LOCUS       sequence-251162-        4839 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of SO_4427 under Ptac.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4839)
  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 4839)
  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: pKD282
FEATURES             Location/Qualifiers
     source          1..4839
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      812..898
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     promoter        join(1071..1076,1077..1092,1093..1099)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1107..1123
                     /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      2442..2513
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2529..2556
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             2756..3547
                     /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      3559..3653
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      3767..4312
                     /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     4213..4232
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4429..4446
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4485..4562
                     /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(4563..4839,1..806)
                     /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(4582..4601)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 gcgtggtggt gtcgatggta gaacgaagcg gcgtcgaagc ctgtaaagcg gcggtgcaca
       61 atcttctcgc gcaacgcgtc agtgggctga tcattaacta tccgctggat gaccaggatg
      121 ccattgctgt ggaagctgcc tgcactaatg ttccggcgtt atttcttgat gtctctgacc
      181 agacacccat caacagtatt attttctccc atgaagacgg tacgcgactg ggcgtggagc
      241 atctggtcgc attgggtcac cagcaaatcg cgctgttagc gggcccatta agttctgtct
      301 cggcgcgtct gcgtctggct ggctggcata aatatctcac tcgcaatcaa attcagccga
      361 tagcggaacg ggaaggcgac tggagtgcca tgtccggttt tcaacaaacc atgcaaatgc
      421 tgaatgaggg catcgttccc actgcgatgc tggttgccaa cgatcagatg gcgctgggcg
      481 caatgcgcgc cattaccgag tccgggctgc gcgttggtgc ggatatctcg gtagtgggat
      541 acgacgatac cgaagacagc tcatgttata tcccgccgtt aaccaccatc aaacaggatt
      601 ttcgcctgct ggggcaaacc agcgtggacc gcttgctgca actctctcag ggccaggcgg
      661 tgaagggcaa tcagctgttg cccgtctcac tggtgaaaag aaaaaccacc ctggcgccca
      721 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
      781 tttcccgact ggaaagcggg cagtgaggca tcaaataaaa cgaaaggctc agtcgaaaga
      841 ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc
      901 gccgccctag acctagggcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
      961 aatacgtaaa tcactgcata attcgtgtcg ctcaaggcgc actcccgttc tggataatgt
     1021 tttttgcgcc gacatcataa cggttctggc taatattctg aaatgagctg ttgacaatta
     1081 atcatcggct cgtataatgt gtggaattgt gagcggataa caatttcaca caaccctatt
     1141 ttattcttac ttcccttaga gaaagaaatg actgtgatca taggcgcttt gctgtttgct
     1201 ctctatcggc aattgattat cgagcagcag tatcaagtca ctcaatatct tgagtcagaa
     1261 actcagcgtt atcagcagct tgcgcttaca gtcgatcgcc gcagttttgc ggcacaaatt
     1321 cgtgccgccg atccgcaaac cgcgcttatc gcttggcgca atagttacga tatggtcggt
     1381 gctttaagct ttatgcccga aggcatgccc atgctgccac aaacgcggga ttttccgata
     1441 ttaaccggtg ggccagataa gctgcatatt cttaccggtg gtctggtaat gacccgctat
     1501 ggccccgtct tgattgccac tcgtaccgat aacttagcta ctcttatcga taaatttatc
     1561 agcgcggcag ccactgccgt gatgttgact atcgtgctga ccttggcgtt gggctatttg
     1621 ttttctaagg caattttacg ccgtttagtg caatacaacc gtttaagtga gcaaattgag
     1681 cgcggacatt atgacacgcg tttgccactc agttggcgtc aggacgagtt cgatatgtta
     1741 gcgctgcaat ttaataacgt gctggatatt ctcgaaaata acctgatggc ggtacgtggt
     1801 gtgacagata atatcgccca cgatttacgc acacccttat cccatatccg gattggtatt
     1861 gaggagttgg cgaccaagtc ccccgatgaa attagcgaag gttgtgcgat tttgaccgaa
     1921 gaactcgatc actgtcttgc gacttttgat gccatgctct ccctcacacg gattgaggag
     1981 ggtcagcaaa ccttagattt acaagagctg agtttggccc agctctgcac ggatttgctg
     2041 gaaatggccg atgcggtagc tgagtcgaat gagcaaacct tgagtctgtc gctacttgct
     2101 gatcataaag ttcatggcga taaatatttg ttattccaag ccttatataa tttggttgac
     2161 aatgctatga agtactcagg ccaaggcgct cgtatcgata ttattcagtc cggcgctcaa
     2221 attaagattt gcgataatgg cccagggatc cccgatgaca gcaaagaaag agtgtttgag
     2281 cgtttagttc gcctcgaccc aagccgccat ttgcaaggca caggtttagg attatctatg
     2341 gtcaaagcca ttctatcgcg gcacaatgcg cagattactc tttcggataa ccatcctggc
     2401 cttgtggtca ctatccagtt ttaataatct agaccaggca tcaaataaaa cgaaaggctc
     2461 agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctctactaga
     2521 gtcacactgg ctcaccttcg ggtgggcctt tctgcgttta taggtacccg gggatcctct
     2581 agagtcgacc tgcaggcatg caagcttagc aagcgaaccg gaattgccag ctggggcgcc
     2641 ctctggtaag gttgggaagc cctgcaaagt aaactggatg gctttcttgc cgccaaggat
     2701 ctgatggcgc aggggatcaa gatctgatca agagacagga tgaggatcgt ttcgtatgag
     2761 ggaagcggtg atcgccgaag tatcgactca actatcagag gtagttggcg tcatcgagcg
     2821 ccatctcgaa ccgacgttgc tggccgtaca tttgtacggc tccgcagtgg atggcggcct
     2881 gaagccacac agtgatattg atttgctggt tacggtgacc gtaaggcttg atgaaacaac
     2941 gcggcgagct ttgatcaacg accttttgga aacttcggct tcccctggag agagcgagat
     3001 tctccgcgct gtagaagtca ccattgttgt gcacgacgac atcattccgt ggcgttatcc
     3061 agctaagcgc gaactgcaat ttggagaatg gcagcgcaat gacattcttg caggtatctt
     3121 cgagccagcc acgatcgaca ttgatctggc tatcttgctg acaaaagcaa gagaacatag
     3181 cgttgccttg gtaggtccag cggcggagga actctttgat ccggttcctg aacaggatct
     3241 atttgaggcg ctaaatgaaa ccttaacgct atggaactcg ccgcccgact gggctggcga
     3301 tgagcgaaat gtagtgctta cgttgtcccg catttggtac agcgcagtaa ccggcaaaat
     3361 cgcgccgaag gatgtcgctg ccgactgggc aatggagcgc ctgccggccc agtatcagcc
     3421 cgtcatactt gaagctagac aggcttatct tggacaagaa gaagatcgct tggcctcgcg
     3481 cgcagatcag ttggaagaat ttgtccacta cgtgaaaggc gagatcacca aggtagtcgg
     3541 caaataagag ctcgcttgga ctcctgttga tagatccagt aatgacctca gaactccatc
     3601 tggatttgtt cagaacgctc ggttgccgcc gggcgttttt tattggtgag aatccaagca
     3661 ctagtaacaa cttatatcgt atggggctga cttcaggtgc tacatttgaa gagataaatt
     3721 gcactgaaat ctagtaatat tttatctgat taataagatg atcttcttga gatcgttttg
     3781 gtctgcgcgt aatctcttgc tctgaaaacg aaaaaaccgc cttgcagggc ggtttttcga
     3841 aggttctctg agctaccaac tctttgaacc gaggtaactg gcttggagga gcgcagtcac
     3901 caaaacttgt cctttcagtt tagccttaac cggcgcatga cttcaagact aactcctcta
     3961 aatcaattac cagtggctgc tgccagtggt gcttttgcat gtctttccgg gttggactca
     4021 agacgatagt taccggataa ggcgcagcgg tcggactgaa cggggggttc gtgcatacag
     4081 tccagcttgg agcgaactgc ctacccggaa ctgagtgtca ggcgtggaat gagacaaacg
     4141 cggccataac agcggaatga caccggtaaa ccgaaaggca ggaacaggag agcgcacgag
     4201 ggagccgcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccaccactg
     4261 atttgagcgt cagatttcgt gatgcttgtc aggggggcgg agcctatgga aaaacggctt
     4321 tgccgcggcc ctctcacttc cctgttaagt atcttcctgg catcttccgg gaaatctccg
     4381 ccccgttcgt aagccatttc cgctcgccgc agtcgaacga ccgagcgtag cgagtcagtg
     4441 agcgaggaag cggaatatat cccgaagcgg catgcattta cgttgacacc atcgaatggt
     4501 gcaaaacctt tcgcggtatg gcatgatagc gcccggaaga gagtcaattc agggtggtga
     4561 atgtgaaacc agtaacgtta tacgatgtcg cagagtatgc cggtgtctct tatcagaccg
     4621 tttcccgcgt ggtgaaccag gccagccacg tttctgcgaa aacgcgggaa aaagtggaag
     4681 cggcgatggc ggagctgaat tacattccca accgcgtggc acaacaactg gcgggcaaac
     4741 agtcgttgct gattggcgtt gccacctcca gtctggccct gcacgcgccg tcgcaaattg
     4801 tcgcggcgat taaatctcgc gccgatcaac tgggtgcca
//
