LOCUS       sequence-247906-        4815 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of SO_4622 under Ptac.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4815)
  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 4815)
  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: pKD286
FEATURES             Location/Qualifiers
     source          1..4815
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(62..67,68..83,84..90)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    98..114
                     /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)."
     primer_bind     103..125
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     regulatory      1267..1276
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     terminator      1409..1480
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1496..1523
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             1723..2514
                     /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      2526..2620
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      2734..3279
                     /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     3180..3199
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3396..3413
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        3452..3529
                     /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             3530..4612
                     /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(3549..3568)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     terminator      4618..4704
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ctggataatg ttttttgcgc cgacatcata acggttctgg ctaatattct gaaatgagct
       61 gttgacaatt aatcatcggc tcgtataatg tgtggaattg tgagcggata acaatttcac
      121 acaggacacc actcactctt tatccttcaa tgcaagcgcc taaatttagc ctcaagcgta
      181 gctatcaagt gtggggagtt ttttttcttt taataacctt aggtgtgaat agttttatcc
      241 cgctttgcat gatgtgtgca ggtatgattt ctatgcatga agtcacaatg cataatgcag
      301 ggcaagcggc cgaagcatta cttaaggtgg cacagccaac tcagcaacca ttacagcagc
      361 cgattctaaa aactgacaac ttaacagtct atcctcattg gctagatatt ccgccgcaca
      421 ttcgtgagtt aagtaacaat gaagtcttaa cggatgcgag caaaattcgc tatgagcatg
      481 cagaaggtcc cattgttaac gcgctgagtg tgtactacac ccaagctggc cagccgctct
      541 acgtttggct gcagtttagt gttacccatg atttgcactt tgcccctaaa aatatgacgg
      601 ccattttgct gcttgtcttg ataacctttt gtgccagtgc aagcttagcg ttttatatgc
      661 aaagcaaagt gattacgcct gtgcatcaaa ttagccaagc gattaaacag catgattggg
      721 aggcaggagg acaactgagt ttgccgcaac aacagtatgc agaacttaaa gccattgttg
      781 atgctttaga ttactcgatt ttacagcttc aagaggctca gcaacgcgaa cttaattttt
      841 tacgctttgc tagccatgag ttaagaacgc cagttgctat atgccaatct tcattcgaaa
      901 tcctcgcatt acaacaaggt gagcttacgg gggcaacgct gtatgcttcc caagcgacca
      961 atcaaatgaa gtcactcatt gaaaccttac tgtggttgac taataaagag tgttccagct
     1021 tagaaaagga aaatgtcgcg ttatcgcctc tacttgcctg tattatcgag gagcagcaac
     1081 gggtacatgg tcatgagcaa aaaattcaat tattttctga cgatacttgt ttgcccattc
     1141 agcttcatcc gctaaaaata atattaagca atttattaag gaatagtatt gagcatggag
     1201 ggcgggatat tcgcattact caagatgctt gttgtataga aataaccaat tcctacaata
     1261 gtaagggccg ccatggattt ggtttagggc ttttactggt gaaacgtctc gcaaaccaat
     1321 tgggttggtt ttttactagc tcatctgaaa ataatttatt taccgctagg ttggaaatat
     1381 ctaacagtta ataatctaga ccaggcatca aataaaacga aaggctcagt cgaaagactg
     1441 ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc tactagagtc acactggctc
     1501 accttcgggt gggcctttct gcgtttatag gtacccgggg atcctctaga gtcgacctgc
     1561 aggcatgcaa gcttagcaag cgaaccggaa ttgccagctg gggcgccctc tggtaaggtt
     1621 gggaagccct gcaaagtaaa ctggatggct ttcttgccgc caaggatctg atggcgcagg
     1681 ggatcaagat ctgatcaaga gacaggatga ggatcgtttc gtatgaggga agcggtgatc
     1741 gccgaagtat cgactcaact atcagaggta gttggcgtca tcgagcgcca tctcgaaccg
     1801 acgttgctgg ccgtacattt gtacggctcc gcagtggatg gcggcctgaa gccacacagt
     1861 gatattgatt tgctggttac ggtgaccgta aggcttgatg aaacaacgcg gcgagctttg
     1921 atcaacgacc ttttggaaac ttcggcttcc cctggagaga gcgagattct ccgcgctgta
     1981 gaagtcacca ttgttgtgca cgacgacatc attccgtggc gttatccagc taagcgcgaa
     2041 ctgcaatttg gagaatggca gcgcaatgac attcttgcag gtatcttcga gccagccacg
     2101 atcgacattg atctggctat cttgctgaca aaagcaagag aacatagcgt tgccttggta
     2161 ggtccagcgg cggaggaact ctttgatccg gttcctgaac aggatctatt tgaggcgcta
     2221 aatgaaacct taacgctatg gaactcgccg cccgactggg ctggcgatga gcgaaatgta
     2281 gtgcttacgt tgtcccgcat ttggtacagc gcagtaaccg gcaaaatcgc gccgaaggat
     2341 gtcgctgccg actgggcaat ggagcgcctg ccggcccagt atcagcccgt catacttgaa
     2401 gctagacagg cttatcttgg acaagaagaa gatcgcttgg cctcgcgcgc agatcagttg
     2461 gaagaatttg tccactacgt gaaaggcgag atcaccaagg tagtcggcaa ataagagctc
     2521 gcttggactc ctgttgatag atccagtaat gacctcagaa ctccatctgg atttgttcag
     2581 aacgctcggt tgccgccggg cgttttttat tggtgagaat ccaagcacta gtaacaactt
     2641 atatcgtatg gggctgactt caggtgctac atttgaagag ataaattgca ctgaaatcta
     2701 gtaatatttt atctgattaa taagatgatc ttcttgagat cgttttggtc tgcgcgtaat
     2761 ctcttgctct gaaaacgaaa aaaccgcctt gcagggcggt ttttcgaagg ttctctgagc
     2821 taccaactct ttgaaccgag gtaactggct tggaggagcg cagtcaccaa aacttgtcct
     2881 ttcagtttag ccttaaccgg cgcatgactt caagactaac tcctctaaat caattaccag
     2941 tggctgctgc cagtggtgct tttgcatgtc tttccgggtt ggactcaaga cgatagttac
     3001 cggataaggc gcagcggtcg gactgaacgg ggggttcgtg catacagtcc agcttggagc
     3061 gaactgccta cccggaactg agtgtcaggc gtggaatgag acaaacgcgg ccataacagc
     3121 ggaatgacac cggtaaaccg aaaggcagga acaggagagc gcacgaggga gccgccaggg
     3181 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc accactgatt tgagcgtcag
     3241 atttcgtgat gcttgtcagg ggggcggagc ctatggaaaa acggctttgc cgcggccctc
     3301 tcacttccct gttaagtatc ttcctggcat cttccgggaa atctccgccc cgttcgtaag
     3361 ccatttccgc tcgccgcagt cgaacgaccg agcgtagcga gtcagtgagc gaggaagcgg
     3421 aatatatccc gaagcggcat gcatttacgt tgacaccatc gaatggtgca aaacctttcg
     3481 cggtatggca tgatagcgcc cggaagagag tcaattcagg gtggtgaatg tgaaaccagt
     3541 aacgttatac gatgtcgcag agtatgccgg tgtctcttat cagaccgttt cccgcgtggt
     3601 gaaccaggcc agccacgttt ctgcgaaaac gcgggaaaaa gtggaagcgg cgatggcgga
     3661 gctgaattac attcccaacc gcgtggcaca acaactggcg ggcaaacagt cgttgctgat
     3721 tggcgttgcc acctccagtc tggccctgca cgcgccgtcg caaattgtcg cggcgattaa
     3781 atctcgcgcc gatcaactgg gtgccagcgt ggtggtgtcg atggtagaac gaagcggcgt
     3841 cgaagcctgt aaagcggcgg tgcacaatct tctcgcgcaa cgcgtcagtg ggctgatcat
     3901 taactatccg ctggatgacc aggatgccat tgctgtggaa gctgcctgca ctaatgttcc
     3961 ggcgttattt cttgatgtct ctgaccagac acccatcaac agtattattt tctcccatga
     4021 agacggtacg cgactgggcg tggagcatct ggtcgcattg ggtcaccagc aaatcgcgct
     4081 gttagcgggc ccattaagtt ctgtctcggc gcgtctgcgt ctggctggct ggcataaata
     4141 tctcactcgc aatcaaattc agccgatagc ggaacgggaa ggcgactgga gtgccatgtc
     4201 cggttttcaa caaaccatgc aaatgctgaa tgagggcatc gttcccactg cgatgctggt
     4261 tgccaacgat cagatggcgc tgggcgcaat gcgcgccatt accgagtccg ggctgcgcgt
     4321 tggtgcggat atctcggtag tgggatacga cgataccgaa gacagctcat gttatatccc
     4381 gccgttaacc accatcaaac aggattttcg cctgctgggg caaaccagcg tggaccgctt
     4441 gctgcaactc tctcagggcc aggcggtgaa gggcaatcag ctgttgcccg tctcactggt
     4501 gaaaagaaaa accaccctgg cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
     4561 ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt gaggcatcaa
     4621 ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg
     4681 aacgctctcc tgagtaggac aaatccgccg ccctagacct agggcgttcg gctgcggcga
     4741 gcggtatcag ctcactcaaa ggcggtaata cgtaaatcac tgcataattc gtgtcgctca
     4801 aggcgcactc ccgtt
//
