LOCUS       sequence-440605-        7625 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To track the natural PhoB promoter's transcriptional dynamics.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7625)
  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 7625)
  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: PhoB-GFP
FEATURES             Location/Qualifiers
     source          1..7625
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    926..1176
                     /gene="incC"
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     CDS             complement(1179..1934)
                     /codon_start=1
                     /gene="repE"
                     /product="replication initiation protein for the bacterial 
                     F plasmid"
                     /label=repE
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     rep_origin      2025..2244
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     rep_origin      2312..2843
                     /label=oriV
                     /note="incP origin of replication"
     rep_origin      2320..2934
                     /label=oriV
                     /note="origin of replication for the bacterial F plasmid"
     promoter        3792..3894
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             3895..4554
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(3961..3980)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      complement(4689..4783)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(4846..5559)
                     /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(4886..4906)
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     5501..5529
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     RBS             5565..5573
                     /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      5835..5921
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(6079..7050)
                     /codon_start=1
                     /gene="sopB"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopB
                     /translation="MKRAPVIPKHTLNTQPVEDTSLSTPAAPMVDSLIARVGVMARGNA
                     ITLPVCGRDVKFTLEVLRGDSVEKTSRVWSGNERDQELLTEDALDDLIPSFLLTGQQTP
                     AFGRRVSGVIEIADGSRRRKAAALTESDYRVLVGELDDEQMAALSRLGNDYRPTSAYER
                     GQRYASRLQNEFAGNISALADAENISRKIITRCINTAKLPKSVVALFSHPGELSARSGD
                     ALQKAFTDKEELLKQQASNLHEQKKAGVIFEAEEVITLLTSVLKTSSASRTSLSSRHQF
                     APGATVLYKGDKMVLNLDRSRVPTECIEKIEAILKELEKPAP"
     CDS             complement(join(7050..7625,1..600))
                     /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 acaggaagga ataatgtcaa gccccggcca gcaagtgggc tttattgcat aagtgacatc
       61 gtccttttcc ccaagataga aaggcaggag agtgtcttct gcatgaatat gaagatctgg
      121 tacccatccg tgatacattg aggctgttcc ctgggggtcg ttaccttcca cgagcaaaac
      181 acgtagcccc ttcagagcca gatcctgagc aagatgaaca gaaactgagg ttttgtaaac
      241 gccaccttta tgggcagcaa ccccgatcac cggtggaaat acgtcttcag cacgtcgcaa
      301 tcgcgtacca aacacatcac gcatatgatt aatttgttca attgtataac caacacgttg
      361 ctcaacccgt cctcgaattt ccatatccgg gtgcggtagt cgccctgctt tctcggcatc
      421 tctgatagcc tgagaagaaa ccccaactaa atccgctgct tcacctattc tccagcgccg
      481 ggttattttc ctcgcttccg ggctgtcatc attaaactgt gcaatggcga tagccttcgt
      541 catttcatga ccagcgttta tgcactggtt aagtgtttcc atgagtttca ttctgaacat
      601 cctttaatca ttgctttgcg tttttttatt aaatcttgca atttactgca aagcaacaac
      661 aaaatcgcaa agtcatcaaa aaaccgcaaa gttgtttaaa ataagagcaa cactacaaaa
      721 ggagataaga agagcacata cctcagtcac ttattatcac tagcgctcgc cgcagccgtg
      781 taaccgagca tagcgagcga actggcgagg aagcaaagaa gaactgttct gtcagatagc
      841 tcttacgctc agcgcaagaa gaaatatcca ccgtgggaaa aactccaggt agaggtacac
      901 acgcggatag ccaattcaga gtaataaact gtgataatca accctcatca atgatgacga
      961 actaaccccc gatatcaagt cacatgacga agggaaagag aaggaaatca actgtgacaa
     1021 actgccctca aatttggctt ccttaaaaat tacagttcaa aaagtatgag aaaatccatg
     1081 caggctgaag gaaacagcaa aactgtgaca aattaccctc agtaggtcag aacaaatgtg
     1141 acgaaccacc ctcaaatctg tgacagataa ccctcagact atcctgtcgt catggaagtg
     1201 atatcgcgga aggaaaatac gatatgagtc gtctggcggc ctttcttttt ctcaatgtat
     1261 gagaggcgca ttggagttct gctgttgatc tcattaacac agacctgcag gaagcggcgg
     1321 cggaagtcag gcatacgctg gtaactttga ggcagctggt aacgctctat gatccagtcg
     1381 attttcagag agacgatgcc tgagccatcc ggcttacgat actgacacag ggattcgtat
     1441 aaacgcatgg catacggatt ggtgatttct tttgtttcac taagccgaaa ctgcgtaaac
     1501 cggttctgta acccgataaa gaagggaatg agatatgggt tgatatgtac actgtaaagc
     1561 cctctggatg gactgtgcgc acgtttgata aaccaaggaa aagattcata gcctttttca
     1621 tcgccggcat cctcttcagg gcgataaaaa accacttcct tccccgcgaa actcttcaat
     1681 gcctgccgta tatccttact ggcttccgca gaggtcaatc cgaatatttc agcatattta
     1741 gcaacatgga tctcgcagat accgtcatgt tcctgtaggg tgccatcaga ttttctgatc
     1801 tggtcaacga acagatacag catacgtttt tgatcccggg agagactata tgccgcctca
     1861 gtgaggtcgt ttgactggac gattcgcggg ctatttttac gtttcttgtg attgataacc
     1921 gctgtttccg ccatgacaga tccatgtgaa gtgtgacaag tttttagatt gtcacactaa
     1981 ataaaaaaga gtcaataagc agggataact ttgtgaaaaa acagcttctt ctgagggcaa
     2041 tttgtcacag ggttaagggc aatttgtcac agacaggact gtcatttgag ggtgatttgt
     2101 cacactgaaa gggcaatttg tcacaacacc ttctctagaa ccagcatgga taaaggccta
     2161 caaggcgctc taaaaaagaa gatctaaaaa ctataaaaaa aataattata aaaatatccc
     2221 cgtggataag tggataaccc caagggaagt tttttcaggc atcgtgtgta agcagaatat
     2281 ataagtgctg ttccctggtg cttcctcgct cactcgaccg ggagggttcg agaagggggg
     2341 gcacccccct tcggcgtgcg cggtcacgcg cacagggcgc agccctggtt aaaaacaagg
     2401 tttataaata ttggtttaaa agcaggttaa aagacaggtt agcggtggcc gaaaaacggg
     2461 cggaaaccct tgcaaatgct ggattttctg cctgtggaca gcccctcaaa tgtcaatagg
     2521 tgcgcccctc atctgtcagc actctgcccc tcaagtgtca aggatcgcgc ccctcatctg
     2581 tcagtagtcg cgcccctcaa gtgtcaatac cgcagggcac ttatccccag gcttgtccac
     2641 atcatctgtg ggaaactcgc gtaaaatcag gcgttttcgc cgatttgcga ggctggccag
     2701 ctccacgtcg ccggccgaaa tcgagcctgc ccctcatctg tcaacgccgc gccgggtgag
     2761 tcggcccctc aagtgtcaac gtccgcccct catctgtcag tgagggccaa gttttccgcg
     2821 aggtatccac aacgccggcg gccggccgcg gtgtctcgca cacggcttcg acggcgtttc
     2881 tggcgcgttt gcagggccat agacggccgc cagcccagcg gcgagggcaa ccagccgagg
     2941 gcttcgccct gtcgctcgac tgcggcgagc actactggct gtaaaaggac agaccacatc
     3001 atggttctgt gttcattagg ttgttctgtc cattgctgac ataatccgct ccacttcaac
     3061 gtaacaccgc acgaagattt ctattgttcc tgaaggcata ttcaaatcgt tttcgttacc
     3121 gcttgctggc atcatgacag aacactactt cctataaacg ctacacaggc tcctgagatt
     3181 aataatgcgg atctctacga taatgggaga ttttcccgac tgtttcgttc gcttctcagt
     3241 ggataacagc cagcttctct gtttaacaga caaaaacagc atatccactc agttccacat
     3301 ttccatataa aggccaaggc atttattctc aggataattg tttcagcatc gcaaccgcat
     3361 cagactccgg catcgcaaac tgcacccggt gccgggcagc cacatccagc gcaaaaacct
     3421 tcgtgtagac ttccgttgaa ctgatggact tatgtcccat caggctttgc agaactttca
     3481 gcggtatacc ggcatacagc atgtgcatcg cataggaatg gcggaacgta tgtggtgtga
     3541 ccggaacaga gaacgtcaca ccgtcagcag cagcggcggc aaccgcctcc ccaatccagg
     3601 tcctgaccgt tctgtccgtc acttcccaga tccgcgcttt ctctgtcctt cctgtgcgac
     3661 ggttacgccg ctccatgagc ttatcgcgaa taaatacctg tgacggaaga tcacttcgca
     3721 gaataaataa atcctggtgt ccctgttgat accgggaagc cctgggccaa cttttggcga
     3781 aaatgagacg ttgatcggca cgtaagaggt tccaactttc accataatga aataagatca
     3841 ctaccgggcg tattttttga gttatcgaga ttttcaggag ctaaggaagc taaaatggag
     3901 aaaaaaatca ctggatatac caccgttgat atatcccaat ggcatcgtaa agaacatttt
     3961 gaggcatttc agtcagttgc tcaatgtacc tataaccaga ccgttcagct ggatattacg
     4021 gcctttttaa agaccgtaaa gaaaaataag cacaagtttt atccggcctt tattcacatt
     4081 cttgcccgcc tgatgaatgc tcatccggaa tttcgtatgg caatgaaaga cggtgagctg
     4141 gtgatatggg atagtgttca cccttgttac accgttttcc atgagcaaac tgaaacgttt
     4201 tcatcgctct ggagtgaata ccacgacgat ttccggcagt ttctacacat atattcgcaa
     4261 gatgtggcgt gttacggtga aaacctggcc tatttcccta aagggtttat tgagaatatg
     4321 tttttcgtct cagccaatcc ctgggtgagt ttcaccagtt ttgatttaaa cgtggccaat
     4381 atggacaact tcttcgcccc cgttttcacg atgggcaaat attatacgca aggcgacaag
     4441 gtgctgatgc cgctggcgat tcaggttcat catgccgttt gtgatggctt ccatgtcggc
     4501 agaatgctta atgaattaca acagtactgc gatgagtggc agggcggggc gtaatttttt
     4561 taaggcagtt attggtgccc ttaaacgcct ggttgctacg cctgaataag tgataataag
     4621 cggatgaatg gcagaaattc gatgataagc tgtcaaacat gagaattggt ctcgacgagt
     4681 atgcttggat tctcaccaat aaaaaacgcc cggcggcaac cgagcgttct gaacaaatcc
     4741 agatggagtt ctgaggtcat tactggatct atcaacagga gtccaagcga cgtctgtgca
     4801 agtactactg ttctgcagtc acttgaattc gatacccagc tgttatttgt atagttcatc
     4861 catgccatgt gtaatcccag cagcggttac aaactcaaga aggaccatgt ggtctctctt
     4921 ttcgttggga tctttcgaaa gggcagattg tgtggacagg taatggttgt ctggtaaaag
     4981 gacagggcca tcgccaatcg gagtattttg ttgataatgg tctgctagtt gaacgcttcc
     5041 atcttcaatg ttgtgtctaa ttttgaagtt aactttgatt ccattctttt gtttgtctgc
     5101 catgatgtat acattgtgtg agttatagtt gtattccaat ttgtgtccaa gaatgtttcc
     5161 atcttcttta aaatcaatac cttttaactc gattctatta acaagggtat caccttcaaa
     5221 cttgacttca gcacgtgtct tgtagttccc gtcatctttg aaaaatatag ttctttcctg
     5281 tacgtaacct tcgggcatgg cactcttgaa aaagtcatgc tgtttcatgt gatctgggta
     5341 tctcgcaaag cattgaacac cataaccgaa agtagtgaca agtgttggcc acggaacagg
     5401 tagttttcca gtagtgcaaa taaatttaag ggtaagtttt ccgtatgttg catcaccttc
     5461 accctctcca ctgacagaaa atttgtgccc attaacatca ccatctaatt caacaagaat
     5521 tgggacaact ccagtgaaaa gttcttctcc tttacgcata ttatacctcc ttaatttagt
     5581 gtcatctaca actggcatga tttgccctgt tgtaataaat aggttgcgat cattaatgcg
     5641 acgtcattat gcgtcagatt tatgacagat ttatgaaaag ctcgtcgcac atatcttcag
     5701 gttattgatt tccgtggcgc agaaaaaagc aaatggcaca tctgtttggg tataatcgcg
     5761 cccatgcttt ttcgccaggg aaccgttcct agggaccgtc tcgagagaat caatattaat
     5821 ccaacgcgtg gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta
     5881 tctgttgttt gtcggtgaac gctctcctga gtaggacaaa tccgccgccc tagacttagg
     5941 cgttcggctg cggcgagcgg tatcagctca ctcaaagaag tggggcccag agagaatatt
     6001 caggccagtt atgctttctg gcctgtaaca aaggacatta agtaaagaca gataaacgta
     6061 gactaaaacg tggtcgcatc agggtgctgg cttttcaagt tccttaagaa tggcctcaat
     6121 tttctctata cactcagttg gaacacggga cctgtccagg ttaagcacca ttttatcgcc
     6181 cttatacaat actgtcgctc caggagcaaa ctgatgtcgt gagcttaaac tagttcttga
     6241 tgcagatgac gttttaagca cagaagttaa aagagtgata acttcttcag cttcaaatat
     6301 caccccagct tttttctgct catgaaggtt agatgcctgc tgcttaagta attcctcttt
     6361 atctgtaaag gctttttgaa gtgcatcacc tgaccgggca gatagttcac cggggtgaga
     6421 aaaaagagca acaactgatt taggcaattt ggcggtgttg atacagcggg taataatctt
     6481 acgtgaaata ttttccgcat cagccagcgc agaaatattt ccagcaaatt cattctgcaa
     6541 tcggcttgca taacgctgac cacgttcata agcacttgtt gggcgataat cgttacccaa
     6601 tctggataat gcagccatct gctcatcatc cagctcgcca accagaacac gataatcact
     6661 ttcggtaagt gcagcagctt tacgacggcg actcccatcg gcaatttcta tgacaccaga
     6721 tactcttcga ccgaacgccg gtgtctgttg accagtcagt agaaaagaag ggatgagatc
     6781 atccagtgcg tcctcagtaa gcagctcctg gtcacgttca ttacctgacc atacccgaga
     6841 ggtcttctca acactatcac cccggagcac ttcaagagta aacttcacat cccgaccaca
     6901 tacaggcaaa gtaatggcat taccgcgagc cattactcct acgcgcgcaa ttaacgaatc
     6961 caccatcggg gcagctggtg tcgataacga agtatcttca accggttgag tattgagcgt
     7021 atgttttgga ataacaggcg cacgcttcat tatctaatct cccagcgtgg tttaatcaga
     7081 cgatcgaaaa tttcattgca gacaggttcc caaatagaaa gagcatttct ccaggcacca
     7141 gttgaagagc gttgatcaat ggcctgttca aaaacagttc tcatccggat ctgaccttta
     7201 ccaacttcat ccgtttcacg tacaacattt tttagaacca tgcttcccca ggcatcccga
     7261 atttgctcct ccatccacgg ggactgagag ccattactat tgctgtattt ggtaagcaaa
     7321 atacgtacat caggctcgaa ccctttaaga tcaacgttct tgagcagatc acgaagcata
     7381 tcgaaaaact gcagtgcgga ggtgtagtca aacaactcag caggcgtggg aacaatcagc
     7441 acatcagcag cacatacgac attaatcgtg ccgataccca ggttaggcgc gctgtcaata
     7501 actatgacat catagtcatg agcaacagtt tcaatggcca gtcggagcat caggtgtgga
     7561 tcggtgggca gtttaccttc atcaaatttg cccattaact cagtttcaat acggtgcaga
     7621 gccag
//
