LOCUS       sequence-439541-        7647 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To track the natural MarA promoter's transcriptional dynamics.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7647)
  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 7647)
  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: MarA-GFP
FEATURES             Location/Qualifiers
     source          1..7647
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    193..443
                     /gene="incC"
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     CDS             complement(446..1201)
                     /codon_start=1
                     /gene="repE"
                     /product="replication initiation protein for the bacterial 
                     F plasmid"
                     /label=repE
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     rep_origin      1292..1511
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     rep_origin      1579..2110
                     /label=oriV
                     /note="incP origin of replication"
     rep_origin      1587..2201
                     /label=oriV
                     /note="origin of replication for the bacterial F plasmid"
     promoter        3059..3161
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             3162..3821
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(3228..3247)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      complement(3956..4050)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(4113..4826)
                     /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(4153..4173)
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     4768..4796
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     RBS             4832..4840
                     /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      5124..5210
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(5368..6339)
                     /codon_start=1
                     /gene="sopB"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopB
                     /translation="MKRAPVIPKHTLNTQPVEDTSLSTPAAPMVDSLIARVGVMARGNA
                     ITLPVCGRDVKFTLEVLRGDSVEKTSRVWSGNERDQELLTEDALDDLIPSFLLTGQQTP
                     AFGRRVSGVIEIADGSRRRKAAALTESDYRVLVGELDDEQMAALSRLGNDYRPTSAYER
                     GQRYASRLQNEFAGNISALADAENISRKIITRCINTAKLPKSVVALFSHPGELSARSGD
                     ALQKAFTDKEELLKQQASNLHEQKKAGVIFEAEEVITLLTSVLKTSSASRTSLSSRHQF
                     APGATVLYKGDKMVLNLDRSRVPTECIEKIEAILKELEKPAP"
     CDS             complement(6339..7514)
                     /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 gcacatacct cagtcactta ttatcactag cgctcgccgc agccgtgtaa ccgagcatag
       61 cgagcgaact ggcgaggaag caaagaagaa ctgttctgtc agatagctct tacgctcagc
      121 gcaagaagaa atatccaccg tgggaaaaac tccaggtaga ggtacacacg cggatagcca
      181 attcagagta ataaactgtg ataatcaacc ctcatcaatg atgacgaact aacccccgat
      241 atcaagtcac atgacgaagg gaaagagaag gaaatcaact gtgacaaact gccctcaaat
      301 ttggcttcct taaaaattac agttcaaaaa gtatgagaaa atccatgcag gctgaaggaa
      361 acagcaaaac tgtgacaaat taccctcagt aggtcagaac aaatgtgacg aaccaccctc
      421 aaatctgtga cagataaccc tcagactatc ctgtcgtcat ggaagtgata tcgcggaagg
      481 aaaatacgat atgagtcgtc tggcggcctt tctttttctc aatgtatgag aggcgcattg
      541 gagttctgct gttgatctca ttaacacaga cctgcaggaa gcggcggcgg aagtcaggca
      601 tacgctggta actttgaggc agctggtaac gctctatgat ccagtcgatt ttcagagaga
      661 cgatgcctga gccatccggc ttacgatact gacacaggga ttcgtataaa cgcatggcat
      721 acggattggt gatttctttt gtttcactaa gccgaaactg cgtaaaccgg ttctgtaacc
      781 cgataaagaa gggaatgaga tatgggttga tatgtacact gtaaagccct ctggatggac
      841 tgtgcgcacg tttgataaac caaggaaaag attcatagcc tttttcatcg ccggcatcct
      901 cttcagggcg ataaaaaacc acttccttcc ccgcgaaact cttcaatgcc tgccgtatat
      961 ccttactggc ttccgcagag gtcaatccga atatttcagc atatttagca acatggatct
     1021 cgcagatacc gtcatgttcc tgtagggtgc catcagattt tctgatctgg tcaacgaaca
     1081 gatacagcat acgtttttga tcccgggaga gactatatgc cgcctcagtg aggtcgtttg
     1141 actggacgat tcgcgggcta tttttacgtt tcttgtgatt gataaccgct gtttccgcca
     1201 tgacagatcc atgtgaagtg tgacaagttt ttagattgtc acactaaata aaaaagagtc
     1261 aataagcagg gataactttg tgaaaaaaca gcttcttctg agggcaattt gtcacagggt
     1321 taagggcaat ttgtcacaga caggactgtc atttgagggt gatttgtcac actgaaaggg
     1381 caatttgtca caacaccttc tctagaacca gcatggataa aggcctacaa ggcgctctaa
     1441 aaaagaagat ctaaaaacta taaaaaaaat aattataaaa atatccccgt ggataagtgg
     1501 ataaccccaa gggaagtttt ttcaggcatc gtgtgtaagc agaatatata agtgctgttc
     1561 cctggtgctt cctcgctcac tcgaccggga gggttcgaga agggggggca ccccccttcg
     1621 gcgtgcgcgg tcacgcgcac agggcgcagc cctggttaaa aacaaggttt ataaatattg
     1681 gtttaaaagc aggttaaaag acaggttagc ggtggccgaa aaacgggcgg aaacccttgc
     1741 aaatgctgga ttttctgcct gtggacagcc cctcaaatgt caataggtgc gcccctcatc
     1801 tgtcagcact ctgcccctca agtgtcaagg atcgcgcccc tcatctgtca gtagtcgcgc
     1861 ccctcaagtg tcaataccgc agggcactta tccccaggct tgtccacatc atctgtggga
     1921 aactcgcgta aaatcaggcg ttttcgccga tttgcgaggc tggccagctc cacgtcgccg
     1981 gccgaaatcg agcctgcccc tcatctgtca acgccgcgcc gggtgagtcg gcccctcaag
     2041 tgtcaacgtc cgcccctcat ctgtcagtga gggccaagtt ttccgcgagg tatccacaac
     2101 gccggcggcc ggccgcggtg tctcgcacac ggcttcgacg gcgtttctgg cgcgtttgca
     2161 gggccataga cggccgccag cccagcggcg agggcaacca gccgagggct tcgccctgtc
     2221 gctcgactgc ggcgagcact actggctgta aaaggacaga ccacatcatg gttctgtgtt
     2281 cattaggttg ttctgtccat tgctgacata atccgctcca cttcaacgta acaccgcacg
     2341 aagatttcta ttgttcctga aggcatattc aaatcgtttt cgttaccgct tgctggcatc
     2401 atgacagaac actacttcct ataaacgcta cacaggctcc tgagattaat aatgcggatc
     2461 tctacgataa tgggagattt tcccgactgt ttcgttcgct tctcagtgga taacagccag
     2521 cttctctgtt taacagacaa aaacagcata tccactcagt tccacatttc catataaagg
     2581 ccaaggcatt tattctcagg ataattgttt cagcatcgca accgcatcag actccggcat
     2641 cgcaaactgc acccggtgcc gggcagccac atccagcgca aaaaccttcg tgtagacttc
     2701 cgttgaactg atggacttat gtcccatcag gctttgcaga actttcagcg gtataccggc
     2761 atacagcatg tgcatcgcat aggaatggcg gaacgtatgt ggtgtgaccg gaacagagaa
     2821 cgtcacaccg tcagcagcag cggcggcaac cgcctcccca atccaggtcc tgaccgttct
     2881 gtccgtcact tcccagatcc gcgctttctc tgtccttcct gtgcgacggt tacgccgctc
     2941 catgagctta tcgcgaataa atacctgtga cggaagatca cttcgcagaa taaataaatc
     3001 ctggtgtccc tgttgatacc gggaagccct gggccaactt ttggcgaaaa tgagacgttg
     3061 atcggcacgt aagaggttcc aactttcacc ataatgaaat aagatcacta ccgggcgtat
     3121 tttttgagtt atcgagattt tcaggagcta aggaagctaa aatggagaaa aaaatcactg
     3181 gatataccac cgttgatata tcccaatggc atcgtaaaga acattttgag gcatttcagt
     3241 cagttgctca atgtacctat aaccagaccg ttcagctgga tattacggcc tttttaaaga
     3301 ccgtaaagaa aaataagcac aagttttatc cggcctttat tcacattctt gcccgcctga
     3361 tgaatgctca tccggaattt cgtatggcaa tgaaagacgg tgagctggtg atatgggata
     3421 gtgttcaccc ttgttacacc gttttccatg agcaaactga aacgttttca tcgctctgga
     3481 gtgaatacca cgacgatttc cggcagtttc tacacatata ttcgcaagat gtggcgtgtt
     3541 acggtgaaaa cctggcctat ttccctaaag ggtttattga gaatatgttt ttcgtctcag
     3601 ccaatccctg ggtgagtttc accagttttg atttaaacgt ggccaatatg gacaacttct
     3661 tcgcccccgt tttcacgatg ggcaaatatt atacgcaagg cgacaaggtg ctgatgccgc
     3721 tggcgattca ggttcatcat gccgtttgtg atggcttcca tgtcggcaga atgcttaatg
     3781 aattacaaca gtactgcgat gagtggcagg gcggggcgta atttttttaa ggcagttatt
     3841 ggtgccctta aacgcctggt tgctacgcct gaataagtga taataagcgg atgaatggca
     3901 gaaattcgat gataagctgt caaacatgag aattggtctc gacgagtatg cttggattct
     3961 caccaataaa aaacgcccgg cggcaaccga gcgttctgaa caaatccaga tggagttctg
     4021 aggtcattac tggatctatc aacaggagtc caagcgacgt ctgtgcaagt actactgttc
     4081 tgcagtcact tgaattcgat acccagctgt tatttgtata gttcatccat gccatgtgta
     4141 atcccagcag cggttacaaa ctcaagaagg accatgtggt ctctcttttc gttgggatct
     4201 ttcgaaaggg cagattgtgt ggacaggtaa tggttgtctg gtaaaaggac agggccatcg
     4261 ccaatcggag tattttgttg ataatggtct gctagttgaa cgcttccatc ttcaatgttg
     4321 tgtctaattt tgaagttaac tttgattcca ttcttttgtt tgtctgccat gatgtataca
     4381 ttgtgtgagt tatagttgta ttccaatttg tgtccaagaa tgtttccatc ttctttaaaa
     4441 tcaatacctt ttaactcgat tctattaaca agggtatcac cttcaaactt gacttcagca
     4501 cgtgtcttgt agttcccgtc atctttgaaa aatatagttc tttcctgtac gtaaccttcg
     4561 ggcatggcac tcttgaaaaa gtcatgctgt ttcatgtgat ctgggtatct cgcaaagcat
     4621 tgaacaccat aaccgaaagt agtgacaagt gttggccacg gaacaggtag ttttccagta
     4681 gtgcaaataa atttaagggt aagttttccg tatgttgcat caccttcacc ctctccactg
     4741 acagaaaatt tgtgcccatt aacatcacca tctaattcaa caagaattgg gacaactcca
     4801 gtgaaaagtt cttctccttt acgcatatta tacctcctta atttagtgtc atctacaact
     4861 ggcatattag ttgccctggc aagtaattag ttgcagggga taatattgcc caggcaagta
     4921 taagtcaaat gaatgaattg accgatgcca cgttttgcta aatcggttca agttttgttc
     4981 aatgcgatgc aaggctagct aaagagtgac actttatcct tgtttacccc ttatactttt
     5041 cgctgataac ccagatacac aggataacaa ccaccaccta gggaccgtct cgagagaatc
     5101 aatattaatc caacgcgtgg catcaaataa aacgaaaggc tcagtcgaaa gactgggcct
     5161 ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag taggacaaat ccgccgccct
     5221 agacttaggc gttcggctgc ggcgagcggt atcagctcac tcaaagaagt ggggcccaga
     5281 gagaatattc aggccagtta tgctttctgg cctgtaacaa aggacattaa gtaaagacag
     5341 ataaacgtag actaaaacgt ggtcgcatca gggtgctggc ttttcaagtt ccttaagaat
     5401 ggcctcaatt ttctctatac actcagttgg aacacgggac ctgtccaggt taagcaccat
     5461 tttatcgccc ttatacaata ctgtcgctcc aggagcaaac tgatgtcgtg agcttaaact
     5521 agttcttgat gcagatgacg ttttaagcac agaagttaaa agagtgataa cttcttcagc
     5581 ttcaaatatc accccagctt ttttctgctc atgaaggtta gatgcctgct gcttaagtaa
     5641 ttcctcttta tctgtaaagg ctttttgaag tgcatcacct gaccgggcag atagttcacc
     5701 ggggtgagaa aaaagagcaa caactgattt aggcaatttg gcggtgttga tacagcgggt
     5761 aataatctta cgtgaaatat tttccgcatc agccagcgca gaaatatttc cagcaaattc
     5821 attctgcaat cggcttgcat aacgctgacc acgttcataa gcacttgttg ggcgataatc
     5881 gttacccaat ctggataatg cagccatctg ctcatcatcc agctcgccaa ccagaacacg
     5941 ataatcactt tcggtaagtg cagcagcttt acgacggcga ctcccatcgg caatttctat
     6001 gacaccagat actcttcgac cgaacgccgg tgtctgttga ccagtcagta gaaaagaagg
     6061 gatgagatca tccagtgcgt cctcagtaag cagctcctgg tcacgttcat tacctgacca
     6121 tacccgagag gtcttctcaa cactatcacc ccggagcact tcaagagtaa acttcacatc
     6181 ccgaccacat acaggcaaag taatggcatt accgcgagcc attactccta cgcgcgcaat
     6241 taacgaatcc accatcgggg cagctggtgt cgataacgaa gtatcttcaa ccggttgagt
     6301 attgagcgta tgttttggaa taacaggcgc acgcttcatt atctaatctc ccagcgtggt
     6361 ttaatcagac gatcgaaaat ttcattgcag acaggttccc aaatagaaag agcatttctc
     6421 caggcaccag ttgaagagcg ttgatcaatg gcctgttcaa aaacagttct catccggatc
     6481 tgacctttac caacttcatc cgtttcacgt acaacatttt ttagaaccat gcttccccag
     6541 gcatcccgaa tttgctcctc catccacggg gactgagagc cattactatt gctgtatttg
     6601 gtaagcaaaa tacgtacatc aggctcgaac cctttaagat caacgttctt gagcagatca
     6661 cgaagcatat cgaaaaactg cagtgcggag gtgtagtcaa acaactcagc aggcgtggga
     6721 acaatcagca catcagcagc acatacgaca ttaatcgtgc cgatacccag gttaggcgcg
     6781 ctgtcaataa ctatgacatc atagtcatga gcaacagttt caatggccag tcggagcatc
     6841 aggtgtggat cggtgggcag tttaccttca tcaaatttgc ccattaactc agtttcaata
     6901 cggtgcagag ccagacagga aggaataatg tcaagccccg gccagcaagt gggctttatt
     6961 gcataagtga catcgtcctt ttccccaaga tagaaaggca ggagagtgtc ttctgcatga
     7021 atatgaagat ctggtaccca tccgtgatac attgaggctg ttccctgggg gtcgttacct
     7081 tccacgagca aaacacgtag ccccttcaga gccagatcct gagcaagatg aacagaaact
     7141 gaggttttgt aaacgccacc tttatgggca gcaaccccga tcaccggtgg aaatacgtct
     7201 tcagcacgtc gcaatcgcgt accaaacaca tcacgcatat gattaatttg ttcaattgta
     7261 taaccaacac gttgctcaac ccgtcctcga atttccatat ccgggtgcgg tagtcgccct
     7321 gctttctcgg catctctgat agcctgagaa gaaaccccaa ctaaatccgc tgcttcacct
     7381 attctccagc gccgggttat tttcctcgct tccgggctgt catcattaaa ctgtgcaatg
     7441 gcgatagcct tcgtcatttc atgaccagcg tttatgcact ggttaagtgt ttccatgagt
     7501 ttcattctga acatccttta atcattgctt tgcgtttttt tattaaatct tgcaatttac
     7561 tgcaaagcaa caacaaaatc gcaaagtcat caaaaaaccg caaagttgtt taaaataaga
     7621 gcaacactac aaaaggagat aagaaga
//
