LOCUS       sequence-436627-        7980 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To track the natural Lrp promoter's transcriptional dynamics.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7980)
  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 7980)
  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: Lrp-GFP
FEATURES             Location/Qualifiers
     source          1..7980
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(851..2026)
                     /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    2352..2602
                     /gene="incC"
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     CDS             complement(2605..3360)
                     /codon_start=1
                     /gene="repE"
                     /product="replication initiation protein for the bacterial 
                     F plasmid"
                     /label=repE
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     rep_origin      3451..3670
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     rep_origin      3738..4269
                     /label=oriV
                     /note="incP origin of replication"
     rep_origin      3746..4360
                     /label=oriV
                     /note="origin of replication for the bacterial F plasmid"
     promoter        5218..5320
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             5321..5980
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(5387..5406)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      complement(6115..6209)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(6272..6985)
                     /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(6312..6332)
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     6927..6955
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     RBS             6991..6999
                     /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      7616..7702
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(join(7860..7980,1..851))
                     /codon_start=1
                     /gene="sopB"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopB
                     /translation="MKRAPVIPKHTLNTQPVEDTSLSTPAAPMVDSLIARVGVMARGNA
                     ITLPVCGRDVKFTLEVLRGDSVEKTSRVWSGNERDQELLTEDALDDLIPSFLLTGQQTP
                     AFGRRVSGVIEIADGSRRRKAAALTESDYRVLVGELDDEQMAALSRLGNDYRPTSAYER
                     GQRYASRLQNEFAGNISALADAENISRKIITRCINTAKLPKSVVALFSHPGELSARSGD
                     ALQKAFTDKEELLKQQASNLHEQKKAGVIFEAEEVITLLTSVLKTSSASRTSLSSRHQF
                     APGATVLYKGDKMVLNLDRSRVPTECIEKIEAILKELEKPAP"
ORIGIN
        1 ccaggagcaa actgatgtcg tgagcttaaa ctagttcttg atgcagatga cgttttaagc
       61 acagaagtta aaagagtgat aacttcttca gcttcaaata tcaccccagc ttttttctgc
      121 tcatgaaggt tagatgcctg ctgcttaagt aattcctctt tatctgtaaa ggctttttga
      181 agtgcatcac ctgaccgggc agatagttca ccggggtgag aaaaaagagc aacaactgat
      241 ttaggcaatt tggcggtgtt gatacagcgg gtaataatct tacgtgaaat attttccgca
      301 tcagccagcg cagaaatatt tccagcaaat tcattctgca atcggcttgc ataacgctga
      361 ccacgttcat aagcacttgt tgggcgataa tcgttaccca atctggataa tgcagccatc
      421 tgctcatcat ccagctcgcc aaccagaaca cgataatcac tttcggtaag tgcagcagct
      481 ttacgacggc gactcccatc ggcaatttct atgacaccag atactcttcg accgaacgcc
      541 ggtgtctgtt gaccagtcag tagaaaagaa gggatgagat catccagtgc gtcctcagta
      601 agcagctcct ggtcacgttc attacctgac catacccgag aggtcttctc aacactatca
      661 ccccggagca cttcaagagt aaacttcaca tcccgaccac atacaggcaa agtaatggca
      721 ttaccgcgag ccattactcc tacgcgcgca attaacgaat ccaccatcgg ggcagctggt
      781 gtcgataacg aagtatcttc aaccggttga gtattgagcg tatgttttgg aataacaggc
      841 gcacgcttca ttatctaatc tcccagcgtg gtttaatcag acgatcgaaa atttcattgc
      901 agacaggttc ccaaatagaa agagcatttc tccaggcacc agttgaagag cgttgatcaa
      961 tggcctgttc aaaaacagtt ctcatccgga tctgaccttt accaacttca tccgtttcac
     1021 gtacaacatt ttttagaacc atgcttcccc aggcatcccg aatttgctcc tccatccacg
     1081 gggactgaga gccattacta ttgctgtatt tggtaagcaa aatacgtaca tcaggctcga
     1141 accctttaag atcaacgttc ttgagcagat cacgaagcat atcgaaaaac tgcagtgcgg
     1201 aggtgtagtc aaacaactca gcaggcgtgg gaacaatcag cacatcagca gcacatacga
     1261 cattaatcgt gccgataccc aggttaggcg cgctgtcaat aactatgaca tcatagtcat
     1321 gagcaacagt ttcaatggcc agtcggagca tcaggtgtgg atcggtgggc agtttacctt
     1381 catcaaattt gcccattaac tcagtttcaa tacggtgcag agccagacag gaaggaataa
     1441 tgtcaagccc cggccagcaa gtgggcttta ttgcataagt gacatcgtcc ttttccccaa
     1501 gatagaaagg caggagagtg tcttctgcat gaatatgaag atctggtacc catccgtgat
     1561 acattgaggc tgttccctgg gggtcgttac cttccacgag caaaacacgt agccccttca
     1621 gagccagatc ctgagcaaga tgaacagaaa ctgaggtttt gtaaacgcca cctttatggg
     1681 cagcaacccc gatcaccggt ggaaatacgt cttcagcacg tcgcaatcgc gtaccaaaca
     1741 catcacgcat atgattaatt tgttcaattg tataaccaac acgttgctca acccgtcctc
     1801 gaatttccat atccgggtgc ggtagtcgcc ctgctttctc ggcatctctg atagcctgag
     1861 aagaaacccc aactaaatcc gctgcttcac ctattctcca gcgccgggtt attttcctcg
     1921 cttccgggct gtcatcatta aactgtgcaa tggcgatagc cttcgtcatt tcatgaccag
     1981 cgtttatgca ctggttaagt gtttccatga gtttcattct gaacatcctt taatcattgc
     2041 tttgcgtttt tttattaaat cttgcaattt actgcaaagc aacaacaaaa tcgcaaagtc
     2101 atcaaaaaac cgcaaagttg tttaaaataa gagcaacact acaaaaggag ataagaagag
     2161 cacatacctc agtcacttat tatcactagc gctcgccgca gccgtgtaac cgagcatagc
     2221 gagcgaactg gcgaggaagc aaagaagaac tgttctgtca gatagctctt acgctcagcg
     2281 caagaagaaa tatccaccgt gggaaaaact ccaggtagag gtacacacgc ggatagccaa
     2341 ttcagagtaa taaactgtga taatcaaccc tcatcaatga tgacgaacta acccccgata
     2401 tcaagtcaca tgacgaaggg aaagagaagg aaatcaactg tgacaaactg ccctcaaatt
     2461 tggcttcctt aaaaattaca gttcaaaaag tatgagaaaa tccatgcagg ctgaaggaaa
     2521 cagcaaaact gtgacaaatt accctcagta ggtcagaaca aatgtgacga accaccctca
     2581 aatctgtgac agataaccct cagactatcc tgtcgtcatg gaagtgatat cgcggaagga
     2641 aaatacgata tgagtcgtct ggcggccttt ctttttctca atgtatgaga ggcgcattgg
     2701 agttctgctg ttgatctcat taacacagac ctgcaggaag cggcggcgga agtcaggcat
     2761 acgctggtaa ctttgaggca gctggtaacg ctctatgatc cagtcgattt tcagagagac
     2821 gatgcctgag ccatccggct tacgatactg acacagggat tcgtataaac gcatggcata
     2881 cggattggtg atttcttttg tttcactaag ccgaaactgc gtaaaccggt tctgtaaccc
     2941 gataaagaag ggaatgagat atgggttgat atgtacactg taaagccctc tggatggact
     3001 gtgcgcacgt ttgataaacc aaggaaaaga ttcatagcct ttttcatcgc cggcatcctc
     3061 ttcagggcga taaaaaacca cttccttccc cgcgaaactc ttcaatgcct gccgtatatc
     3121 cttactggct tccgcagagg tcaatccgaa tatttcagca tatttagcaa catggatctc
     3181 gcagataccg tcatgttcct gtagggtgcc atcagatttt ctgatctggt caacgaacag
     3241 atacagcata cgtttttgat cccgggagag actatatgcc gcctcagtga ggtcgtttga
     3301 ctggacgatt cgcgggctat ttttacgttt cttgtgattg ataaccgctg tttccgccat
     3361 gacagatcca tgtgaagtgt gacaagtttt tagattgtca cactaaataa aaaagagtca
     3421 ataagcaggg ataactttgt gaaaaaacag cttcttctga gggcaatttg tcacagggtt
     3481 aagggcaatt tgtcacagac aggactgtca tttgagggtg atttgtcaca ctgaaagggc
     3541 aatttgtcac aacaccttct ctagaaccag catggataaa ggcctacaag gcgctctaaa
     3601 aaagaagatc taaaaactat aaaaaaaata attataaaaa tatccccgtg gataagtgga
     3661 taaccccaag ggaagttttt tcaggcatcg tgtgtaagca gaatatataa gtgctgttcc
     3721 ctggtgcttc ctcgctcact cgaccgggag ggttcgagaa gggggggcac cccccttcgg
     3781 cgtgcgcggt cacgcgcaca gggcgcagcc ctggttaaaa acaaggttta taaatattgg
     3841 tttaaaagca ggttaaaaga caggttagcg gtggccgaaa aacgggcgga aacccttgca
     3901 aatgctggat tttctgcctg tggacagccc ctcaaatgtc aataggtgcg cccctcatct
     3961 gtcagcactc tgcccctcaa gtgtcaagga tcgcgcccct catctgtcag tagtcgcgcc
     4021 cctcaagtgt caataccgca gggcacttat ccccaggctt gtccacatca tctgtgggaa
     4081 actcgcgtaa aatcaggcgt tttcgccgat ttgcgaggct ggccagctcc acgtcgccgg
     4141 ccgaaatcga gcctgcccct catctgtcaa cgccgcgccg ggtgagtcgg cccctcaagt
     4201 gtcaacgtcc gcccctcatc tgtcagtgag ggccaagttt tccgcgaggt atccacaacg
     4261 ccggcggccg gccgcggtgt ctcgcacacg gcttcgacgg cgtttctggc gcgtttgcag
     4321 ggccatagac ggccgccagc ccagcggcga gggcaaccag ccgagggctt cgccctgtcg
     4381 ctcgactgcg gcgagcacta ctggctgtaa aaggacagac cacatcatgg ttctgtgttc
     4441 attaggttgt tctgtccatt gctgacataa tccgctccac ttcaacgtaa caccgcacga
     4501 agatttctat tgttcctgaa ggcatattca aatcgttttc gttaccgctt gctggcatca
     4561 tgacagaaca ctacttccta taaacgctac acaggctcct gagattaata atgcggatct
     4621 ctacgataat gggagatttt cccgactgtt tcgttcgctt ctcagtggat aacagccagc
     4681 ttctctgttt aacagacaaa aacagcatat ccactcagtt ccacatttcc atataaaggc
     4741 caaggcattt attctcagga taattgtttc agcatcgcaa ccgcatcaga ctccggcatc
     4801 gcaaactgca cccggtgccg ggcagccaca tccagcgcaa aaaccttcgt gtagacttcc
     4861 gttgaactga tggacttatg tcccatcagg ctttgcagaa ctttcagcgg tataccggca
     4921 tacagcatgt gcatcgcata ggaatggcgg aacgtatgtg gtgtgaccgg aacagagaac
     4981 gtcacaccgt cagcagcagc ggcggcaacc gcctccccaa tccaggtcct gaccgttctg
     5041 tccgtcactt cccagatccg cgctttctct gtccttcctg tgcgacggtt acgccgctcc
     5101 atgagcttat cgcgaataaa tacctgtgac ggaagatcac ttcgcagaat aaataaatcc
     5161 tggtgtccct gttgataccg ggaagccctg ggccaacttt tggcgaaaat gagacgttga
     5221 tcggcacgta agaggttcca actttcacca taatgaaata agatcactac cgggcgtatt
     5281 ttttgagtta tcgagatttt caggagctaa ggaagctaaa atggagaaaa aaatcactgg
     5341 atataccacc gttgatatat cccaatggca tcgtaaagaa cattttgagg catttcagtc
     5401 agttgctcaa tgtacctata accagaccgt tcagctggat attacggcct ttttaaagac
     5461 cgtaaagaaa aataagcaca agttttatcc ggcctttatt cacattcttg cccgcctgat
     5521 gaatgctcat ccggaatttc gtatggcaat gaaagacggt gagctggtga tatgggatag
     5581 tgttcaccct tgttacaccg ttttccatga gcaaactgaa acgttttcat cgctctggag
     5641 tgaataccac gacgatttcc ggcagtttct acacatatat tcgcaagatg tggcgtgtta
     5701 cggtgaaaac ctggcctatt tccctaaagg gtttattgag aatatgtttt tcgtctcagc
     5761 caatccctgg gtgagtttca ccagttttga tttaaacgtg gccaatatgg acaacttctt
     5821 cgcccccgtt ttcacgatgg gcaaatatta tacgcaaggc gacaaggtgc tgatgccgct
     5881 ggcgattcag gttcatcatg ccgtttgtga tggcttccat gtcggcagaa tgcttaatga
     5941 attacaacag tactgcgatg agtggcaggg cggggcgtaa tttttttaag gcagttattg
     6001 gtgcccttaa acgcctggtt gctacgcctg aataagtgat aataagcgga tgaatggcag
     6061 aaattcgatg ataagctgtc aaacatgaga attggtctcg acgagtatgc ttggattctc
     6121 accaataaaa aacgcccggc ggcaaccgag cgttctgaac aaatccagat ggagttctga
     6181 ggtcattact ggatctatca acaggagtcc aagcgacgtc tgtgcaagta ctactgttct
     6241 gcagtcactt gaattcgata cccagctgtt atttgtatag ttcatccatg ccatgtgtaa
     6301 tcccagcagc ggttacaaac tcaagaagga ccatgtggtc tctcttttcg ttgggatctt
     6361 tcgaaagggc agattgtgtg gacaggtaat ggttgtctgg taaaaggaca gggccatcgc
     6421 caatcggagt attttgttga taatggtctg ctagttgaac gcttccatct tcaatgttgt
     6481 gtctaatttt gaagttaact ttgattccat tcttttgttt gtctgccatg atgtatacat
     6541 tgtgtgagtt atagttgtat tccaatttgt gtccaagaat gtttccatct tctttaaaat
     6601 caataccttt taactcgatt ctattaacaa gggtatcacc ttcaaacttg acttcagcac
     6661 gtgtcttgta gttcccgtca tctttgaaaa atatagttct ttcctgtacg taaccttcgg
     6721 gcatggcact cttgaaaaag tcatgctgtt tcatgtgatc tgggtatctc gcaaagcatt
     6781 gaacaccata accgaaagta gtgacaagtg ttggccacgg aacaggtagt tttccagtag
     6841 tgcaaataaa tttaagggta agttttccgt atgttgcatc accttcaccc tctccactga
     6901 cagaaaattt gtgcccatta acatcaccat ctaattcaac aagaattggg acaactccag
     6961 tgaaaagttc ttctccttta cgcatattat acctccttaa tttagtgtca tctacaactg
     7021 gcattattat tgtctctctg tattccttcc ctactcctgt ctgaccctga aacatcactg
     7081 ttcagagtcg ctacccgtca cgatagaccg aaatcatgaa gattcacggc gcgcaatgcc
     7141 tgggtctacg gtgagaagac cgttgccttg tggcagctat cgacatcacg aatggcgtct
     7201 gtccacgcca tacgtagatt tcactagcac ggtaaacatg gtatttacat gcatggttat
     7261 gcgcatataa gaatactgtt ttttttcttc ctcgaatgtt ttcgcaaaac accaggggat
     7321 tgtcaaagca aaacatcgat ttttagtaca acatgccaga tattcattaa caggggcgtt
     7381 tattcatcat ttaatcggct tataaagcgg gaaatttaag tagcagaggt gataatgacg
     7441 ggagaatttg acctgcttat cgatgctgcc gattgcacaa attgttaaca aaatcgtcat
     7501 actctttttt tacgtctgta aattccctac aatcctgccc attgtctgcc aacaactatg
     7561 gggatctccc tagggaccgt ctcgagagaa tcaatattaa tccaacgcgt ggcatcaaat
     7621 aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa
     7681 cgctctcctg agtaggacaa atccgccgcc ctagacttag gcgttcggct gcggcgagcg
     7741 gtatcagctc actcaaagaa gtggggccca gagagaatat tcaggccagt tatgctttct
     7801 ggcctgtaac aaaggacatt aagtaaagac agataaacgt agactaaaac gtggtcgcat
     7861 cagggtgctg gcttttcaag ttccttaaga atggcctcaa ttttctctat acactcagtt
     7921 ggaacacggg acctgtccag gttaagcacc attttatcgc ccttatacaa tactgtcgct
//
