LOCUS       sequence-247910-        5742 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of SO_2192 under Ptac.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5742)
  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 5742)
  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: pKD278
FEATURES             Location/Qualifiers
     source          1..5742
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      668..754
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     promoter        join(927..932,933..948,949..955)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    963..979
                     /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      3201..3272
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3288..3315
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             3515..4306
                     /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      4318..4412
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      4526..5071
                     /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     4972..4991
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5188..5205
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        5244..5321
                     /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             join(5322..5742,1..662)
                     /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(5341..5360)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 ctaatgttcc ggcgttattt cttgatgtct ctgaccagac acccatcaac agtattattt
       61 tctcccatga agacggtacg cgactgggcg tggagcatct ggtcgcattg ggtcaccagc
      121 aaatcgcgct gttagcgggc ccattaagtt ctgtctcggc gcgtctgcgt ctggctggct
      181 ggcataaata tctcactcgc aatcaaattc agccgatagc ggaacgggaa ggcgactgga
      241 gtgccatgtc cggttttcaa caaaccatgc aaatgctgaa tgagggcatc gttcccactg
      301 cgatgctggt tgccaacgat cagatggcgc tgggcgcaat gcgcgccatt accgagtccg
      361 ggctgcgcgt tggtgcggat atctcggtag tgggatacga cgataccgaa gacagctcat
      421 gttatatccc gccgttaacc accatcaaac aggattttcg cctgctgggg caaaccagcg
      481 tggaccgctt gctgcaactc tctcagggcc aggcggtgaa gggcaatcag ctgttgcccg
      541 tctcactggt gaaaagaaaa accaccctgg cgcccaatac gcaaaccgcc tctccccgcg
      601 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
      661 gaggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg
      721 tttgtcggtg aacgctctcc tgagtaggac aaatccgccg ccctagacct agggcgttcg
      781 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cgtaaatcac tgcataattc
      841 gtgtcgctca aggcgcactc ccgttctgga taatgttttt tgcgccgaca tcataacggt
      901 tctggctaat attctgaaat gagctgttga caattaatca tcggctcgta taatgtgtgg
      961 aattgtgagc ggataacaat ttcacacaca ctattctcat ctaaatttat caaaacgcct
     1021 acaatgacac gttttccttt cggtcttcgc actaaagtga ttggcctgag cctatttttg
     1081 ctctgcctac cttggctggg gtaccaatat gtgtgggaga tggaaaaata cctgcgccac
     1141 ggccaagaac gtaccttaga gggcacaacc caagcgctag cgaccgccct gcacgagcgg
     1201 cccaaacttt tcgatggcca agccagtttt ttaaaacagg ttgaaaagag ccgtgacctc
     1261 tatgcttacc ccttagccgg agccattctg ctcgatggca aactcaatga atggcaggaa
     1321 tatcaacatc gatttatcca atacggtgaa gccaatcaga ttatgcgcgc cgatcccagt
     1381 gcgccgctca ccttaagttt tacccatatg gtgggccagt acgatggcta tctgtatgct
     1441 ttttttcagg tggtggaccc caatgtggtg tttcgtggca aaaatgccct aagtgtggat
     1501 aggaacgatc atctatcaat tgcgaccctt gcccccgaca ataaatttaa gcgttatatc
     1561 gttgccacca ctcagcccgg ttggatcagc gcctttgaac tgcccgccaa cccgaaacaa
     1621 accgtgccag tgacacccga ggtgcgcatt caaggacaat gggccaccac agatgtgggc
     1681 tacaatatcg agctgcgtat tccccttgat atggtgggca gcaaactcgg gtttgccatt
     1741 gccgatgtta acgacagcaa ataccgcgat gtcgccgcag tggttggcac ttccgcctta
     1801 gataccgtcg ataagctggg cacagtgctg gtcccctccc ctgaaattga aaacattatc
     1861 aaagggatga gccactacag ctcgcgtatt tgggtaatag ataaacatgg ccgcgtgtta
     1921 gcaaaatccg gcgatatccg cgacagcaac agcgtgtgga ctcgcaccct tgatgatgaa
     1981 aacagcaatg gcctatggca gcggtttcag cgcgattatc tgcatccgct ctactataaa
     2041 atcctcacta aaccgccgaa ggattttatc gactctctgc gggattcaac cgagctaaat
     2101 ggcagtcata ttcaaaaagc cctcgccgga caacagacgt cgacatggcg cttgacgcct
     2161 gacaataagg ccgtggtact cgcggcagcc agcccaattt ggattgataa caatgtgatg
     2221 ggcgcggtag tggctgaaga aaccacccac ggtattcgca ccctacgtaa tcgcgcgttg
     2281 gaaaaactgt ttaacgtgat cctgaccatt atgagcatgg gcaccttagc gctgtttttc
     2341 ttcgcatcga gtatttccaa tcgcattcgt aagttaaggg acgaagccga aaatgccatc
     2401 gacagccaag gtcgggtgcg taaccatcta aaaccctcaa aggcgcggga cgagattggc
     2461 gacttatccc gcagttttgc cagcattgta ggacgtttag gccaatacac tcattatttg
     2521 gagaatatgt cgtcgcgctt atcccatgag ctacgtactc ctgttgcagt ggtgcgttca
     2581 tctttagagc acttaaatct gcaaacctta gataaagaca cgcaaaagta tgtcgatcgc
     2641 gcccaagaag gcgtaagccg tttaagcatg attttgaata atatgagcga ggcgacgcgc
     2701 ctagagcaga gcttaactca ggccgatgtg actaagtttc cgctatccca agtggtcagc
     2761 ggctgtatgc aaggctatca aatgacctat cccgagcagc gttttagcat caaagtgccc
     2821 gatcagccgt tgatgatgca gggtgtgcca gaatacatcg cccaattgat ggataaactt
     2881 atcgccaatg ccatcgaatt ttgtcatcaa ggcagttcga ttgaggtcac cctcactcaa
     2941 gtcggtaaac aagccacatt gcttatcagc aacttagggc cgcagttgcc ggtggatatg
     3001 tcggagcaga tcttcgactc aatggtatcg gtgagggata atcaaaccca ggagaaaccc
     3061 caccttggcc taggtttgta tattgcacgg ctcgtgaccg agtttcacca agggaatatc
     3121 ttagcccgca accgcagcga aaatgatggg gtcgatatcc tagtcaccct gccgctgtgc
     3181 taataatcta gaccaggcat caaataaaac gaaaggctca gtcgaaagac tgggcctttc
     3241 gttttatctg ttgtttgtcg gtgaacgctc tctactagag tcacactggc tcaccttcgg
     3301 gtgggccttt ctgcgtttat aggtacccgg ggatcctcta gagtcgacct gcaggcatgc
     3361 aagcttagca agcgaaccgg aattgccagc tggggcgccc tctggtaagg ttgggaagcc
     3421 ctgcaaagta aactggatgg ctttcttgcc gccaaggatc tgatggcgca ggggatcaag
     3481 atctgatcaa gagacaggat gaggatcgtt tcgtatgagg gaagcggtga tcgccgaagt
     3541 atcgactcaa ctatcagagg tagttggcgt catcgagcgc catctcgaac cgacgttgct
     3601 ggccgtacat ttgtacggct ccgcagtgga tggcggcctg aagccacaca gtgatattga
     3661 tttgctggtt acggtgaccg taaggcttga tgaaacaacg cggcgagctt tgatcaacga
     3721 ccttttggaa acttcggctt cccctggaga gagcgagatt ctccgcgctg tagaagtcac
     3781 cattgttgtg cacgacgaca tcattccgtg gcgttatcca gctaagcgcg aactgcaatt
     3841 tggagaatgg cagcgcaatg acattcttgc aggtatcttc gagccagcca cgatcgacat
     3901 tgatctggct atcttgctga caaaagcaag agaacatagc gttgccttgg taggtccagc
     3961 ggcggaggaa ctctttgatc cggttcctga acaggatcta tttgaggcgc taaatgaaac
     4021 cttaacgcta tggaactcgc cgcccgactg ggctggcgat gagcgaaatg tagtgcttac
     4081 gttgtcccgc atttggtaca gcgcagtaac cggcaaaatc gcgccgaagg atgtcgctgc
     4141 cgactgggca atggagcgcc tgccggccca gtatcagccc gtcatacttg aagctagaca
     4201 ggcttatctt ggacaagaag aagatcgctt ggcctcgcgc gcagatcagt tggaagaatt
     4261 tgtccactac gtgaaaggcg agatcaccaa ggtagtcggc aaataagagc tcgcttggac
     4321 tcctgttgat agatccagta atgacctcag aactccatct ggatttgttc agaacgctcg
     4381 gttgccgccg ggcgtttttt attggtgaga atccaagcac tagtaacaac ttatatcgta
     4441 tggggctgac ttcaggtgct acatttgaag agataaattg cactgaaatc tagtaatatt
     4501 ttatctgatt aataagatga tcttcttgag atcgttttgg tctgcgcgta atctcttgct
     4561 ctgaaaacga aaaaaccgcc ttgcagggcg gtttttcgaa ggttctctga gctaccaact
     4621 ctttgaaccg aggtaactgg cttggaggag cgcagtcacc aaaacttgtc ctttcagttt
     4681 agccttaacc ggcgcatgac ttcaagacta actcctctaa atcaattacc agtggctgct
     4741 gccagtggtg cttttgcatg tctttccggg ttggactcaa gacgatagtt accggataag
     4801 gcgcagcggt cggactgaac ggggggttcg tgcatacagt ccagcttgga gcgaactgcc
     4861 tacccggaac tgagtgtcag gcgtggaatg agacaaacgc ggccataaca gcggaatgac
     4921 accggtaaac cgaaaggcag gaacaggaga gcgcacgagg gagccgccag ggggaaacgc
     4981 ctggtatctt tatagtcctg tcgggtttcg ccaccactga tttgagcgtc agatttcgtg
     5041 atgcttgtca ggggggcgga gcctatggaa aaacggcttt gccgcggccc tctcacttcc
     5101 ctgttaagta tcttcctggc atcttccggg aaatctccgc cccgttcgta agccatttcc
     5161 gctcgccgca gtcgaacgac cgagcgtagc gagtcagtga gcgaggaagc ggaatatatc
     5221 ccgaagcggc atgcatttac gttgacacca tcgaatggtg caaaaccttt cgcggtatgg
     5281 catgatagcg cccggaagag agtcaattca gggtggtgaa tgtgaaacca gtaacgttat
     5341 acgatgtcgc agagtatgcc ggtgtctctt atcagaccgt ttcccgcgtg gtgaaccagg
     5401 ccagccacgt ttctgcgaaa acgcgggaaa aagtggaagc ggcgatggcg gagctgaatt
     5461 acattcccaa ccgcgtggca caacaactgg cgggcaaaca gtcgttgctg attggcgttg
     5521 ccacctccag tctggccctg cacgcgccgt cgcaaattgt cgcggcgatt aaatctcgcg
     5581 ccgatcaact gggtgccagc gtggtggtgt cgatggtaga acgaagcggc gtcgaagcct
     5641 gtaaagcggc ggtgcacaat cttctcgcgc aacgcgtcag tgggctgatc attaactatc
     5701 cgctggatga ccaggatgcc attgctgtgg aagctgcctg ca
//
