LOCUS       sequence-436626-        7604 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To track the natural PhoP promoter's transcriptional dynamics.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7604)
  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 7604)
  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: PhoP-GFP
FEATURES             Location/Qualifiers
     source          1..7604
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             29..1204
                     /codon_start=1
                     /gene="sopA"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopA
                     /translation="MFRMKLMETLNQCINAGHEMTKAIAIAQFNDDSPEARKITRRWRI
                     GEAADLVGVSSQAIRDAEKAGRLPHPDMEIRGRVEQRVGYTIEQINHMRDVFGTRLRRA
                     EDVFPPVIGVAAHKGGVYKTSVSVHLAQDLALKGLRVLLVEGNDPQGTASMYHGWVPDL
                     HIHAEDTLLPFYLGEKDDVTYAIKPTCWPGLDIIPSCLALHRIETELMGKFDEGKLPTD
                     PHLMLRLAIETVAHDYDVIVIDSAPNLGIGTINVVCAADVLIVPTPAELFDYTSALQFF
                     DMLRDLLKNVDLKGFEPDVRILLTKYSNSNGSQSPWMEEQIRDAWGSMVLKNVVRETDE
                     VGKGQIRMRTVFEQAIDQRSSTGAWRNALSIWEPVCNEIFDRLIKPRWEIR"
     CDS             1204..2175
                     /codon_start=1
                     /gene="sopB"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopB
                     /translation="MKRAPVIPKHTLNTQPVEDTSLSTPAAPMVDSLIARVGVMARGNA
                     ITLPVCGRDVKFTLEVLRGDSVEKTSRVWSGNERDQELLTEDALDDLIPSFLLTGQQTP
                     AFGRRVSGVIEIADGSRRRKAAALTESDYRVLVGELDDEQMAALSRLGNDYRPTSAYER
                     GQRYASRLQNEFAGNISALADAENISRKIITRCINTAKLPKSVVALFSHPGELSARSGD
                     ALQKAFTDKEELLKQQASNLHEQKKAGVIFEAEEVITLLTSVLKTSSASRTSLSSRHQF
                     APGATVLYKGDKMVLNLDRSRVPTECIEKIEAILKELEKPAP"
     terminator      complement(2333..2419)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     RBS             2660..2668
                     /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             2674..3387
                     /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(2704..2732)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     3327..3347
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      3450..3544
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3679..4338)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4253..4272
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4339..4441)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     rep_origin      5299..5913
                     /label=oriV
                     /note="origin of replication for the bacterial F plasmid"
     rep_origin      5390..5921
                     /label=oriV
                     /note="incP origin of replication"
     rep_origin      5989..6208
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     CDS             6299..7054
                     /codon_start=1
                     /gene="repE"
                     /product="replication initiation protein for the bacterial 
                     F plasmid"
                     /label=repE
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     misc_feature    7057..7307
                     /gene="incC"
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
ORIGIN
        1 taaaaaaacg caaagcaatg attaaaggat gttcagaatg aaactcatgg aaacacttaa
       61 ccagtgcata aacgctggtc atgaaatgac gaaggctatc gccattgcac agtttaatga
      121 tgacagcccg gaagcgagga aaataacccg gcgctggaga ataggtgaag cagcggattt
      181 agttggggtt tcttctcagg ctatcagaga tgccgagaaa gcagggcgac taccgcaccc
      241 ggatatggaa attcgaggac gggttgagca acgtgttggt tatacaattg aacaaattaa
      301 tcatatgcgt gatgtgtttg gtacgcgatt gcgacgtgct gaagacgtat ttccaccggt
      361 gatcggggtt gctgcccata aaggtggcgt ttacaaaacc tcagtttctg ttcatcttgc
      421 tcaggatctg gctctgaagg ggctacgtgt tttgctcgtg gaaggtaacg acccccaggg
      481 aacagcctca atgtatcacg gatgggtacc agatcttcat attcatgcag aagacactct
      541 cctgcctttc tatcttgggg aaaaggacga tgtcacttat gcaataaagc ccacttgctg
      601 gccggggctt gacattattc cttcctgtct ggctctgcac cgtattgaaa ctgagttaat
      661 gggcaaattt gatgaaggta aactgcccac cgatccacac ctgatgctcc gactggccat
      721 tgaaactgtt gctcatgact atgatgtcat agttattgac agcgcgccta acctgggtat
      781 cggcacgatt aatgtcgtat gtgctgctga tgtgctgatt gttcccacgc ctgctgagtt
      841 gtttgactac acctccgcac tgcagttttt cgatatgctt cgtgatctgc tcaagaacgt
      901 tgatcttaaa gggttcgagc ctgatgtacg tattttgctt accaaataca gcaatagtaa
      961 tggctctcag tccccgtgga tggaggagca aattcgggat gcctggggaa gcatggttct
     1021 aaaaaatgtt gtacgtgaaa cggatgaagt tggtaaaggt cagatccgga tgagaactgt
     1081 ttttgaacag gccattgatc aacgctcttc aactggtgcc tggagaaatg ctctttctat
     1141 ttgggaacct gtctgcaatg aaattttcga tcgtctgatt aaaccacgct gggagattag
     1201 ataatgaagc gtgcgcctgt tattccaaaa catacgctca atactcaacc ggttgaagat
     1261 acttcgttat cgacaccagc tgccccgatg gtggattcgt taattgcgcg cgtaggagta
     1321 atggctcgcg gtaatgccat tactttgcct gtatgtggtc gggatgtgaa gtttactctt
     1381 gaagtgctcc ggggtgatag tgttgagaag acctctcggg tatggtcagg taatgaacgt
     1441 gaccaggagc tgcttactga ggacgcactg gatgatctca tcccttcttt tctactgact
     1501 ggtcaacaga caccggcgtt cggtcgaaga gtatctggtg tcatagaaat tgccgatggg
     1561 agtcgccgtc gtaaagctgc tgcacttacc gaaagtgatt atcgtgttct ggttggcgag
     1621 ctggatgatg agcagatggc tgcattatcc agattgggta acgattatcg cccaacaagt
     1681 gcttatgaac gtggtcagcg ttatgcaagc cgattgcaga atgaatttgc tggaaatatt
     1741 tctgcgctgg ctgatgcgga aaatatttca cgtaagatta ttacccgctg tatcaacacc
     1801 gccaaattgc ctaaatcagt tgttgctctt ttttctcacc ccggtgaact atctgcccgg
     1861 tcaggtgatg cacttcaaaa agcctttaca gataaagagg aattacttaa gcagcaggca
     1921 tctaaccttc atgagcagaa aaaagctggg gtgatatttg aagctgaaga agttatcact
     1981 cttttaactt ctgtgcttaa aacgtcatct gcatcaagaa ctagtttaag ctcacgacat
     2041 cagtttgctc ctggagcgac agtattgtat aagggcgata aaatggtgct taacctggac
     2101 aggtcccgtg ttccaactga gtgtatagag aaaattgagg ccattcttaa ggaacttgaa
     2161 aagccagcac cctgatgcga ccacgtttta gtctacgttt atctgtcttt acttaatgtc
     2221 ctttgttaca ggccagaaag cataactggc ctgaatattc tctctgggcc ccacttcttt
     2281 gagtgagctg ataccgctcg ccgcagccga acgcctaagt ctagggcggc ggatttgtcc
     2341 tactcaggag agcgttcacc gacaaacaac agataaaacg aaaggcccag tctttcgact
     2401 gagcctttcg ttttatttga tgccacgcgt tggattaata ttgattctct cgagacggtc
     2461 cctaggacct cgtatcagtg ccggatggcg atgctgtccg gcctgcttat taagattatc
     2521 cgctttttat tttttcactt tacctcccct ccccgctggt ttatttaatg tttaccccca
     2581 taaccacata atcgcgttac actattttaa taattaagac agggagaaat aaaaatgcca
     2641 gttgtagatg acactaaatt aaggaggtat aatatgcgta aaggagaaga acttttcact
     2701 ggagttgtcc caattcttgt tgaattagat ggtgatgtta atgggcacaa attttctgtc
     2761 agtggagagg gtgaaggtga tgcaacatac ggaaaactta cccttaaatt tatttgcact
     2821 actggaaaac tacctgttcc gtggccaaca cttgtcacta ctttcggtta tggtgttcaa
     2881 tgctttgcga gatacccaga tcacatgaaa cagcatgact ttttcaagag tgccatgccc
     2941 gaaggttacg tacaggaaag aactatattt ttcaaagatg acgggaacta caagacacgt
     3001 gctgaagtca agtttgaagg tgataccctt gttaatagaa tcgagttaaa aggtattgat
     3061 tttaaagaag atggaaacat tcttggacac aaattggaat acaactataa ctcacacaat
     3121 gtatacatca tggcagacaa acaaaagaat ggaatcaaag ttaacttcaa aattagacac
     3181 aacattgaag atggaagcgt tcaactagca gaccattatc aacaaaatac tccgattggc
     3241 gatggccctg tccttttacc agacaaccat tacctgtcca cacaatctgc cctttcgaaa
     3301 gatcccaacg aaaagagaga ccacatggtc cttcttgagt ttgtaaccgc tgctgggatt
     3361 acacatggca tggatgaact atacaaataa cagctgggta tcgaattcaa gtgactgcag
     3421 aacagtagta cttgcacaga cgtcgcttgg actcctgttg atagatccag taatgacctc
     3481 agaactccat ctggatttgt tcagaacgct cggttgccgc cgggcgtttt ttattggtga
     3541 gaatccaagc atactcgtcg agaccaattc tcatgtttga cagcttatca tcgaatttct
     3601 gccattcatc cgcttattat cacttattca ggcgtagcaa ccaggcgttt aagggcacca
     3661 ataactgcct taaaaaaatt acgccccgcc ctgccactca tcgcagtact gttgtaattc
     3721 attaagcatt ctgccgacat ggaagccatc acaaacggca tgatgaacct gaatcgccag
     3781 cggcatcagc accttgtcgc cttgcgtata atatttgccc atcgtgaaaa cgggggcgaa
     3841 gaagttgtcc atattggcca cgtttaaatc aaaactggtg aaactcaccc agggattggc
     3901 tgagacgaaa aacatattct caataaaccc tttagggaaa taggccaggt tttcaccgta
     3961 acacgccaca tcttgcgaat atatgtgtag aaactgccgg aaatcgtcgt ggtattcact
     4021 ccagagcgat gaaaacgttt cagtttgctc atggaaaacg gtgtaacaag ggtgaacact
     4081 atcccatatc accagctcac cgtctttcat tgccatacga aattccggat gagcattcat
     4141 caggcgggca agaatgtgaa taaaggccgg ataaaacttg tgcttatttt tctttacggt
     4201 ctttaaaaag gccgtaatat ccagctgaac ggtctggtta taggtacatt gagcaactga
     4261 ctgaaatgcc tcaaaatgtt ctttacgatg ccattgggat atatcaacgg tggtatatcc
     4321 agtgattttt ttctccattt tagcttcctt agctcctgaa aatctcgata actcaaaaaa
     4381 tacgcccggt agtgatctta tttcattatg gtgaaagttg gaacctctta cgtgccgatc
     4441 aacgtctcat tttcgccaaa agttggccca gggcttcccg gtatcaacag ggacaccagg
     4501 atttatttat tctgcgaagt gatcttccgt cacaggtatt tattcgcgat aagctcatgg
     4561 agcggcgtaa ccgtcgcaca ggaaggacag agaaagcgcg gatctgggaa gtgacggaca
     4621 gaacggtcag gacctggatt ggggaggcgg ttgccgccgc tgctgctgac ggtgtgacgt
     4681 tctctgttcc ggtcacacca catacgttcc gccattccta tgcgatgcac atgctgtatg
     4741 ccggtatacc gctgaaagtt ctgcaaagcc tgatgggaca taagtccatc agttcaacgg
     4801 aagtctacac gaaggttttt gcgctggatg tggctgcccg gcaccgggtg cagtttgcga
     4861 tgccggagtc tgatgcggtt gcgatgctga aacaattatc ctgagaataa atgccttggc
     4921 ctttatatgg aaatgtggaa ctgagtggat atgctgtttt tgtctgttaa acagagaagc
     4981 tggctgttat ccactgagaa gcgaacgaaa cagtcgggaa aatctcccat tatcgtagag
     5041 atccgcatta ttaatctcag gagcctgtgt agcgtttata ggaagtagtg ttctgtcatg
     5101 atgccagcaa gcggtaacga aaacgatttg aatatgcctt caggaacaat agaaatcttc
     5161 gtgcggtgtt acgttgaagt ggagcggatt atgtcagcaa tggacagaac aacctaatga
     5221 acacagaacc atgatgtggt ctgtcctttt acagccagta gtgctcgccg cagtcgagcg
     5281 acagggcgaa gccctcggct ggttgccctc gccgctgggc tggcggccgt ctatggccct
     5341 gcaaacgcgc cagaaacgcc gtcgaagccg tgtgcgagac accgcggccg gccgccggcg
     5401 ttgtggatac ctcgcggaaa acttggccct cactgacaga tgaggggcgg acgttgacac
     5461 ttgaggggcc gactcacccg gcgcggcgtt gacagatgag gggcaggctc gatttcggcc
     5521 ggcgacgtgg agctggccag cctcgcaaat cggcgaaaac gcctgatttt acgcgagttt
     5581 cccacagatg atgtggacaa gcctggggat aagtgccctg cggtattgac acttgagggg
     5641 cgcgactact gacagatgag gggcgcgatc cttgacactt gaggggcaga gtgctgacag
     5701 atgaggggcg cacctattga catttgaggg gctgtccaca ggcagaaaat ccagcatttg
     5761 caagggtttc cgcccgtttt tcggccaccg ctaacctgtc ttttaacctg cttttaaacc
     5821 aatatttata aaccttgttt ttaaccaggg ctgcgccctg tgcgcgtgac cgcgcacgcc
     5881 gaaggggggt gccccccctt ctcgaaccct cccggtcgag tgagcgagga agcaccaggg
     5941 aacagcactt atatattctg cttacacacg atgcctgaaa aaacttccct tggggttatc
     6001 cacttatcca cggggatatt tttataatta ttttttttat agtttttaga tcttcttttt
     6061 tagagcgcct tgtaggcctt tatccatgct ggttctagag aaggtgttgt gacaaattgc
     6121 cctttcagtg tgacaaatca ccctcaaatg acagtcctgt ctgtgacaaa ttgcccttaa
     6181 ccctgtgaca aattgccctc agaagaagct gttttttcac aaagttatcc ctgcttattg
     6241 actctttttt atttagtgtg acaatctaaa aacttgtcac acttcacatg gatctgtcat
     6301 ggcggaaaca gcggttatca atcacaagaa acgtaaaaat agcccgcgaa tcgtccagtc
     6361 aaacgacctc actgaggcgg catatagtct ctcccgggat caaaaacgta tgctgtatct
     6421 gttcgttgac cagatcagaa aatctgatgg caccctacag gaacatgacg gtatctgcga
     6481 gatccatgtt gctaaatatg ctgaaatatt cggattgacc tctgcggaag ccagtaagga
     6541 tatacggcag gcattgaaga gtttcgcggg gaaggaagtg gttttttatc gccctgaaga
     6601 ggatgccggc gatgaaaaag gctatgaatc ttttccttgg tttatcaaac gtgcgcacag
     6661 tccatccaga gggctttaca gtgtacatat caacccatat ctcattccct tctttatcgg
     6721 gttacagaac cggtttacgc agtttcggct tagtgaaaca aaagaaatca ccaatccgta
     6781 tgccatgcgt ttatacgaat ccctgtgtca gtatcgtaag ccggatggct caggcatcgt
     6841 ctctctgaaa atcgactgga tcatagagcg ttaccagctg cctcaaagtt accagcgtat
     6901 gcctgacttc cgccgccgct tcctgcaggt ctgtgttaat gagatcaaca gcagaactcc
     6961 aatgcgcctc tcatacattg agaaaaagaa aggccgccag acgactcata tcgtattttc
     7021 cttccgcgat atcacttcca tgacgacagg atagtctgag ggttatctgt cacagatttg
     7081 agggtggttc gtcacatttg ttctgaccta ctgagggtaa tttgtcacag ttttgctgtt
     7141 tccttcagcc tgcatggatt ttctcatact ttttgaactg taatttttaa ggaagccaaa
     7201 tttgagggca gtttgtcaca gttgatttcc ttctctttcc cttcgtcatg tgacttgata
     7261 tcgggggtta gttcgtcatc attgatgagg gttgattatc acagtttatt actctgaatt
     7321 ggctatccgc gtgtgtacct ctacctggag tttttcccac ggtggatatt tcttcttgcg
     7381 ctgagcgtaa gagctatctg acagaacagt tcttctttgc ttcctcgcca gttcgctcgc
     7441 tatgctcggt tacacggctg cggcgagcgc tagtgataat aagtgactga ggtatgtgct
     7501 cttcttatct ccttttgtag tgttgctctt attttaaaca actttgcggt tttttgatga
     7561 ctttgcgatt ttgttgttgc tttgcagtaa attgcaagat ttaa
//
