LOCUS       sequence-251809-        8866 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of torS under Ptac and torT under PD59_luxR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8866)
  AUTHORS   Schmidl SR, Ekness F, Sofjan K, Daeffler KN, Brink KR, Landry BP, 
            Gerhardt KP, Dyulgyarov N, Sheth RU, Tabor JJ
  TITLE     Rewiring bacterial two-component systems by modular DNA-binding 
            domain swapping.
  JOURNAL   Nat Chem Biol. 2019 Jul;15(7):690-698. doi: 
            10.1038/s41589-019-0286-6. Epub 2019 May 20.
  PUBMED    31110305
REFERENCE   2  (bases 1 to 8866)
  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"; doi: 
            "10.1038/s41589-019-0286-6"; journalName: "Nat Chem Biol"; date: 
            "2019-07"; volume: "15"; issue: "7"; pages: "690-698"
COMMENT     Sequence Label: pKD251
FEATURES             Location/Qualifiers
     source          1..8866
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(105..110,111..126,127..133)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    141..157
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     terminator      2966..3037
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3053..3080
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     3244..3262
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(3266..3309)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     promoter        3347..3381
                     /label=BBa_J23107
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.36"
     RBS             3390..3401
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      4205..4276
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4292..4319
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      5503..5574
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5590..5617
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             5817..6608
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     terminator      6620..6714
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      6828..7373
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     7274..7293
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7490..7507
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        7546..7623
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             7624..8706
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(7643..7662)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     terminator      8712..8798
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 taaatcactg cataattcgt gtcgctcaag gcgcactccc gttctggata atgttttttg
       61 cgccgacatc ataacggttc tggctaatat tctgaaatga gctgttgaca attaatcatc
      121 ggctcgtata atgtgtggaa ttgtgagcgg ataacaattt cacacagtcc cctcgccaca
      181 acaagtataa cgctttatac atgaatttaa ccctgacccg aagactctgg atgggctttg
      241 ccctgatggc gctgttaacc ctgaccagta ccctggtggg atggtacaac ctgcgcttta
      301 tcagccaggt ggaaaaagac aacactcagg cattgattcc taccatgaat atggcgcgcc
      361 agttgagcga agccagcgcc tgggaacttt tcgccgcgca gaacctgacc agtgccgata
      421 acgaaaagat gtggcaggcg caggggcgaa tgctcaccgc acaaagcctg aagattaatg
      481 cgttgctgca agcgttacgg gaacaaggtt ttgataccac cgctattgaa caacaggagc
      541 aggagatctc ccgttcgtta cgtcagcaag gggaactggt ggggcagcgt ttgcaactgc
      601 gccagcaaca acagcaactc agtcagcaga tagtcgctgc cgccgatgag atcgcacgcc
      661 tggcgcaagg tcaggcgaat aatgcgacaa cttccgctgg agcgacccag gccgggattt
      721 acgatttgat cgaacaagat cagcgtcagg ctgctgaaag tgcactcgat cggctgattg
      781 atatcgatct tgagtatgtt aaccagatga atgaactgcg ccttagcgct ctgcgggtgc
      841 agcaaatggt gatgaatctg gggctggagc agatccagaa aaatgcacca acgctggaaa
      901 agcagctcaa taatgcggtg aaaattctgc aacgtcggca aatacgcatt gaagatccgg
      961 gtgttcgtgc gcaggtcgca acaacgttaa ctaccgttag ccaatatagc gatttgctgg
     1021 cgctgtatca gcaggacagt gaaatcagca atcacctaca aactctcgca caaaataaca
     1081 tcgcccagtt cgcgcagttt agtagcgaag tcagtcagct ggtcgacacc attgagctgc
     1141 gtaatcagca cggactggcg catctggaaa aagcgagtgc acgcgggcaa tacagcctgt
     1201 tattgctggg gatggtttca ctttgcgcac tgattctgat cctctggcgc gtggtttatc
     1261 gctcagtcac gcgtccactt gccgaacaaa cgcaggcgct gcaacggctg ctggacggtg
     1321 atattgactc ccctttcccg gaaaccgctg gcgtacggga gctggatacc atcgggcggc
     1381 tgatggatgc gtttcgcagc aatgttcatg cactgaatcg ccaccgcgaa cagctggcgg
     1441 cgcaggtcaa agcgcgtaca gctgaattgc aggaactggt gatagaacac cgacaggcac
     1501 gggcggaagc agaaaaagcc agccaggcaa aatcggcgtt tctggcggcg atgagccatg
     1561 agatccgcac accgctgtac ggtattctcg gcactgctca actgctggca gataaccccg
     1621 cacttaacgc ccagcgtgat gatttgcggg caattactga ctctggtgaa tcgttgctga
     1681 ccatcctcaa cgatattctc gattattccg ctatcgaagc aggtggcaag aatgtttcgg
     1741 tcagcgatga gccctttgaa ccgcgcccgc tgctggaaag taccctgcaa ttaatgagcg
     1801 gacgggtgaa aggtcgcccg attcgcctgg caacggcaat tgccgatgat atgccgtgcg
     1861 cgttaatggg cgatccacga cgtattcgtc aggttataac caacctgttg agcaacgccc
     1921 tgcgttttac tgacgaaggg tacattatcc tgcgtagtcg cactgatggc gagcaatggc
     1981 tggtcgaagt ggaagacagc ggctgcggta ttgatcccgc gaaactggca gaaatcttcc
     2041 agccatttgt gcaggtaagc ggcaaacgcg gcggcaccgg gctgggactg actatcagta
     2101 gccgtctggc ccaggcgatg ggcggcgaac tgagcgccac cagcacgccg gaggttggaa
     2161 gctgtttttg tttacgcttg ccgttacgtg ttgccacggc acccgtgcca aaaacagtca
     2221 atcaggcggt gcgtcttgac ggtttacgtt tgctgttaat tgaagataac ccgctaaccc
     2281 agcgaattac cattgagatg ctgaaaacca gtggtgcgca gattgttgct gttggcaatg
     2341 ccgcgcaggc tttagagaca ctgcaaaata gcgaaccgtt tgctgccgca ctggtggatt
     2401 ttgatctgcc ggatatcgac ggcattacgc ttgcccgaca actggcacag caatatccgt
     2461 cgctggtttt gattggcttt agcgcccatg tcattgacga aacgctgcgc cagcgtacca
     2521 gttcattatt ccgcgggatt atccctaaac cggtgccgcg tgaagtgctc ggtcagttac
     2581 tggcgcacta tctccaactg caagtcaata atgatcaatc gctggatgta tcgcaactca
     2641 atgaagatgc tcagttaatg gggacggaga agatccacga atggctggta ttatttacac
     2701 aacatgccct gccgcttctc gatgaaatcg acattgcccg cgccagtcag gatagcgaga
     2761 aaataaagcg tgccgcgcat cagctaaaaa gcagttgctc gagtctggga atgcatatcg
     2821 ccagccagct atgcgcacaa ctggagcagc agccattatc tgcccccctt ccacacgaag
     2881 aaattacacg cagtgttgcc gctctggaag catggttgca taagaaagac ctgaacgcga
     2941 tttaataata atctagacca ggcatcaaat aaaacgaaag gctcagtcga aagactgggc
     3001 ctttcgtttt atctgttgtt tgtcggtgaa cgctctctac tagagtcaca ctggctcacc
     3061 ttcgggtggg cctttctgcg tttatagtaa atcactgcat aattcgtgtc gctcaaggcg
     3121 cactcccgtt ctggataatg ttttttgcgc cgacatcata acggttctgg caaatattct
     3181 gaaatgagct ggttagctag tcacgtctaa gaaaccatta ttatcatgac attaacctat
     3241 aaaaataggc gtatcacgag gcagaatttc agataaaaaa aatccttagc tttcgctaag
     3301 gatgatttct gggtacccgg ggatcctcta gagtcgacct gcaggcttta cggctagctc
     3361 agccctaggt attatgctag ctactagaga aagaggagaa atactagatg aaaaacataa
     3421 atgccgacga cacatacaga ataattaata aaattaaagc ttgtagaagc aataatgata
     3481 ttaatcaatg cttatctgat atgactaaaa tggtacattg tgaatattat ttactcgcga
     3541 tcatttatcc tcattctatg gttaaatctg atatttcaat cctagataat taccctaaaa
     3601 aatggaggca atattatgat gacgctaatt taataaaata tgatcctata gtagattatt
     3661 ctaactccaa tcattcacca attaattgga atatatttga aaacaatgct gtaaataaaa
     3721 aatctccaaa tgtaattaaa gaagcgaaaa catcaggtct tatcactggg tttagtttcc
     3781 ctattcatac ggctaacaat ggcttcggaa tgcttagttt tgcacattca gaaaaagaca
     3841 actatataga tagtttattt ttacatgcgt gtatgaacat accattaatt gttccttctc
     3901 tagttgataa ttatcgaaaa ataaatatag caaataataa atcaaacaac gatttaacca
     3961 aaagagaaaa agaatgttta gcgtgggcat gcgaaggaaa aagctcttgg gatatttcaa
     4021 aaatattagg ttgcagtgag cgtactgtca ctttccattt aaccaatgcg caaatgaaac
     4081 tcaatacaac aaaccgctgc caaagtattt ctaaagcaat tttaacagga gcaattgatt
     4141 gcccatactt taaaaattaa taacactgat agtgctagtg tagatcacta ctagagccag
     4201 gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt
     4261 gtcggtgaac gctctctact agagtcacac tggctcacct tcgggtgggc ctttctgcgt
     4321 ttatatacaa ttgtttaaca taagtacctg taggatcgta caggtttact attttacctc
     4381 tggcggtgat aattcttgca acaaacaata ggtaaggcgt tacccaacac tatatatttt
     4441 acatcattcc gctttaattt caatgcgcgt actgctattt ttacttcttt cccttttcat
     4501 gttgccggca ttttcggctg ataacctgtt gcgctggcat gatgcgcagc atttcacggt
     4561 gcaagcctct acgccgctta aagccaaacg cgcatggaaa ctgtgcgcgc tttatcccag
     4621 cctgaaagat tcatattggt tatcgttgaa ctatggtatg caggaggctg ctcgccgcta
     4681 cggtgtggat ttaaaagtgc tggaggcagg cggctacagc cagttggcta cccagcaagc
     4741 acaaatcgac cagtgtaaac agtggggcgc agaggccatt ttgctcggta gtagcacgac
     4801 ctcatttccc gacctgcaaa agcaggtagc aagtctgccg gtgatcgaac tggtaaatgc
     4861 tattgatgct ccccaggtga aaagccgcgt tggtgtgccc tggtttcaga tgggctatca
     4921 accggggcga tatctggtgc aatgggcgca cggtaaacca ctgaatgtgc tgttgatgcc
     4981 cggacccgat aacgccgggg gcagtaagga gatggtcgag ggttttcgcg cagccattgc
     5041 cggaagcccg gtgcgtattg ttgatattgc gcttggtgat aacgatattg aaatccagcg
     5101 taacctgttg caggagatgc tggaacgcca tccagaaatc gacgtcgttg ccggaacggc
     5161 cattgcggca gaggcggcaa tgggggaagg gcgtaacctg aaaacgccgc ttaccgtggt
     5221 gtcgttttat ctttcacatc aggtgtatcg cgggctgaag cggggaagag tgattatggc
     5281 tgccagcgat caaatggtct ggcaggggga actggcggtt gagcaggcca tcaggcaatt
     5341 acaggggcaa tcggtttctg ataatgtcag cccaccgatt ttagttctga cgccgaaaaa
     5401 tgccgaccgt gaacatattc gccgctcgct gtcaccaggg ggatttcgtc cggtctattt
     5461 ttatcagcac acatcagcgg ctaagaaata ataaccaggc atcaaataaa acgaaaggct
     5521 cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctctactag
     5581 agtcacactg gctcaccttc gggtgggcct ttctgcgttt ataggtaccc ggggatcctc
     5641 tagagtcgac ctgcaggcat gcaagcttag caagcgaacc ggaattgcca gctggggcgc
     5701 cctctggtaa ggttgggaag ccctgcaaag taaactggat ggctttcttg ccgccaagga
     5761 tctgatggcg caggggatca agatctgatc aagagacagg atgaggatcg tttcgtatga
     5821 gggaagcggt gatcgccgaa gtatcgactc aactatcaga ggtagttggc gtcatcgagc
     5881 gccatctcga accgacgttg ctggccgtac atttgtacgg ctccgcagtg gatggcggcc
     5941 tgaagccaca cagtgatatt gatttgctgg ttacggtgac cgtaaggctt gatgaaacaa
     6001 cgcggcgagc tttgatcaac gaccttttgg aaacttcggc ttcccctgga gagagcgaga
     6061 ttctccgcgc tgtagaagtc accattgttg tgcacgacga catcattccg tggcgttatc
     6121 cagctaagcg cgaactgcaa tttggagaat ggcagcgcaa tgacattctt gcaggtatct
     6181 tcgagccagc cacgatcgac attgatctgg ctatcttgct gacaaaagca agagaacata
     6241 gcgttgcctt ggtaggtcca gcggcggagg aactctttga tccggttcct gaacaggatc
     6301 tatttgaggc gctaaatgaa accttaacgc tatggaactc gccgcccgac tgggctggcg
     6361 atgagcgaaa tgtagtgctt acgttgtccc gcatttggta cagcgcagta accggcaaaa
     6421 tcgcgccgaa ggatgtcgct gccgactggg caatggagcg cctgccggcc cagtatcagc
     6481 ccgtcatact tgaagctaga caggcttatc ttggacaaga agaagatcgc ttggcctcgc
     6541 gcgcagatca gttggaagaa tttgtccact acgtgaaagg cgagatcacc aaggtagtcg
     6601 gcaaataaga gctcgcttgg actcctgttg atagatccag taatgacctc agaactccat
     6661 ctggatttgt tcagaacgct cggttgccgc cgggcgtttt ttattggtga gaatccaagc
     6721 actagtaaca acttatatcg tatggggctg acttcaggtg ctacatttga agagataaat
     6781 tgcactgaaa tctagtaata ttttatctga ttaataagat gatcttcttg agatcgtttt
     6841 ggtctgcgcg taatctcttg ctctgaaaac gaaaaaaccg ccttgcaggg cggtttttcg
     6901 aaggttctct gagctaccaa ctctttgaac cgaggtaact ggcttggagg agcgcagtca
     6961 ccaaaacttg tcctttcagt ttagccttaa ccggcgcatg acttcaagac taactcctct
     7021 aaatcaatta ccagtggctg ctgccagtgg tgcttttgca tgtctttccg ggttggactc
     7081 aagacgatag ttaccggata aggcgcagcg gtcggactga acggggggtt cgtgcataca
     7141 gtccagcttg gagcgaactg cctacccgga actgagtgtc aggcgtggaa tgagacaaac
     7201 gcggccataa cagcggaatg acaccggtaa accgaaaggc aggaacagga gagcgcacga
     7261 gggagccgcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccaccact
     7321 gatttgagcg tcagatttcg tgatgcttgt caggggggcg gagcctatgg aaaaacggct
     7381 ttgccgcggc cctctcactt ccctgttaag tatcttcctg gcatcttccg ggaaatctcc
     7441 gccccgttcg taagccattt ccgctcgccg cagtcgaacg accgagcgta gcgagtcagt
     7501 gagcgaggaa gcggaatata tcccgaagcg gcatgcattt acgttgacac catcgaatgg
     7561 tgcaaaacct ttcgcggtat ggcatgatag cgcccggaag agagtcaatt cagggtggtg
     7621 aatgtgaaac cagtaacgtt atacgatgtc gcagagtatg ccggtgtctc ttatcagacc
     7681 gtttcccgcg tggtgaacca ggccagccac gtttctgcga aaacgcggga aaaagtggaa
     7741 gcggcgatgg cggagctgaa ttacattccc aaccgcgtgg cacaacaact ggcgggcaaa
     7801 cagtcgttgc tgattggcgt tgccacctcc agtctggccc tgcacgcgcc gtcgcaaatt
     7861 gtcgcggcga ttaaatctcg cgccgatcaa ctgggtgcca gcgtggtggt gtcgatggta
     7921 gaacgaagcg gcgtcgaagc ctgtaaagcg gcggtgcaca atcttctcgc gcaacgcgtc
     7981 agtgggctga tcattaacta tccgctggat gaccaggatg ccattgctgt ggaagctgcc
     8041 tgcactaatg ttccggcgtt atttcttgat gtctctgacc agacacccat caacagtatt
     8101 attttctccc atgaagacgg tacgcgactg ggcgtggagc atctggtcgc attgggtcac
     8161 cagcaaatcg cgctgttagc gggcccatta agttctgtct cggcgcgtct gcgtctggct
     8221 ggctggcata aatatctcac tcgcaatcaa attcagccga tagcggaacg ggaaggcgac
     8281 tggagtgcca tgtccggttt tcaacaaacc atgcaaatgc tgaatgaggg catcgttccc
     8341 actgcgatgc tggttgccaa cgatcagatg gcgctgggcg caatgcgcgc cattaccgag
     8401 tccgggctgc gcgttggtgc ggatatctcg gtagtgggat acgacgatac cgaagacagc
     8461 tcatgttata tcccgccgtt aaccaccatc aaacaggatt ttcgcctgct ggggcaaacc
     8521 agcgtggacc gcttgctgca actctctcag ggccaggcgg tgaagggcaa tcagctgttg
     8581 cccgtctcac tggtgaaaag aaaaaccacc ctggcgccca atacgcaaac cgcctctccc
     8641 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
     8701 cagtgaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct
     8761 gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc gccgccctag acctagggcg
     8821 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacg
//
