LOCUS       sequence-247749-        4908 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of SO_0621 under Ptac.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4908)
  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 4908)
  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: pKD276
FEATURES             Location/Qualifiers
     source          1..4908
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     61..78
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        117..194
                     /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             195..1277
                     /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(214..233)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     terminator      1283..1369
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     promoter        join(1542..1547,1548..1563,1564..1570)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1578..1594
                     /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      2982..3053
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3069..3096
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             3296..4087
                     /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      4099..4193
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      4307..4852
                     /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     4753..4772
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gggaaatctc cgccccgttc gtaagccatt tccgctcgcc gcagtcgaac gaccgagcgt
       61 agcgagtcag tgagcgagga agcggaatat atcccgaagc ggcatgcatt tacgttgaca
      121 ccatcgaatg gtgcaaaacc tttcgcggta tggcatgata gcgcccggaa gagagtcaat
      181 tcagggtggt gaatgtgaaa ccagtaacgt tatacgatgt cgcagagtat gccggtgtct
      241 cttatcagac cgtttcccgc gtggtgaacc aggccagcca cgtttctgcg aaaacgcggg
      301 aaaaagtgga agcggcgatg gcggagctga attacattcc caaccgcgtg gcacaacaac
      361 tggcgggcaa acagtcgttg ctgattggcg ttgccacctc cagtctggcc ctgcacgcgc
      421 cgtcgcaaat tgtcgcggcg attaaatctc gcgccgatca actgggtgcc agcgtggtgg
      481 tgtcgatggt agaacgaagc ggcgtcgaag cctgtaaagc ggcggtgcac aatcttctcg
      541 cgcaacgcgt cagtgggctg atcattaact atccgctgga tgaccaggat gccattgctg
      601 tggaagctgc ctgcactaat gttccggcgt tatttcttga tgtctctgac cagacaccca
      661 tcaacagtat tattttctcc catgaagacg gtacgcgact gggcgtggag catctggtcg
      721 cattgggtca ccagcaaatc gcgctgttag cgggcccatt aagttctgtc tcggcgcgtc
      781 tgcgtctggc tggctggcat aaatatctca ctcgcaatca aattcagccg atagcggaac
      841 gggaaggcga ctggagtgcc atgtccggtt ttcaacaaac catgcaaatg ctgaatgagg
      901 gcatcgttcc cactgcgatg ctggttgcca acgatcagat ggcgctgggc gcaatgcgcg
      961 ccattaccga gtccgggctg cgcgttggtg cggatatctc ggtagtggga tacgacgata
     1021 ccgaagacag ctcatgttat atcccgccgt taaccaccat caaacaggat tttcgcctgc
     1081 tggggcaaac cagcgtggac cgcttgctgc aactctctca gggccaggcg gtgaagggca
     1141 atcagctgtt gcccgtctca ctggtgaaaa gaaaaaccac cctggcgccc aatacgcaaa
     1201 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
     1261 tggaaagcgg gcagtgaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt
     1321 tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgcccta
     1381 gacctagggc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacgtaa
     1441 atcactgcat aattcgtgtc gctcaaggcg cactcccgtt ctggataatg ttttttgcgc
     1501 cgacatcata acggttctgg ctaatattct gaaatgagct gttgacaatt aatcatcggc
     1561 tcgtataatg tgtggaattg tgagcggata acaatttcac acacgggttt ccggtaacaa
     1621 cacccttaag acgtaatgat ctctattcgc actaagctca gtttatggtt aacgggctta
     1681 ttggtgttag gcacagtgat tggactcata ttatttgagt ccatgctgcg ccaagcgttt
     1741 catgactcga ttatcaatcg cttagaagaa gatttagagc acattatgct ggcgacccat
     1801 ttcaataatg gggagatcca tatcgatcaa agccaactat caagctttta ccgccccgcc
     1861 tattcggggc gttatttcca gctcaacctc cccaatgaag tgatccgctc ccgttcactt
     1921 tgggatatgc agctcgatat cgagccttta gcaccaagtc aaacccgagt ttggcaggca
     1981 aagggaccca aaaacaatga catgcagtta ttatccctag gattaaattc cagcagcgcg
     2041 aatgtaaccg cgacactcac cgtggcgcag gacttgagta tcggtcgacg ggtatttagt
     2101 gaagtctacg gcactaagct tggggttaac cttgcaatgt tagtggcgat gatcgcgggg
     2161 atcttcttaa tcctgcgcca atcgtttaag cccgtgaaac aaatccaaca tgcgctatca
     2221 cgcctacaag aaggtgagat taaggccttg gagttaaaca atattccacc ggaagttcag
     2281 ccgctggcca agacctataa cgaattactg gaatacaccg ccaagcaaat tgaacgcagc
     2341 cgaaataact taggtaattt aagccacggc ctaaaaaccc cactggccgt aatgcagcaa
     2401 caggtcgaag ccttaggcct taaagatccc gaaaccgcta tggcgctgca acaacagctc
     2461 gatgcgatcc ataaaatggt cgaacgtaag ctcgccgcag cgcggatcac tggcgatatg
     2521 ttgcccgcag cgcagttagt ggtcccaagg gatctccata gcctcgccaa taccctaaac
     2581 aaagtgcata gaagcaaaaa catcagctgc aactttgagc ttgacccgag tatccagcgt
     2641 ctacccatcc atcgggaaga tggtatggag cttttaggta atctgttaga taacgccttt
     2701 aaatgggcgg atagccaagt cacagtcaaa ctttggaaag cacaaaacaa actgtatctc
     2761 agcattgatg atgatggccc aggcgttgcc gatgatgaac tggataaact cacccaaagg
     2821 ggaactcgac tcgatgaatc ggtcatgggc catggtttag gtttatcgat agtcaaggaa
     2881 atcgccgagc aatatggcat tgagcttaaa tttacccaca gccgccgatt atctggactc
     2941 tgtattgagc tggtattgag ttaataatct agaccaggca tcaaataaaa cgaaaggctc
     3001 agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctctactaga
     3061 gtcacactgg ctcaccttcg ggtgggcctt tctgcgttta taggtacccg gggatcctct
     3121 agagtcgacc tgcaggcatg caagcttagc aagcgaaccg gaattgccag ctggggcgcc
     3181 ctctggtaag gttgggaagc cctgcaaagt aaactggatg gctttcttgc cgccaaggat
     3241 ctgatggcgc aggggatcaa gatctgatca agagacagga tgaggatcgt ttcgtatgag
     3301 ggaagcggtg atcgccgaag tatcgactca actatcagag gtagttggcg tcatcgagcg
     3361 ccatctcgaa ccgacgttgc tggccgtaca tttgtacggc tccgcagtgg atggcggcct
     3421 gaagccacac agtgatattg atttgctggt tacggtgacc gtaaggcttg atgaaacaac
     3481 gcggcgagct ttgatcaacg accttttgga aacttcggct tcccctggag agagcgagat
     3541 tctccgcgct gtagaagtca ccattgttgt gcacgacgac atcattccgt ggcgttatcc
     3601 agctaagcgc gaactgcaat ttggagaatg gcagcgcaat gacattcttg caggtatctt
     3661 cgagccagcc acgatcgaca ttgatctggc tatcttgctg acaaaagcaa gagaacatag
     3721 cgttgccttg gtaggtccag cggcggagga actctttgat ccggttcctg aacaggatct
     3781 atttgaggcg ctaaatgaaa ccttaacgct atggaactcg ccgcccgact gggctggcga
     3841 tgagcgaaat gtagtgctta cgttgtcccg catttggtac agcgcagtaa ccggcaaaat
     3901 cgcgccgaag gatgtcgctg ccgactgggc aatggagcgc ctgccggccc agtatcagcc
     3961 cgtcatactt gaagctagac aggcttatct tggacaagaa gaagatcgct tggcctcgcg
     4021 cgcagatcag ttggaagaat ttgtccacta cgtgaaaggc gagatcacca aggtagtcgg
     4081 caaataagag ctcgcttgga ctcctgttga tagatccagt aatgacctca gaactccatc
     4141 tggatttgtt cagaacgctc ggttgccgcc gggcgttttt tattggtgag aatccaagca
     4201 ctagtaacaa cttatatcgt atggggctga cttcaggtgc tacatttgaa gagataaatt
     4261 gcactgaaat ctagtaatat tttatctgat taataagatg atcttcttga gatcgttttg
     4321 gtctgcgcgt aatctcttgc tctgaaaacg aaaaaaccgc cttgcagggc ggtttttcga
     4381 aggttctctg agctaccaac tctttgaacc gaggtaactg gcttggagga gcgcagtcac
     4441 caaaacttgt cctttcagtt tagccttaac cggcgcatga cttcaagact aactcctcta
     4501 aatcaattac cagtggctgc tgccagtggt gcttttgcat gtctttccgg gttggactca
     4561 agacgatagt taccggataa ggcgcagcgg tcggactgaa cggggggttc gtgcatacag
     4621 tccagcttgg agcgaactgc ctacccggaa ctgagtgtca ggcgtggaat gagacaaacg
     4681 cggccataac agcggaatga caccggtaaa ccgaaaggca ggaacaggag agcgcacgag
     4741 ggagccgcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccaccactg
     4801 atttgagcgt cagatttcgt gatgcttgtc aggggggcgg agcctatgga aaaacggctt
     4861 tgccgcggcc ctctcacttc cctgttaagt atcttcctgg catcttcc
//
