LOCUS       sequence-251164-        4873 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of SO_4638 under Ptac.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4873)
  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 4873)
  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: pKD288
FEATURES             Location/Qualifiers
     source          1..4873
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(84..89,90..105,106..112)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    120..136
                     /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      1489..1560
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1576..1603
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             1803..2594
                     /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      2606..2700
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      2814..3359
                     /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     3260..3279
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3476..3493
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        3532..3609
                     /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             3610..4692
                     /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(3629..3648)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     terminator      4698..4784
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 tcgctcaagg cgcactcccg ttctggataa tgttttttgc gccgacatca taacggttct
       61 ggctaatatt ctgaaatgag ctgttgacaa ttaatcatcg gctcgtataa tgtgtggaat
      121 tgtgagcgga taacaatttc acacacgaat agataaacct tcgtaaagga cacaataaat
      181 ggccaaggtt accgactcct cccctaagcc attaccgagc ttatacaaaa aaattcgcct
      241 aacgttttta gcgctcacaa gcctaatgtt cgcgattttc tggggtttga tttatatcgc
      301 cgaagaacaa attgaggtca ttagccttga acacaaactg gcgaccgaaa cccgtgaata
      361 ccagaaaaac tatcaacgat ttggcgataa agcccagctc cccgatatcc acgaactcac
      421 cacctattgg agcaaagagt ttgtgccgca atggctgcaa gcctatcaac aaacgggttt
      481 ttatgaacac cagttgggtg ccgaagataa gcactttttg atttcgccgc acccatcggg
      541 gcaaggttta ttgtatgtgg tgtttaagga tgatgcggat gactttctcg atggatatga
      601 ggacaggctg catattttga tatccctctt cggggtaggt ttactgctgt tcatcatggc
      661 ctatggctta tattttgtgc ggctactatc ggcgccattt tttgaaattg aacgaaaaat
      721 caatgcgctg acgccagagg agccgagctt tatccctagc acgccctacc gtgaaacccg
      781 ccatatcgaa caaaccttac tggaaaataa aaccaaaatt gccgcctatt ttactcggga
      841 acaggagttt agccgctttg cctcccatga actacgcacg cctattatgg tgatccaagg
      901 ctcgacagaa ttactcaaaa aactgccaaa cagcgcgaac gaagcccagc agcggatcgc
      961 caacaaagcc atcgcccgca ttgattacgc aggccaacag atgaaaaccc tcaccgaaac
     1021 ctttttgtta ttaggtaagg aacaaattga atcgcaacac tatgatcaag ttgatttagc
     1081 cgcacggctt acgcagctat taagtgaaat ggcgcagcat tttgccaaac acgaaatccg
     1141 ccatcagctc agcgttaacc aaaataccgc catttgcgcg ccggtgagtt ttgtggacat
     1201 agtgataggc aacttaatta aaaatgcctt taactacagc agtggcgata ttgaaatcac
     1261 gctgaaaaac aatcaactaa ccataaaaaa tccgcttagc caccacagcg ccgagcccac
     1321 gggctatggt tgtggtttga ttattattga acgtatctgc gagcgcatgg ggtggcactt
     1381 ttcggcccaa agcctagggg atgattatct ggcaacaatc caatttatgg ataaccgcga
     1441 gccacaacag catctaaatg aatccgctta ataatctaga ccaggcatca aataaaacga
     1501 aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc
     1561 tactagagtc acactggctc accttcgggt gggcctttct gcgtttatag gtacccgggg
     1621 atcctctaga gtcgacctgc aggcatgcaa gcttagcaag cgaaccggaa ttgccagctg
     1681 gggcgccctc tggtaaggtt gggaagccct gcaaagtaaa ctggatggct ttcttgccgc
     1741 caaggatctg atggcgcagg ggatcaagat ctgatcaaga gacaggatga ggatcgtttc
     1801 gtatgaggga agcggtgatc gccgaagtat cgactcaact atcagaggta gttggcgtca
     1861 tcgagcgcca tctcgaaccg acgttgctgg ccgtacattt gtacggctcc gcagtggatg
     1921 gcggcctgaa gccacacagt gatattgatt tgctggttac ggtgaccgta aggcttgatg
     1981 aaacaacgcg gcgagctttg atcaacgacc ttttggaaac ttcggcttcc cctggagaga
     2041 gcgagattct ccgcgctgta gaagtcacca ttgttgtgca cgacgacatc attccgtggc
     2101 gttatccagc taagcgcgaa ctgcaatttg gagaatggca gcgcaatgac attcttgcag
     2161 gtatcttcga gccagccacg atcgacattg atctggctat cttgctgaca aaagcaagag
     2221 aacatagcgt tgccttggta ggtccagcgg cggaggaact ctttgatccg gttcctgaac
     2281 aggatctatt tgaggcgcta aatgaaacct taacgctatg gaactcgccg cccgactggg
     2341 ctggcgatga gcgaaatgta gtgcttacgt tgtcccgcat ttggtacagc gcagtaaccg
     2401 gcaaaatcgc gccgaaggat gtcgctgccg actgggcaat ggagcgcctg ccggcccagt
     2461 atcagcccgt catacttgaa gctagacagg cttatcttgg acaagaagaa gatcgcttgg
     2521 cctcgcgcgc agatcagttg gaagaatttg tccactacgt gaaaggcgag atcaccaagg
     2581 tagtcggcaa ataagagctc gcttggactc ctgttgatag atccagtaat gacctcagaa
     2641 ctccatctgg atttgttcag aacgctcggt tgccgccggg cgttttttat tggtgagaat
     2701 ccaagcacta gtaacaactt atatcgtatg gggctgactt caggtgctac atttgaagag
     2761 ataaattgca ctgaaatcta gtaatatttt atctgattaa taagatgatc ttcttgagat
     2821 cgttttggtc tgcgcgtaat ctcttgctct gaaaacgaaa aaaccgcctt gcagggcggt
     2881 ttttcgaagg ttctctgagc taccaactct ttgaaccgag gtaactggct tggaggagcg
     2941 cagtcaccaa aacttgtcct ttcagtttag ccttaaccgg cgcatgactt caagactaac
     3001 tcctctaaat caattaccag tggctgctgc cagtggtgct tttgcatgtc tttccgggtt
     3061 ggactcaaga cgatagttac cggataaggc gcagcggtcg gactgaacgg ggggttcgtg
     3121 catacagtcc agcttggagc gaactgccta cccggaactg agtgtcaggc gtggaatgag
     3181 acaaacgcgg ccataacagc ggaatgacac cggtaaaccg aaaggcagga acaggagagc
     3241 gcacgaggga gccgccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     3301 accactgatt tgagcgtcag atttcgtgat gcttgtcagg ggggcggagc ctatggaaaa
     3361 acggctttgc cgcggccctc tcacttccct gttaagtatc ttcctggcat cttccgggaa
     3421 atctccgccc cgttcgtaag ccatttccgc tcgccgcagt cgaacgaccg agcgtagcga
     3481 gtcagtgagc gaggaagcgg aatatatccc gaagcggcat gcatttacgt tgacaccatc
     3541 gaatggtgca aaacctttcg cggtatggca tgatagcgcc cggaagagag tcaattcagg
     3601 gtggtgaatg tgaaaccagt aacgttatac gatgtcgcag agtatgccgg tgtctcttat
     3661 cagaccgttt cccgcgtggt gaaccaggcc agccacgttt ctgcgaaaac gcgggaaaaa
     3721 gtggaagcgg cgatggcgga gctgaattac attcccaacc gcgtggcaca acaactggcg
     3781 ggcaaacagt cgttgctgat tggcgttgcc acctccagtc tggccctgca cgcgccgtcg
     3841 caaattgtcg cggcgattaa atctcgcgcc gatcaactgg gtgccagcgt ggtggtgtcg
     3901 atggtagaac gaagcggcgt cgaagcctgt aaagcggcgg tgcacaatct tctcgcgcaa
     3961 cgcgtcagtg ggctgatcat taactatccg ctggatgacc aggatgccat tgctgtggaa
     4021 gctgcctgca ctaatgttcc ggcgttattt cttgatgtct ctgaccagac acccatcaac
     4081 agtattattt tctcccatga agacggtacg cgactgggcg tggagcatct ggtcgcattg
     4141 ggtcaccagc aaatcgcgct gttagcgggc ccattaagtt ctgtctcggc gcgtctgcgt
     4201 ctggctggct ggcataaata tctcactcgc aatcaaattc agccgatagc ggaacgggaa
     4261 ggcgactgga gtgccatgtc cggttttcaa caaaccatgc aaatgctgaa tgagggcatc
     4321 gttcccactg cgatgctggt tgccaacgat cagatggcgc tgggcgcaat gcgcgccatt
     4381 accgagtccg ggctgcgcgt tggtgcggat atctcggtag tgggatacga cgataccgaa
     4441 gacagctcat gttatatccc gccgttaacc accatcaaac aggattttcg cctgctgggg
     4501 caaaccagcg tggaccgctt gctgcaactc tctcagggcc aggcggtgaa gggcaatcag
     4561 ctgttgcccg tctcactggt gaaaagaaaa accaccctgg cgcccaatac gcaaaccgcc
     4621 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
     4681 agcgggcagt gaggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt
     4741 ttatctgttg tttgtcggtg aacgctctcc tgagtaggac aaatccgccg ccctagacct
     4801 agggcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cgtaaatcac
     4861 tgcataattc gtg
//
