LOCUS       sequence-466140-        7755 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To track the natural NarP promoter's transcriptional dynamics.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7755)
  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 7755)
  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: NarP-GFP
FEATURES             Location/Qualifiers
     source          1..7755
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      819..1038
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     rep_origin      1106..1637
                     /label=oriV
                     /note="incP origin of replication"
     rep_origin      1114..1728
                     /label=oriV
                     /note="origin of replication for the bacterial F plasmid"
     promoter        2586..2688
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             2689..3348
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2755..2774)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      complement(3483..3577)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3640..4353)
                     /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(3680..3700)
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     4295..4323
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     RBS             4359..4367
                     /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      4759..4845
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(5003..5974)
                     /codon_start=1
                     /gene="sopB"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopB
                     /translation="MKRAPVIPKHTLNTQPVEDTSLSTPAAPMVDSLIARVGVMARGNA
                     ITLPVCGRDVKFTLEVLRGDSVEKTSRVWSGNERDQELLTEDALDDLIPSFLLTGQQTP
                     AFGRRVSGVIEIADGSRRRKAAALTESDYRVLVGELDDEQMAALSRLGNDYRPTSAYER
                     GQRYASRLQNEFAGNISALADAENISRKIITRCINTAKLPKSVVALFSHPGELSARSGD
                     ALQKAFTDKEELLKQQASNLHEQKKAGVIFEAEEVITLLTSVLKTSSASRTSLSSRHQF
                     APGATVLYKGDKMVLNLDRSRVPTECIEKIEAILKELEKPAP"
     CDS             complement(5974..7149)
                     /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    7475..7725
                     /gene="incC"
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     CDS             complement(join(7728..7755,1..728))
                     /codon_start=1
                     /gene="repE"
                     /product="replication initiation protein for the bacterial 
                     F plasmid"
                     /label=repE
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
ORIGIN
        1 cggaaggaaa atacgatatg agtcgtctgg cggcctttct ttttctcaat gtatgagagg
       61 cgcattggag ttctgctgtt gatctcatta acacagacct gcaggaagcg gcggcggaag
      121 tcaggcatac gctggtaact ttgaggcagc tggtaacgct ctatgatcca gtcgattttc
      181 agagagacga tgcctgagcc atccggctta cgatactgac acagggattc gtataaacgc
      241 atggcatacg gattggtgat ttcttttgtt tcactaagcc gaaactgcgt aaaccggttc
      301 tgtaacccga taaagaaggg aatgagatat gggttgatat gtacactgta aagccctctg
      361 gatggactgt gcgcacgttt gataaaccaa ggaaaagatt catagccttt ttcatcgccg
      421 gcatcctctt cagggcgata aaaaaccact tccttccccg cgaaactctt caatgcctgc
      481 cgtatatcct tactggcttc cgcagaggtc aatccgaata tttcagcata tttagcaaca
      541 tggatctcgc agataccgtc atgttcctgt agggtgccat cagattttct gatctggtca
      601 acgaacagat acagcatacg tttttgatcc cgggagagac tatatgccgc ctcagtgagg
      661 tcgtttgact ggacgattcg cgggctattt ttacgtttct tgtgattgat aaccgctgtt
      721 tccgccatga cagatccatg tgaagtgtga caagttttta gattgtcaca ctaaataaaa
      781 aagagtcaat aagcagggat aactttgtga aaaaacagct tcttctgagg gcaatttgtc
      841 acagggttaa gggcaatttg tcacagacag gactgtcatt tgagggtgat ttgtcacact
      901 gaaagggcaa tttgtcacaa caccttctct agaaccagca tggataaagg cctacaaggc
      961 gctctaaaaa agaagatcta aaaactataa aaaaaataat tataaaaata tccccgtgga
     1021 taagtggata accccaaggg aagttttttc aggcatcgtg tgtaagcaga atatataagt
     1081 gctgttccct ggtgcttcct cgctcactcg accgggaggg ttcgagaagg gggggcaccc
     1141 cccttcggcg tgcgcggtca cgcgcacagg gcgcagccct ggttaaaaac aaggtttata
     1201 aatattggtt taaaagcagg ttaaaagaca ggttagcggt ggccgaaaaa cgggcggaaa
     1261 cccttgcaaa tgctggattt tctgcctgtg gacagcccct caaatgtcaa taggtgcgcc
     1321 cctcatctgt cagcactctg cccctcaagt gtcaaggatc gcgcccctca tctgtcagta
     1381 gtcgcgcccc tcaagtgtca ataccgcagg gcacttatcc ccaggcttgt ccacatcatc
     1441 tgtgggaaac tcgcgtaaaa tcaggcgttt tcgccgattt gcgaggctgg ccagctccac
     1501 gtcgccggcc gaaatcgagc ctgcccctca tctgtcaacg ccgcgccggg tgagtcggcc
     1561 cctcaagtgt caacgtccgc ccctcatctg tcagtgaggg ccaagttttc cgcgaggtat
     1621 ccacaacgcc ggcggccggc cgcggtgtct cgcacacggc ttcgacggcg tttctggcgc
     1681 gtttgcaggg ccatagacgg ccgccagccc agcggcgagg gcaaccagcc gagggcttcg
     1741 ccctgtcgct cgactgcggc gagcactact ggctgtaaaa ggacagacca catcatggtt
     1801 ctgtgttcat taggttgttc tgtccattgc tgacataatc cgctccactt caacgtaaca
     1861 ccgcacgaag atttctattg ttcctgaagg catattcaaa tcgttttcgt taccgcttgc
     1921 tggcatcatg acagaacact acttcctata aacgctacac aggctcctga gattaataat
     1981 gcggatctct acgataatgg gagattttcc cgactgtttc gttcgcttct cagtggataa
     2041 cagccagctt ctctgtttaa cagacaaaaa cagcatatcc actcagttcc acatttccat
     2101 ataaaggcca aggcatttat tctcaggata attgtttcag catcgcaacc gcatcagact
     2161 ccggcatcgc aaactgcacc cggtgccggg cagccacatc cagcgcaaaa accttcgtgt
     2221 agacttccgt tgaactgatg gacttatgtc ccatcaggct ttgcagaact ttcagcggta
     2281 taccggcata cagcatgtgc atcgcatagg aatggcggaa cgtatgtggt gtgaccggaa
     2341 cagagaacgt cacaccgtca gcagcagcgg cggcaaccgc ctccccaatc caggtcctga
     2401 ccgttctgtc cgtcacttcc cagatccgcg ctttctctgt ccttcctgtg cgacggttac
     2461 gccgctccat gagcttatcg cgaataaata cctgtgacgg aagatcactt cgcagaataa
     2521 ataaatcctg gtgtccctgt tgataccggg aagccctggg ccaacttttg gcgaaaatga
     2581 gacgttgatc ggcacgtaag aggttccaac tttcaccata atgaaataag atcactaccg
     2641 ggcgtatttt ttgagttatc gagattttca ggagctaagg aagctaaaat ggagaaaaaa
     2701 atcactggat ataccaccgt tgatatatcc caatggcatc gtaaagaaca ttttgaggca
     2761 tttcagtcag ttgctcaatg tacctataac cagaccgttc agctggatat tacggccttt
     2821 ttaaagaccg taaagaaaaa taagcacaag ttttatccgg cctttattca cattcttgcc
     2881 cgcctgatga atgctcatcc ggaatttcgt atggcaatga aagacggtga gctggtgata
     2941 tgggatagtg ttcacccttg ttacaccgtt ttccatgagc aaactgaaac gttttcatcg
     3001 ctctggagtg aataccacga cgatttccgg cagtttctac acatatattc gcaagatgtg
     3061 gcgtgttacg gtgaaaacct ggcctatttc cctaaagggt ttattgagaa tatgtttttc
     3121 gtctcagcca atccctgggt gagtttcacc agttttgatt taaacgtggc caatatggac
     3181 aacttcttcg cccccgtttt cacgatgggc aaatattata cgcaaggcga caaggtgctg
     3241 atgccgctgg cgattcaggt tcatcatgcc gtttgtgatg gcttccatgt cggcagaatg
     3301 cttaatgaat tacaacagta ctgcgatgag tggcagggcg gggcgtaatt tttttaaggc
     3361 agttattggt gcccttaaac gcctggttgc tacgcctgaa taagtgataa taagcggatg
     3421 aatggcagaa attcgatgat aagctgtcaa acatgagaat tggtctcgac gagtatgctt
     3481 ggattctcac caataaaaaa cgcccggcgg caaccgagcg ttctgaacaa atccagatgg
     3541 agttctgagg tcattactgg atctatcaac aggagtccaa gcgacgtctg tgcaagtact
     3601 actgttctgc agtcacttga attcgatacc cagctgttat ttgtatagtt catccatgcc
     3661 atgtgtaatc ccagcagcgg ttacaaactc aagaaggacc atgtggtctc tcttttcgtt
     3721 gggatctttc gaaagggcag attgtgtgga caggtaatgg ttgtctggta aaaggacagg
     3781 gccatcgcca atcggagtat tttgttgata atggtctgct agttgaacgc ttccatcttc
     3841 aatgttgtgt ctaattttga agttaacttt gattccattc ttttgtttgt ctgccatgat
     3901 gtatacattg tgtgagttat agttgtattc caatttgtgt ccaagaatgt ttccatcttc
     3961 tttaaaatca atacctttta actcgattct attaacaagg gtatcacctt caaacttgac
     4021 ttcagcacgt gtcttgtagt tcccgtcatc tttgaaaaat atagttcttt cctgtacgta
     4081 accttcgggc atggcactct tgaaaaagtc atgctgtttc atgtgatctg ggtatctcgc
     4141 aaagcattga acaccataac cgaaagtagt gacaagtgtt ggccacggaa caggtagttt
     4201 tccagtagtg caaataaatt taagggtaag ttttccgtat gttgcatcac cttcaccctc
     4261 tccactgaca gaaaatttgt gcccattaac atcaccatct aattcaacaa gaattgggac
     4321 aactccagtg aaaagttctt ctcctttacg catattatac ctccttaatt tagtgtcatc
     4381 tacaactggc atagtagtct cctgaggttt tattagaatc attatgtttg agcgtagcgt
     4441 tggtaacgct accaggcgcg tgaatgttac cccattttat gtatggacat gtagtgaaaa
     4501 ctacaacctt gtcgttcgct ttggcatcat tttcattata aagacggaat tttgcatcag
     4561 cttctcatca agaaaaacaa ctaaaaaatc aatcacaata cttgtgcatt tctttaaaca
     4621 gtttttaaaa actgactccg ggtatggagc tatgggtatt ttctgtaccc aatgctttta
     4681 acagcaatta atttcatagg atgaaagctc acctagggac cgtctcgaga gaatcaatat
     4741 taatccaacg cgtggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt
     4801 tttatctgtt gtttgtcggt gaacgctctc ctgagtagga caaatccgcc gccctagact
     4861 taggcgttcg gctgcggcga gcggtatcag ctcactcaaa gaagtggggc ccagagagaa
     4921 tattcaggcc agttatgctt tctggcctgt aacaaaggac attaagtaaa gacagataaa
     4981 cgtagactaa aacgtggtcg catcagggtg ctggcttttc aagttcctta agaatggcct
     5041 caattttctc tatacactca gttggaacac gggacctgtc caggttaagc accattttat
     5101 cgcccttata caatactgtc gctccaggag caaactgatg tcgtgagctt aaactagttc
     5161 ttgatgcaga tgacgtttta agcacagaag ttaaaagagt gataacttct tcagcttcaa
     5221 atatcacccc agcttttttc tgctcatgaa ggttagatgc ctgctgctta agtaattcct
     5281 ctttatctgt aaaggctttt tgaagtgcat cacctgaccg ggcagatagt tcaccggggt
     5341 gagaaaaaag agcaacaact gatttaggca atttggcggt gttgatacag cgggtaataa
     5401 tcttacgtga aatattttcc gcatcagcca gcgcagaaat atttccagca aattcattct
     5461 gcaatcggct tgcataacgc tgaccacgtt cataagcact tgttgggcga taatcgttac
     5521 ccaatctgga taatgcagcc atctgctcat catccagctc gccaaccaga acacgataat
     5581 cactttcggt aagtgcagca gctttacgac ggcgactccc atcggcaatt tctatgacac
     5641 cagatactct tcgaccgaac gccggtgtct gttgaccagt cagtagaaaa gaagggatga
     5701 gatcatccag tgcgtcctca gtaagcagct cctggtcacg ttcattacct gaccataccc
     5761 gagaggtctt ctcaacacta tcaccccgga gcacttcaag agtaaacttc acatcccgac
     5821 cacatacagg caaagtaatg gcattaccgc gagccattac tcctacgcgc gcaattaacg
     5881 aatccaccat cggggcagct ggtgtcgata acgaagtatc ttcaaccggt tgagtattga
     5941 gcgtatgttt tggaataaca ggcgcacgct tcattatcta atctcccagc gtggtttaat
     6001 cagacgatcg aaaatttcat tgcagacagg ttcccaaata gaaagagcat ttctccaggc
     6061 accagttgaa gagcgttgat caatggcctg ttcaaaaaca gttctcatcc ggatctgacc
     6121 tttaccaact tcatccgttt cacgtacaac attttttaga accatgcttc cccaggcatc
     6181 ccgaatttgc tcctccatcc acggggactg agagccatta ctattgctgt atttggtaag
     6241 caaaatacgt acatcaggct cgaacccttt aagatcaacg ttcttgagca gatcacgaag
     6301 catatcgaaa aactgcagtg cggaggtgta gtcaaacaac tcagcaggcg tgggaacaat
     6361 cagcacatca gcagcacata cgacattaat cgtgccgata cccaggttag gcgcgctgtc
     6421 aataactatg acatcatagt catgagcaac agtttcaatg gccagtcgga gcatcaggtg
     6481 tggatcggtg ggcagtttac cttcatcaaa tttgcccatt aactcagttt caatacggtg
     6541 cagagccaga caggaaggaa taatgtcaag ccccggccag caagtgggct ttattgcata
     6601 agtgacatcg tccttttccc caagatagaa aggcaggaga gtgtcttctg catgaatatg
     6661 aagatctggt acccatccgt gatacattga ggctgttccc tgggggtcgt taccttccac
     6721 gagcaaaaca cgtagcccct tcagagccag atcctgagca agatgaacag aaactgaggt
     6781 tttgtaaacg ccacctttat gggcagcaac cccgatcacc ggtggaaata cgtcttcagc
     6841 acgtcgcaat cgcgtaccaa acacatcacg catatgatta atttgttcaa ttgtataacc
     6901 aacacgttgc tcaacccgtc ctcgaatttc catatccggg tgcggtagtc gccctgcttt
     6961 ctcggcatct ctgatagcct gagaagaaac cccaactaaa tccgctgctt cacctattct
     7021 ccagcgccgg gttattttcc tcgcttccgg gctgtcatca ttaaactgtg caatggcgat
     7081 agccttcgtc atttcatgac cagcgtttat gcactggtta agtgtttcca tgagtttcat
     7141 tctgaacatc ctttaatcat tgctttgcgt ttttttatta aatcttgcaa tttactgcaa
     7201 agcaacaaca aaatcgcaaa gtcatcaaaa aaccgcaaag ttgtttaaaa taagagcaac
     7261 actacaaaag gagataagaa gagcacatac ctcagtcact tattatcact agcgctcgcc
     7321 gcagccgtgt aaccgagcat agcgagcgaa ctggcgagga agcaaagaag aactgttctg
     7381 tcagatagct cttacgctca gcgcaagaag aaatatccac cgtgggaaaa actccaggta
     7441 gaggtacaca cgcggatagc caattcagag taataaactg tgataatcaa ccctcatcaa
     7501 tgatgacgaa ctaacccccg atatcaagtc acatgacgaa gggaaagaga aggaaatcaa
     7561 ctgtgacaaa ctgccctcaa atttggcttc cttaaaaatt acagttcaaa aagtatgaga
     7621 aaatccatgc aggctgaagg aaacagcaaa actgtgacaa attaccctca gtaggtcaga
     7681 acaaatgtga cgaaccaccc tcaaatctgt gacagataac cctcagacta tcctgtcgtc
     7741 atggaagtga tatcg
//
