTP
                     AFGRRVSGVIEIADGSRRRKAAALTESDYRVLVGELDDEQMAALSRLGNDYRPTSAYER
                     GQRYASRLQNEFAGNISALADAENISRKIITRCINTAKLPKSVVALFSHPGELSARSGD
                     ALQKAFTDKEELLKQQASNLHEQKKAGVIFEAEEVITLLTSVLKTSSASRTSLSSRHQF
                     APGATVLYKGDKMVLNLDRSRVPTECIEKIEAILKELEKPAP"
ORIGIN
        1 aaagccttta cagataaaga ggaattactt aagcagcagg catctaacct tcatgagcag
       61 aaaaaagctg gggtgatatt tgaagctgaa gaagttatca ctcttttaac ttctgtgctt
      121 aaaacgtcat ctgcatcaag aactagttta agctcacgac atcagtttgc tcctggagcg
      181 acagtattgt ataagggcga taaaatggtg cttaacctgg acaggtcccg tgttccaact
      241 gagtgtatag agaaaattga ggccattctt aaggaacttg aaaagccagc accctgatgc
      301 gaccacgttt tagtctacgt ttatctgtct ttacttaatg tcctttgtta caggccagaa
      361 agcataactg gcctgaatat tctctctggg ccccacttct ttgagtgagc tgataccgct
      421 cgccgcagcc gaacgcctaa gtctagggcg gcggatttgt cctactcagg agagcgttca
      481 ccgacaaaca acagataaaa cgaaaggccc agtctttcga ctgagccttt cgttttattt
      541 gatgccacgc gttggattaa tattgattct ctcgagacgg tccctaggac attaaacaat
      601 ttgtgccagc ttgttcacac ttttatgtaa agttaccctt aacaacttaa gggttttcaa
      661 atagatagac atatatttac atctaatatc ggaattctct gctgttaagg tttgcttaga
      721 cttacttgct ccctaaaaag atgttaaaat tgacaaatat caattacggc ttgagcagac
      781 ctatgccagt tgtagatgac actaaattaa ggaggtataa tatgcgtaaa ggagaagaac
      841 ttttcactgg agttgtccca attcttgttg aattagatgg tgatgttaat gggcacaaat
      901 tttctgtcag tggagagggt gaaggtgatg caacatacgg aaaacttacc cttaaattta
      961 tttgcactac tggaaaacta cctgttccgt ggccaacact tgtcactact ttcggttatg
     1021 gtgttcaatg ctttgcgaga tacccagatc acatgaaaca gcatgacttt ttcaagagtg
     1081 ccatgcccga aggttacgta caggaaagaa ctatattttt caaagatgac gggaactaca
     1141 agacacgtgc tgaagtcaag tttgaaggtg atacccttgt taatagaatc gagttaaaag
     1201 gtattgattt taaagaagat ggaaacattc ttggacacaa attggaatac aactataact
     1261 cacacaatgt atacatcatg gcagacaaac aaaagaatgg aatcaaagtt aacttcaaaa
     1321 ttagacacaa cattgaagat ggaagcgttc aactagcaga ccattatcaa caaaatactc
     1381 cgattggcga tggccctgtc cttttaccag acaaccatta cctgtccaca caatctgccc
     1441 tttcgaaaga tcccaacgaa aagagagacc acatggtcct tcttgagttt gtaaccgctg
     1501 ctgggattac acatggcatg gatgaactat acaaataaca gctgggtatc gaattcaagt
     1561 gactgcagaa cagtagtact tgcacagacg tcgcttggac tcctgttgat agatccagta
     1621 atgacctcag aactccatct ggatttgttc agaacgctcg gttgccgccg ggcgtttttt
     1681 attggtgaga atccaagcat actcgtcgag accaattctc atgtttgaca gcttatcatc
     1741 gaatttctgc cattcatccg cttattatca cttattcagg cgtagcaacc aggcgtttaa
     1801 gggcaccaat aactgcctta aaaaaattac gccccgccct gccactcatc gcagtactgt
     1861 tgtaattcat taagcattct gccgacatgg aagccatcac aaacggcatg atgaacctga
     1921 atcgccagcg gcatcagcac cttgtcgcct tgcgtataat atttgcccat cgtgaaaacg
     1981 ggggcgaaga agttgtccat attggccacg tttaaatcaa aactggtgaa actcacccag
     2041 ggattggctg agacgaaaaa catattctca ataaaccctt tagggaaata ggccaggttt
     2101 tcaccgtaac acgccacatc ttgcgaatat atgtgtagaa actgccggaa atcgtcgtgg
     2161 tattcactcc agagcgatga aaacgtttca gtttgctcat ggaaaacggt gtaacaaggg
     2221 tgaacactat cccatatcac cagctcaccg tctttcattg ccatacgaaa ttccggatga
     2281 gcattcatca ggcgggcaag aatgtgaata aaggccggat aaaacttgtg cttatttttc
     2341 tttacggtct ttaaaaaggc cgtaatatcc agctgaacgg tctggttata ggtacattga
     2401 gcaactgact gaaatgcctc aaaatgttct ttacgatgcc attgggatat atcaacggtg
     2461 gtatatccag tgattttttt ctccatttta gcttccttag ctcctgaaaa tctcgataac
     2521 tcaaaaaata cgcccggtag tgatcttatt tcattatggt gaaagttgga acctcttacg
     2581 tgccgatcaa cgtctcattt tcgccaaaag ttggcccagg gcttcccggt atcaacaggg
     2641 acaccaggat ttatttattc tgcgaagtga tcttccgtca caggtattta ttcgcgataa
     2701 gctcatggag cggcgtaacc gtcgcacagg aaggacagag aaagcgcgga tctgggaagt
     2761 gacggacaga acggtcagga cctggattgg ggaggcggtt gccgccgctg ctgctgacgg
     2821 tgtgacgttc tctgttccgg tcacaccaca tacgttccgc cattcctatg cgatgcacat
     2881 gctgtatgcc ggtataccgc tgaaagttct gcaaagcctg atgggacata agtccatcag
     2941 ttcaacggaa gtctacacga aggtttttgc gctggatgtg gctgcccggc accgggtgca
     3001 gtttgcgatg ccggagtctg atgcggttgc gatgctgaaa caattatcct gagaataaat
     3061 gccttggcct ttatatggaa atgtggaact gagtggatat gctgtttttg tctgttaaac
     3121 agagaagctg gctgttatcc actgagaagc gaacgaaaca gtcgggaaaa tctcccatta
     3181 tcgtagagat ccgcattatt aatctcagga gcctgtgtag cgtttatagg aagtagtgtt
     3241 ctgtcatgat gccagcaagc ggtaacgaaa acgatttgaa tatgccttca ggaacaatag
     3301 aaatcttcgt gcggtgttac gttgaagtgg agcggattat gtcagcaatg gacagaacaa
     3361 cctaatgaac acagaaccat gatgtggtct gtccttttac agccagtagt gctcgccgca
     3421 gtcgagcgac agggcgaagc cctcggctgg ttgccctcgc cgctgggctg gcggccgtct
     3481 atggccctgc aaacgcgcca gaaacgccgt cgaagccgtg tgcgagacac cgcggccggc
     3541 cgccggcgtt gtggatacct cgcggaaaac ttggccctca ctgacagatg aggggcggac
     3601 gttgacactt gaggggccga ctcacccggc gcggcgttga cagatgaggg gcaggctcga
     3661 tttcggccgg cgacgtggag ctggccagcc tcgcaaatcg gcgaaaacgc ctgattttac
     3721 gcgagtttcc cacagatgat gtggacaagc ctggggataa gtgccctgcg gtattgacac
     3781 ttgaggggcg cgactactga cagatgaggg gcgcgatcct tgacacttga ggggcagagt
     3841 gctgacagat gaggggcgca cctattgaca tttgaggggc tgtccacagg cagaaaatcc
     3901 agcatttgca agggtttccg cccgtttttc ggccaccgct aacctgtctt ttaacctgct
     3961 tttaaaccaa tatttataaa ccttgttttt aaccagggct gcgccctgtg cgcgtgaccg
     4021 cgcacgccga aggggggtgc ccccccttct cgaaccctcc cggtcgagtg agcgaggaag
     4081 caccagggaa cagcacttat atattctgct tacacacgat gcctgaaaaa acttcccttg
     4141 gggttatcca cttatccacg gggatatttt tataattatt ttttttatag tttttagatc
     4201 ttctttttta gagcgccttg taggccttta tccatgctgg ttctagagaa ggtgttgtga
     4261 caaattgccc tttcagtgtg acaaatcacc ctcaaatgac agtcctgtct gtgacaaatt
     4321 gcccttaacc ctgtgacaaa ttgccctcag aagaagctgt tttttcacaa agttatccct
     4381 gcttattgac tcttttttat ttagtgtgac aatctaaaaa cttgtcacac ttcacatgga
     4441 tctgtcatgg cggaaacagc ggttatcaat cacaagaaac gtaaaaatag cccgcgaatc
     4501 gtccagtcaa acgacctcac tgaggcggca tatagtctct cccgggatca aaaacgtatg
     4561 ctgtatctgt tcgttgacca gatcagaaaa tctgatggca ccctacagga acatgacggt
     4621 atctgcgaga tccatgttgc taaatatgct gaaatattcg gattgacctc tgcggaagcc
     4681 agtaaggata tacggcaggc attgaagagt ttcgcgggga aggaagtggt tttttatcgc
     4741 cctgaagagg atgccggcga tgaaaaaggc tatgaatctt ttccttggtt tatcaaacgt
     4801 gcgcacagtc catccagagg gctttacagt gtacatatca acccatatct cattcccttc
     4861 tttatcgggt tacagaaccg gtttacgcag tttcggctta gtgaaacaaa agaaatcacc
     4921 aatccgtatg ccatgcgttt atacgaatcc ctgtgtcagt atcgtaagcc ggatggctca
     4981 ggcatcgtct ctctgaaaat cgactggatc atagagcgtt accagttgcc tcaaagttac
     5041 cagcgtatgc ctgacttccg ccgccgcttc ctgcaggtct gtgttaatga gatcaacagc
     5101 agaactccaa tgcgcctctc atacattgag aaaaagaaag gccgccagac gactcatatc
     5161 gtattttcct tccgcgatat cacttccatg acgacaggat agtctgaggg ttatctgtca
     5221 cagatttgag ggtggttcgt cacatttgtt ctgacctact gagggtaatt tgtcacagtt
     5281 ttgctgtttc cttcagcctg catggatttt ctcatacttt ttgaactgta atttttaagg
     5341 aagccaaatt tgagggcagt ttgtcacagt tgatttcctt ctctttccct tcgtcatgtg
     5401 acttgatatc gggggttagt tcgtcatcat tgatgagggt tgattatcac agtttattac
     5461 tctgaattgg ctatccgcgt gtgtacctct acctggagtt tttcccacgg tggatatttc
     5521 ttcttgcgct gagcgtaaga gctatctgac agaacagttc ttctttgctt cctcgccagt
     5581 tcgctcgcta tgctcggtta cacggctgcg gcgagcgcta gtgataataa gtgactgagg
     5641 tatgtgctct tcttatctcc ttttgtagtg ttgctcttat tttaaacaac tttgcggttt
     5701 tttgatgact ttgcgatttt gttgttgctt tgcagtaaat tgcaagattt aataaaaaaa
     5761 cgcaaagcaa tgattaaagg atgttcagaa tgaaactcat ggaaacactt aaccagtgca
     5821 taaacgctgg tcatgaaatg acgaaggcta tcgccattgc acagtttaat gatgacagcc
     5881 cggaagcgag gaaaataacc cggcgctgga gaataggtga agcagcggat ttagttgggg
     5941 tttcttctca ggctatcaga gatgccgaga aagcagggcg actaccgcac ccggatatgg
     6001 aaattcgagg acgggttgag caacgtgttg gttatacaat tgaacaaatt aatcatatgc
     6061 gtgatgtgtt tggtacgcga ttgcgacgtg ctgaagacgt atttccaccg gtgatcgggg
     6121 ttgctgccca taaaggtggc gtttacaaaa cctcagtttc tgttcatctt gctcaggatc
     6181 tggctctgaa ggggctacgt gttttgctcg tggaaggtaa cgacccccag ggaacagcct
     6241 caatgtatca cggatgggta ccagatcttc atattcatgc agaagacact ctcctgcctt
     6301 tctatcttgg ggaaaaggac gatgtcactt atgcaataaa gcccacttgc tggccggggc
     6361 ttgacattat tccttcctgt ctggctctgc accgtattga aactgagtta atgggcaaat
     6421 ttgatgaagg taaactgccc accgatccac acctgatgct ccgactggcc attgaaactg
     6481 ttgctcatga ctatgatgtc atagttattg acagcgcgcc taacctgggt atcggcacga
     6541 ttaatgtcgt atgtgctgct gatgtgctga ttgttcccac gcctgctgag ttgtttgact
     6601 acacctccgc actgcagttt ttcgatatgc ttcgtgatct gctcaagaac gttgatctta
     6661 aagggttcga gcctgatgta cgtattttgc ttaccaaata cagcaatagt aatggctctc
     6721 agtccccgtg gatggaggag caaattcggg atgcctgggg aagcatggtt ctaaaaaatg
     6781 ttgtacgtga aacggatgaa gttggtaaag gtcagatccg gatgagaact gtttttgaac
     6841 aggccattga tcaacgctct tcaactggtg cctggagaaa tgctctttct atttgggaac
     6901 ctgtctgcaa tgaaattttc gatcgtctga ttaaaccacg ctgggagatt agataatgaa
     6961 gcgtgcgcct gttattccaa aacatacgct caatactcaa ccggttgaag atacttcgtt
     7021 atcgacacca gctgccccga tggtggattc gttaattgcg cgcgtaggag taatggctcg
     7081 cggtaatgcc attactttgc ctgtatgtgg tcgggatgtg aagtttactc ttgaagtgct
     7141 ccggggtgat agtgttgaga agacctctcg ggtatggtca ggtaatgaac gtgaccagga
     7201 gctgcttact gaggacgcac tggatgatct catcccttct tttctactga ctggtcaaca
     7261 gacaccggcg ttcggtcgaa gagtatctgg tgtcatagaa attgccgatg ggagtcgccg
     7321 tcgtaaagct gctgcactta ccgaaagtga ttatcgtgtt ctggttggcg agctggatga
     7381 tgagcagatg gctgcattat ccagattggg taacgattat cgcccaacaa gtgcttatga
     7441 acgtggtcag cgttatgcaa gccgattgca gaatgaattt gctggaaata tttctgcgct
     7501 ggctgatgcg gaaaatattt cacgtaagat tattacccgc tgtatcaaca ccgccaaatt
     7561 gcctaaatca gttgttgctc ttttttctca ccccggtgaa ctatctgccc ggtcaggtga
     7621 tgcacttcaa
//
