LOCUS       sequence-441277-        7643 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To track the natural NsrR promoter's transcriptional dynamics.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7643)
  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 7643)
  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: NsrR-GFP
FEATURES             Location/Qualifiers
     source          1..7643
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             746..1717
                     /codon_start=1
                     /gene="sopB"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopB
                     /translation="MKRAPVIPKHTLNTQPVEDTSLSTPAAPMVDSLIARVGVMARGNA
                     ITLPVCGRDVKFTLEVLRGDSVEKTSRVWSGNERDQELLTEDALDDLIPSFLLTGQQTP
                     AFGRRVSGVIEIADGSRRRKAAALTESDYRVLVGELDDEQMAALSRLGNDYRPTSAYER
                     GQRYASRLQNEFAGNISALADAENISRKIITRCINTAKLPKSVVALFSHPGELSARSGD
                     ALQKAFTDKEELLKQQASNLHEQKKAGVIFEAEEVITLLTSVLKTSSASRTSLSSRHQF
                     APGATVLYKGDKMVLNLDRSRVPTECIEKIEAILKELEKPAP"
     terminator      complement(1875..1961)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     RBS             2241..2249
                     /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             2255..2968
                     /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(2285..2313)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2908..2928
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      3031..3125
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3260..3919)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3834..3853
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(3920..4022)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     rep_origin      4880..5494
                     /label=oriV
                     /note="origin of replication for the bacterial F plasmid"
     rep_origin      4971..5502
                     /label=oriV
                     /note="incP origin of replication"
     rep_origin      5570..5789
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     CDS             5880..6635
                     /codon_start=1
                     /gene="repE"
                     /product="replication initiation protein for the bacterial 
                     F plasmid"
                     /label=repE
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     misc_feature    6638..6888
                     /gene="incC"
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     CDS             join(7214..7643,1..746)
                     /codon_start=1
                     /gene="sopA"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopA
                     /translation="MFRMKLMETLNQCINAGHEMTKAIAIAQFNDDSPEARKITRRWRI
                     GEAADLVGVSSQAIRDAEKAGRLPHPDMEIRGRVEQRVGYTIEQINHMRDVFGTRLRRA
                     EDVFPPVIGVAAHKGGVYKTSVSVHLAQDLALKGLRVLLVEGNDPQGTASMYHGWVPDL
                     HIHAEDTLLPFYLGEKDDVTYAIKPTCWPGLDIIPSCLALHRIETELMGKFDEGKLPTD
                     PHLMLRLAIETVAHDYDVIVIDSAPNLGIGTINVVCAADVLIVPTPAELFDYTSALQFF
                     DMLRDLLKNVDLKGFEPDVRILLTKYSNSNGSQSPWMEEQIRDAWGSMVLKNVVRETDE
                     VGKGQIRMRTVFEQAIDQRSSTGAWRNALSIWEPVCNEIFDRLIKPRWEIR"
ORIGIN
        1 tggaaggtaa cgacccccag ggaacagcct caatgtatca cggatgggta ccagatcttc
       61 atattcatgc agaagacact ctcctgcctt tctatcttgg ggaaaaggac gatgtcactt
      121 atgcaataaa gcccacttgc tggccggggc ttgacattat tccttcctgt ctggctctgc
      181 accgtattga aactgagtta atgggcaaat ttgatgaagg taaactgccc accgatccac
      241 acctgatgct ccgactggcc attgaaactg ttgctcatga ctatgatgtc atagttattg
      301 acagcgcgcc taacctgggt atcggcacga ttaatgtcgt atgtgctgct gatgtgctga
      361 ttgttcccac gcctgctgag ttgtttgact acacctccgc actgcagttt ttcgatatgc
      421 ttcgtgatct gctcaagaac gttgatctta aagggttcga gcctgatgta cgtattttgc
      481 ttaccaaata cagcaatagt aatggctctc agtccccgtg gatggaggag caaattcggg
      541 atgcctgggg aagcatggtt ctaaaaaatg ttgtacgtga aacggatgaa gttggtaaag
      601 gtcagatccg gatgagaact gtttttgaac aggccattga tcaacgctct tcaactggtg
      661 cctggagaaa tgctctttct atttgggaac ctgtctgcaa tgaaattttc gatcgtctga
      721 ttaaaccacg ctgggagatt agataatgaa gcgtgcgcct gttattccaa aacatacgct
      781 caatactcaa ccggttgaag atacttcgtt atcgacacca gctgccccga tggtggattc
      841 gttaattgcg cgcgtaggag taatggctcg cggtaatgcc attactttgc ctgtatgtgg
      901 tcgggatgtg aagtttactc ttgaagtgct ccggggtgat agtgttgaga agacctctcg
      961 ggtatggtca ggtaatgaac gtgaccagga gctgcttact gaggacgcac tggatgatct
     1021 catcccttct tttctactga ctggtcaaca gacaccggcg ttcggtcgaa gagtatctgg
     1081 tgtcatagaa attgccgatg ggagtcgccg tcgtaaagct gctgcactta ccgaaagtga
     1141 ttatcgtgtt ctggttggcg agctggatga tgagcagatg gctgcattat ccagattggg
     1201 taacgattat cgcccaacaa gtgcttatga acgtggtcag cgttatgcaa gccgattgca
     1261 gaatgaattt gctggaaata tttctgcgct ggctgatgcg gaaaatattt cacgtaagat
     1321 tattacccgc tgtatcaaca ccgccaaatt gcctaaatca gttgttgctc ttttttctca
     1381 ccccggtgaa ctatctgccc ggtcaggtga tgcacttcaa aaagccttta cagataaaga
     1441 ggaattactt aagcagcagg catctaacct tcatgagcag aaaaaagctg gggtgatatt
     1501 tgaagctgaa gaagttatca ctcttttaac ttctgtgctt aaaacgtcat ctgcatcaag
     1561 aactagttta agctcacgac atcagtttgc tcctggagcg acagtattgt ataagggcga
     1621 taaaatggtg cttaacctgg acaggtcccg tgttccaact gagtgtatag agaaaattga
     1681 ggccattctt aaggaacttg aaaagccagc accctgatgc gaccacgttt tagtctacgt
     1741 ttatctgtct ttacttaatg tcctttgtta caggccagaa agcataactg gcctgaatat
     1801 tctctctggg ccccacttct ttgagtgagc tgataccgct cgccgcagcc gaacgcctaa
     1861 gtctagggcg gcggatttgt cctactcagg agagcgttca ccgacaaaca acagataaaa
     1921 cgaaaggccc agtctttcga ctgagccttt cgttttattt gatgccacgc gttggattaa
     1981 tattgattct ctcgagacgg tccctaggta attctggtac gcctggcaga tattttgcct
     2041 gccgggcgaa cagtgtgata cattgctgtg tcgggtaagc cattacgcta tccgacacag
     2101 tgttaaatcc tcgctttttt ccttccccga actgaaataa attagcgaca cagcttgtgg
     2161 ctggtttatc atcaatataa atgtattttt tcccgatttc ccttttgagg ttgatatgcc
     2221 agttgtagat gacactaaat taaggaggta taatatgcgt aaaggagaag aacttttcac
     2281 tggagttgtc ccaattcttg ttgaattaga tggtgatgtt aatgggcaca aattttctgt
     2341 cagtggagag ggtgaaggtg atgcaacata cggaaaactt acccttaaat ttatttgcac
     2401 tactggaaaa ctacctgttc cgtggccaac acttgtcact actttcggtt atggtgttca
     2461 atgctttgcg agatacccag atcacatgaa acagcatgac tttttcaaga gtgccatgcc
     2521 cgaaggttac gtacaggaaa gaactatatt tttcaaagat gacgggaact acaagacacg
     2581 tgctgaagtc aagtttgaag gtgataccct tgttaataga atcgagttaa aaggtattga
     2641 ttttaaagaa gatggaaaca ttcttggaca caaattggaa tacaactata actcacacaa
     2701 tgtatacatc atggcagaca aacaaaagaa tggaatcaaa gttaacttca aaattagaca
     2761 caacattgaa gatggaagcg ttcaactagc agaccattat caacaaaata ctccgattgg
     2821 cgatggccct gtccttttac cagacaacca ttacctgtcc acacaatctg ccctttcgaa
     2881 agatcccaac gaaaagagag accacatggt ccttcttgag tttgtaaccg ctgctgggat
     2941 tacacatggc atggatgaac tatacaaata acagctgggt atcgaattca agtgactgca
     3001 gaacagtagt acttgcacag acgtcgcttg gactcctgtt gatagatcca gtaatgacct
     3061 cagaactcca tctggatttg ttcagaacgc tcggttgccg ccgggcgttt tttattggtg
     3121 agaatccaag catactcgtc gagaccaatt ctcatgtttg acagcttatc atcgaatttc
     3181 tgccattcat ccgcttatta tcacttattc aggcgtagca accaggcgtt taagggcacc
     3241 aataactgcc ttaaaaaaat tacgccccgc cctgccactc atcgcagtac tgttgtaatt
     3301 cattaagcat tctgccgaca tggaagccat cacaaacggc atgatgaacc tgaatcgcca
     3361 gcggcatcag caccttgtcg ccttgcgtat aatatttgcc catcgtgaaa acgggggcga
     3421 agaagttgtc catattggcc acgtttaaat caaaactggt gaaactcacc cagggattgg
     3481 ctgagacgaa aaacatattc tcaataaacc ctttagggaa ataggccagg ttttcaccgt
     3541 aacacgccac atcttgcgaa tatatgtgta gaaactgccg gaaatcgtcg tggtattcac
     3601 tccagagcga tgaaaacgtt tcagtttgct catggaaaac ggtgtaacaa gggtgaacac
     3661 tatcccatat caccagctca ccgtctttca ttgccatacg aaattccgga tgagcattca
     3721 tcaggcgggc aagaatgtga ataaaggccg gataaaactt gtgcttattt ttctttacgg
     3781 tctttaaaaa ggccgtaata tccagctgaa cggtctggtt ataggtacat tgagcaactg
     3841 actgaaatgc ctcaaaatgt tctttacgat gccattggga tatatcaacg gtggtatatc
     3901 cagtgatttt tttctccatt ttagcttcct tagctcctga aaatctcgat aactcaaaaa
     3961 atacgcccgg tagtgatctt atttcattat ggtgaaagtt ggaacctctt acgtgccgat
     4021 caacgtctca ttttcgccaa aagttggccc agggcttccc ggtatcaaca gggacaccag
     4081 gatttattta ttctgcgaag tgatcttccg tcacaggtat ttattcgcga taagctcatg
     4141 gagcggcgta accgtcgcac aggaaggaca gagaaagcgc ggatctggga agtgacggac
     4201 agaacggtca ggacctggat tggggaggcg gttgccgccg ctgctgctga cggtgtgacg
     4261 ttctctgttc cggtcacacc acatacgttc cgccattcct atgcgatgca catgctgtat
     4321 gccggtatac cgctgaaagt tctgcaaagc ctgatgggac ataagtccat cagttcaacg
     4381 gaagtctaca cgaaggtttt tgcgctggat gtggctgccc ggcaccgggt gcagtttgcg
     4441 atgccggagt ctgatgcggt tgcgatgctg aaacaattat cctgagaata aatgccttgg
     4501 cctttatatg gaaatgtgga actgagtgga tatgctgttt ttgtctgtta aacagagaag
     4561 ctggctgtta tccactgaga agcgaacgaa acagtcggga aaatctccca ttatcgtaga
     4621 gatccgcatt attaatctca ggagcctgtg tagcgtttat aggaagtagt gttctgtcat
     4681 gatgccagca agcggtaacg aaaacgattt gaatatgcct tcaggaacaa tagaaatctt
     4741 cgtgcggtgt tacgttgaag tggagcggat tatgtcagca atggacagaa caacctaatg
     4801 aacacagaac catgatgtgg tctgtccttt tacagccagt agtgctcgcc gcagtcgagc
     4861 gacagggcga agccctcggc tggttgccct cgccgctggg ctggcggccg tctatggccc
     4921 tgcaaacgcg ccagaaacgc cgtcgaagcc gtgtgcgaga caccgcggcc ggccgccggc
     4981 gttgtggata cctcgcggaa aacttggccc tcactgacag atgaggggcg gacgttgaca
     5041 cttgaggggc cgactcaccc ggcgcggcgt tgacagatga ggggcaggct cgatttcggc
     5101 cggcgacgtg gagctggcca gcctcgcaaa tcggcgaaaa cgcctgattt tacgcgagtt
     5161 tcccacagat gatgtggaca agcctgggga taagtgccct gcggtattga cacttgaggg
     5221 gcgcgactac tgacagatga ggggcgcgat ccttgacact tgaggggcag agtgctgaca
     5281 gatgaggggc gcacctattg acatttgagg ggctgtccac aggcagaaaa tccagcattt
     5341 gcaagggttt ccgcccgttt ttcggccacc gctaacctgt cttttaacct gcttttaaac
     5401 caatatttat aaaccttgtt tttaaccagg gctgcgccct gtgcgcgtga ccgcgcacgc
     5461 cgaagggggg tgccccccct tctcgaaccc tcccggtcga gtgagcgagg aagcaccagg
     5521 gaacagcact tatatattct gcttacacac gatgcctgaa aaaacttccc ttggggttat
     5581 ccacttatcc acggggatat ttttataatt atttttttta tagtttttag atcttctttt
     5641 ttagagcgcc ttgtaggcct ttatccatgc tggttctaga gaaggtgttg tgacaaattg
     5701 ccctttcagt gtgacaaatc accctcaaat gacagtcctg tctgtgacaa attgccctta
     5761 accctgtgac aaattgccct cagaagaagc tgttttttca caaagttatc cctgcttatt
     5821 gactcttttt tatttagtgt gacaatctaa aaacttgtca cacttcacat ggatctgtca
     5881 tggcggaaac agcggttatc aatcacaaga aacgtaaaaa tagcccgcga atcgtccagt
     5941 caaacgacct cactgaggcg gcatatagtc tctcccggga tcaaaaacgt atgctgtatc
     6001 tgttcgttga ccagatcaga aaatctgatg gcaccctaca ggaacatgac ggtatctgcg
     6061 agatccatgt tgctaaatat gctgaaatat tcggattgac ctctgcggaa gccagtaagg
     6121 atatacggca ggcattgaag agtttcgcgg ggaaggaagt ggttttttat cgccctgaag
     6181 aggatgccgg cgatgaaaaa ggctatgaat cttttccttg gtttatcaaa cgtgcgcaca
     6241 gtccatccag agggctttac agtgtacata tcaacccata tctcattccc ttctttatcg
     6301 ggttacagaa ccggtttacg cagtttcggc ttagtgaaac aaaagaaatc accaatccgt
     6361 atgccatgcg tttatacgaa tccctgtgtc agtatcgtaa gccggatggc tcaggcatcg
     6421 tctctctgaa aatcgactgg atcatagagc gttaccagct gcctcaaagt taccagcgta
     6481 tgcctgactt ccgccgccgc ttcctgcagg tctgtgttaa tgagatcaac agcagaactc
     6541 caatgcgcct ctcatacatt gagaaaaaga aaggccgcca gacgactcat atcgtatttt
     6601 ccttccgcga tatcacttcc atgacgacag gatagtctga gggttatctg tcacagattt
     6661 gagggtggtt cgtcacattt gttctgacct actgagggta atttgtcaca gttttgctgt
     6721 ttccttcagc ctgcatggat tttctcatac tttttgaact gtaattttta aggaagccaa
     6781 atttgagggc agtttgtcac agttgatttc cttctctttc ccttcgtcat gtgacttgat
     6841 atcgggggtt agttcgtcat cattgatgag ggttgattat cacagtttat tactctgaat
     6901 tggctatccg cgtgtgtacc tctacctgga gtttttccca cggtggatat ttcttcttgc
     6961 gctgagcgta agagctatct gacagaacag ttcttctttg cttcctcgcc agttcgctcg
     7021 ctatgctcgg ttacacggct gcggcgagcg ctagtgataa taagtgactg aggtatgtgc
     7081 tcttcttatc tccttttgta gtgttgctct tattttaaac aactttgcgg ttttttgatg
     7141 actttgcgat tttgttgttg ctttgcagta aattgcaaga tttaataaaa aaacgcaaag
     7201 caatgattaa aggatgttca gaatgaaact catggaaaca cttaaccagt gcataaacgc
     7261 tggtcatgaa atgacgaagg ctatcgccat tgcacagttt aatgatgaca gcccggaagc
     7321 gaggaaaata acccggcgct ggagaatagg tgaagcagcg gatttagttg gggtttcttc
     7381 tcaggctatc agagatgccg agaaagcagg gcgactaccg cacccggata tggaaattcg
     7441 aggacgggtt gagcaacgtg ttggttatac aattgaacaa attaatcata tgcgtgatgt
     7501 gtttggtacg cgattgcgac gtgctgaaga cgtatttcca ccggtgatcg gggttgctgc
     7561 ccataaaggt ggcgtttaca aaacctcagt ttctgttcat cttgctcagg atctggctct
     7621 gaaggggcta cgtgttttgc tcg
//
