LOCUS       sequence-247909-        4899 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of SO_4488 under Ptac.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4899)
  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 4899)
  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: pKD284
FEATURES             Location/Qualifiers
     source          1..4899
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(76..81,82..97,98..104)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    112..128
                     /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      1507..1578
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1594..1621
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             1821..2612
                     /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      2624..2718
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      2832..3377
                     /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     3278..3297
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3494..3511
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        3550..3627
                     /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             3628..4710
                     /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(3647..3666)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     terminator      4716..4802
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ggcgcactcc cgttctggat aatgtttttt gcgccgacat cataacggtt ctggctaata
       61 ttctgaaatg agctgttgac aattaatcat cggctcgtat aatgtgtgga attgtgagcg
      121 gataacaatt tcacacatac cattcctata tagttcaact tattagttcg gtatgtattc
      181 acttaaggct tatctcactc gcaacctgct tatcaccctc acctgcgcca tggcgttatt
      241 gttgtatttg ctgtttcaag gcattcaaat cttaacccaa gactttgttg cctcgcgttt
      301 gcagcacgat accgatagtc tgatttccgc cctcgagccg caagtcgatg gcacttggtc
      361 actcaatgcc gatcgcctac ctaatgtgta tcacagggta aactcgggtc actactttgc
      421 aattcaaatc ggtgagcaac aactacgctc ccgctcactg tttgatcatg ccgttaccct
      481 gccaagcctg aaggtcgggg aacagcagca agccacccaa tatgttggcc aagaacattg
      541 gctgatgctc agccagacaa tcgtcaaaaa tgggcaatct attcgccttt gggtgacaga
      601 agacatcagc tcacttgagc aaacccagtt acgcttttta ggttttgcga ccacaatggt
      661 gctgctaacg atattgatct tactttttgc ccagcaccaa ctcctcaaac ggggttttaa
      721 accactcgaa aagatacctg aagccatcag acaaatgcgg ctccaaggaa gagagattga
      781 tactcagcag acgcccagcg agataacccc tttgattgct gaaatcgatc gcctagtaaa
      841 ccagctcggc caaagggttc aacgctcacg caatgccctt ggtaatcttg cccacgaaat
      901 gaaacgccct ctgcaagggc tgcaatccta cctcgatagc caagcgcctg aacaaagaca
      961 cgaaggcagt aaagtgctgg ccgaactgca ccatattatt gaacgagagc ttaagcgaac
     1021 caaaatcgtc gggctttcca caccgggacg ttttaccgtg cttaatgacg atctatcgcc
     1081 gctgatccaa gtcatgcaac gcatttatcc ccacaggcgg attagctgcc actatgacaa
     1141 agatctcgtc atgccctttg atagagatga cttactggaa ctcctcggca atttgctgga
     1201 taacgcctgc aagcatggcc acaaacacat cgaactctgc attatgcaac aatcagtacc
     1261 aaacccaggt tatgggatca tagcaagtga tgacggcgaa ggcgtctctg atgccgcaat
     1321 gtccatcata gttgagcgcg gtattaggct agacgaaagc attcaagggc atggcttagg
     1381 gctttcaatc tgcaaagaca ttgtcgacag ctatcagggg gagcttcact ttaaccatgc
     1441 caaacaaggt ggactcgaag ccaaggtgtt tctgccactt ccaaattaat aatctagacc
     1501 aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt
     1561 ttgtcggtga acgctctcta ctagagtcac actggctcac cttcgggtgg gcctttctgc
     1621 gtttataggt acccggggat cctctagagt cgacctgcag gcatgcaagc ttagcaagcg
     1681 aaccggaatt gccagctggg gcgccctctg gtaaggttgg gaagccctgc aaagtaaact
     1741 ggatggcttt cttgccgcca aggatctgat ggcgcagggg atcaagatct gatcaagaga
     1801 caggatgagg atcgtttcgt atgagggaag cggtgatcgc cgaagtatcg actcaactat
     1861 cagaggtagt tggcgtcatc gagcgccatc tcgaaccgac gttgctggcc gtacatttgt
     1921 acggctccgc agtggatggc ggcctgaagc cacacagtga tattgatttg ctggttacgg
     1981 tgaccgtaag gcttgatgaa acaacgcggc gagctttgat caacgacctt ttggaaactt
     2041 cggcttcccc tggagagagc gagattctcc gcgctgtaga agtcaccatt gttgtgcacg
     2101 acgacatcat tccgtggcgt tatccagcta agcgcgaact gcaatttgga gaatggcagc
     2161 gcaatgacat tcttgcaggt atcttcgagc cagccacgat cgacattgat ctggctatct
     2221 tgctgacaaa agcaagagaa catagcgttg ccttggtagg tccagcggcg gaggaactct
     2281 ttgatccggt tcctgaacag gatctatttg aggcgctaaa tgaaacctta acgctatgga
     2341 actcgccgcc cgactgggct ggcgatgagc gaaatgtagt gcttacgttg tcccgcattt
     2401 ggtacagcgc agtaaccggc aaaatcgcgc cgaaggatgt cgctgccgac tgggcaatgg
     2461 agcgcctgcc ggcccagtat cagcccgtca tacttgaagc tagacaggct tatcttggac
     2521 aagaagaaga tcgcttggcc tcgcgcgcag atcagttgga agaatttgtc cactacgtga
     2581 aaggcgagat caccaaggta gtcggcaaat aagagctcgc ttggactcct gttgatagat
     2641 ccagtaatga cctcagaact ccatctggat ttgttcagaa cgctcggttg ccgccgggcg
     2701 ttttttattg gtgagaatcc aagcactagt aacaacttat atcgtatggg gctgacttca
     2761 ggtgctacat ttgaagagat aaattgcact gaaatctagt aatattttat ctgattaata
     2821 agatgatctt cttgagatcg ttttggtctg cgcgtaatct cttgctctga aaacgaaaaa
     2881 accgccttgc agggcggttt ttcgaaggtt ctctgagcta ccaactcttt gaaccgaggt
     2941 aactggcttg gaggagcgca gtcaccaaaa cttgtccttt cagtttagcc ttaaccggcg
     3001 catgacttca agactaactc ctctaaatca attaccagtg gctgctgcca gtggtgcttt
     3061 tgcatgtctt tccgggttgg actcaagacg atagttaccg gataaggcgc agcggtcgga
     3121 ctgaacgggg ggttcgtgca tacagtccag cttggagcga actgcctacc cggaactgag
     3181 tgtcaggcgt ggaatgagac aaacgcggcc ataacagcgg aatgacaccg gtaaaccgaa
     3241 aggcaggaac aggagagcgc acgagggagc cgccaggggg aaacgcctgg tatctttata
     3301 gtcctgtcgg gtttcgccac cactgatttg agcgtcagat ttcgtgatgc ttgtcagggg
     3361 ggcggagcct atggaaaaac ggctttgccg cggccctctc acttccctgt taagtatctt
     3421 cctggcatct tccgggaaat ctccgccccg ttcgtaagcc atttccgctc gccgcagtcg
     3481 aacgaccgag cgtagcgagt cagtgagcga ggaagcggaa tatatcccga agcggcatgc
     3541 atttacgttg acaccatcga atggtgcaaa acctttcgcg gtatggcatg atagcgcccg
     3601 gaagagagtc aattcagggt ggtgaatgtg aaaccagtaa cgttatacga tgtcgcagag
     3661 tatgccggtg tctcttatca gaccgtttcc cgcgtggtga accaggccag ccacgtttct
     3721 gcgaaaacgc gggaaaaagt ggaagcggcg atggcggagc tgaattacat tcccaaccgc
     3781 gtggcacaac aactggcggg caaacagtcg ttgctgattg gcgttgccac ctccagtctg
     3841 gccctgcacg cgccgtcgca aattgtcgcg gcgattaaat ctcgcgccga tcaactgggt
     3901 gccagcgtgg tggtgtcgat ggtagaacga agcggcgtcg aagcctgtaa agcggcggtg
     3961 cacaatcttc tcgcgcaacg cgtcagtggg ctgatcatta actatccgct ggatgaccag
     4021 gatgccattg ctgtggaagc tgcctgcact aatgttccgg cgttatttct tgatgtctct
     4081 gaccagacac ccatcaacag tattattttc tcccatgaag acggtacgcg actgggcgtg
     4141 gagcatctgg tcgcattggg tcaccagcaa atcgcgctgt tagcgggccc attaagttct
     4201 gtctcggcgc gtctgcgtct ggctggctgg cataaatatc tcactcgcaa tcaaattcag
     4261 ccgatagcgg aacgggaagg cgactggagt gccatgtccg gttttcaaca aaccatgcaa
     4321 atgctgaatg agggcatcgt tcccactgcg atgctggttg ccaacgatca gatggcgctg
     4381 ggcgcaatgc gcgccattac cgagtccggg ctgcgcgttg gtgcggatat ctcggtagtg
     4441 ggatacgacg ataccgaaga cagctcatgt tatatcccgc cgttaaccac catcaaacag
     4501 gattttcgcc tgctggggca aaccagcgtg gaccgcttgc tgcaactctc tcagggccag
     4561 gcggtgaagg gcaatcagct gttgcccgtc tcactggtga aaagaaaaac caccctggcg
     4621 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
     4681 caggtttccc gactggaaag cgggcagtga ggcatcaaat aaaacgaaag gctcagtcga
     4741 aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa
     4801 atccgccgcc ctagacctag ggcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     4861 cggtaatacg taaatcactg cataattcgt gtcgctcaa
//
