LOCUS       sequence-239092-        4907 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 4907)
  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 4907)
  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..4907
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     162..179
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        218..295
                     /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             296..1378
                     /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
                     YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(315..334)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     terminator      1384..1470
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     promoter        join(1642..1647,1648..1663,1664..1670)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1678..1694
                     /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      3082..3153
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3169..3196
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             3396..4187
                     /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      4199..4293
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      join(4407..4907,1..45)
                     /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     4853..4872
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgtcagattt cgtgatgctt gtcagggggg cggagcctat ggaaaaacgg ctttgccgcg
       61 gccctctcac ttccctgtta agtatcttcc tggcatcttc caggaaatct ccgccccgtt
      121 cgtaagccat ttccgctcgc cgcagtcgaa cgaccgagcg tagcgagtca gtgagcgagg
      181 aagcggaata tatcccgaag cggcatgcat ttacgttgac accatcgaat ggtgcaaaac
      241 ctttcgcggt atggcatgat agcgcccgga agagagtcaa ttcagggtgg tgaatgtgaa
      301 accagtaacg ttatacgatg tcgcagagta tgccggtgtc tcttatcaga ccgtttcccg
      361 cgtggtgaac caggccagcc acgtttctgc gaaaacgcgg gaaaaagtgg aagcggcgat
      421 ggcggagctg aattacattc ccaaccgcgt ggcacaacaa ctggcgggca aacagtcgtt
      481 gctgattggc gttgccacct ccagtctggc cctgcacgcg ccgtcgcaaa ttgtcgcggc
      541 gattaaatct cgcgccgatc aactgggtgc cagcgtggtg gtgtcgatgg tagaacgaag
      601 cggcgtcgaa gcctgtaaag cggcggtgca caatcttctc gcgcaacgcg tcagtgggct
      661 gatcattaac tatccgctgg atgaccagga tgccattgct gtggaagctg cctgcactaa
      721 tgttccggcg ttatttcttg atgtctctga ccagacaccc atcaacagta ttattttctc
      781 ccatgaagac ggtacgcgac tgggcgtgga gcatctggtc gcattgggtc accagcaaat
      841 cgcgctgtta gcgggcccat taagttctgt ctcggcgcgt ctgcgtctgg ctggctggca
      901 taaatatctc actcgcaatc aaattcagcc gatagcggaa cgggaaggcg actggagtgc
      961 catgtccggt tttcaacaaa ccatgcaaat gctgaatgag ggcatcgttc ccactgcgat
     1021 gctggttgcc aacgatcaga tggcgctggg cgcaatgcgc gccattaccg agtccgggct
     1081 gcgcgttggt gcggatatct cggtagtggg atacgacgat accgaagaca gctcatgtta
     1141 tatcccgccg tcaaccacca tcaaacagga ttttcgcctg ctggggcaaa ccagcgtgga
     1201 ccgcttgctg caactctctc agggccaggc ggtgaagggc aatcagctgt tgcccgtctc
     1261 actggtgaaa agaaaaacca ccctggcgcc caatacgcaa accgcctctc cccgcgcgtt
     1321 ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagg
     1381 catcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg
     1441 tcggtgaacg ctctcctgag taggacaaat ccgccgccct agacctaggc gttcggctgc
     1501 ggcgagcggt atcagctcac tcaaaggcgg taatacgtaa atcactgcat aattcgtgtc
     1561 gctcaaggcg cactcccgtt ctggataatg ttttttgcgc cgacatcata acggttctgg
     1621 caaatattct gaaatgagct gttgacaatt aatcatcggc tcgtataatg tgtggaattg
     1681 tgagcggata acaatttcac acacgggttt ccggtaacaa cacccttaag acgtaatgat
     1741 ctctattcgc actaagctca gtttatggtt aacgggctta ttggtgttag gcacagtgat
     1801 tggactcata ttatttgagt ccatgctgcg ccaagcgttt catgactcga ttatcaatcg
     1861 cttagaagaa gatttagagc acattatgct ggcgacccat ttcaataatg gggagatcca
     1921 tatcgatcaa agccaactat caagctttta ccgccccgcc tattcggggc gttatttcca
     1981 gctcaacctc cccaatgaag tgatccgctc ccgttcactt tgggatatgc agctcgatat
     2041 cgagccttta gcaccaagtc aaacccgagt ttggcaggca aagggaccca aaaacaatga
     2101 catgcagtta ttatccctag gattaaattc cagcagcgcg aatgtaaccg cgacactcac
     2161 cgtggcgcag gacttgagta tcggtcgacg ggtatttagt gaagtctacg gcactaagct
     2221 tggggttaac cttgcaatgt tagtggcgat gatcgcgggg atcttcttaa tcctgcgcca
     2281 atcgtttaag cccgtgaaac aaatccaaca tgcgctatca cgcctacaag aaggtgagat
     2341 taaggccttg gagttaaaca atattccacc ggaagttcag ccgctggcca agacctataa
     2401 cgaattactg gaatacaccg ccaagcaaat tgaacgcagc cgaaataact taggtaattt
     2461 aagccacggc ctaaaaaccc cactggccgt aatgcagcaa caggtcgaag ccttaggcct
     2521 taaagatccc gaaaccgcta tggcgctgca acaacagctc gatgcgatcc ataaaatggt
     2581 cgaacgtaag ctcgccgcag cgcggatcac tggcgatatg ttgcccgcag cgcagttagt
     2641 ggtcccaagg gatctccata gcctcgccaa taccctaaac aaagtgcata gaagcaaaaa
     2701 catcagctgc aactttgagc ttgacccgag tatccagcgt ctacccatcc atcgggaaga
     2761 tggtatggag cttttaggta atctgttaga taacgccttt aaatgggcgg atagccaagt
     2821 cacagtcaaa ctttggaaag cacaaaacaa actgtatctc agcattgatg atgatggccc
     2881 aggcgttgcc gatgatgaac tggataaact cacccaaagg ggaactcgac tcgatgaatc
     2941 ggtcatgggc catggtttag gtttatcgat agtcaaggaa atcgccgagc aatatggcat
     3001 tgagcttaaa tttacccaca gccgccgatt atctggactc tgtattgagc tggtattgag
     3061 ttaataatct agaccaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt
     3121 cgttttatct gttgtttgtc ggtgaacgct ctctactaga gtcacactgg ctcaccttcg
     3181 ggtgggcctt tctgcgttta taggtacccg gggatcctct agagtcgacc tgcaggcatg
     3241 caagcttagc aagcgaaccg gaattgccag ctggggcgcc ctctggtaag gttgggaagc
     3301 cctgcaaagt aaactggatg gctttcttgc cgccaaggat ctgatggcgc aggggatcaa
     3361 gatctgatca agagacagga tgaggatcgt ttcgcatgag ggaagcggtg atcgccgaag
     3421 tatcgactca actatcagag gtagttggcg tcatcgagcg ccatctcgaa ccgacgttgc
     3481 tggccgtaca tttgtacggc tccgcagtgg atggcggcct gaagccacac agtgatattg
     3541 atttgctggt tacggtgacc gtaaggcttg atgaaacaac gcggcgagct ttgatcaacg
     3601 accttttgga aacttcggct tcccctggag agagcgagat tctccgcgct gtagaagtca
     3661 ccattgttgt gcacgacgac atcattccgt ggcgttatcc agctaagcgc gaactgcaat
     3721 ttggagaatg gcagcgcaat gacattcttg caggtatctt cgagccagcc acgatcgaca
     3781 ttgatctggc tatcttgctg acaaaagcaa gagaacatag cgttgccttg gtaggtccag
     3841 cggcggagga actctttgat ccggttcctg aacaggatct atttgaggcg ctaaatgaaa
     3901 ccttaacgct atggaactcg ccgcccgact gggctggcga tgagcgaaat gtagtgctta
     3961 cgttgtcccg catttggtac agcgcagtaa ccggcaaaat cgcgccgaag gatgtcgctg
     4021 ccgactgggc aatggagcgc ctgccggccc agtatcagcc cgtcatactt gaagctagac
     4081 aggcttatct tggacaagaa gaagatcgct tggcctcgcg cgcagatcag ttggaagaat
     4141 ttgtccacta cgtgaaaggc gagatcacca aggtagtcgg caaataagag ctcgcttgga
     4201 ctcctgttga tagatccagt aatgacctca gaactccatc tggatttgtt cagaacgctc
     4261 ggttgccgcc gggcgttttt tattggtgag aatccaagca ctagtaacaa cttatatcgt
     4321 atggggctga cttcaggtgc tacatttgaa gagataaatt gcactgaaat ctagtaatat
     4381 tttatctgat taataagatg atcttcttga gatcgttttg gtctgcgcgt aatctcttgc
     4441 tctgaaaacg aaaaaaccgc cttgcagggc ggtttttcga aggttctctg agctaccaac
     4501 tctttgaacc gaggtaactg gcttggagga gcgcagtcac caaaacttgt cctttcagtt
     4561 tagccttaac cggcgcatga cttcaagact aactcctcta aatcaattac cagtggctgc
     4621 tgccagtggt gcttttgcat gtctttccgg gttggactca agacgatagt taccggataa
     4681 ggcgcagcgg tcggactgaa cggggggttc gtgcatacag tccagcttgg agcgaactgc
     4741 ctacccggaa ctgagtgtca ggcgtggaat gagacaaacg cggccataac agcggaatga
     4801 caccggtaaa ccgaaaggca ggaacaggag agcgcacgag ggagccgcca gggggaaacg
     4861 cctggtatct ttatagtcct gtcgggtttc gccaccactg atttgag
//
