LOCUS       sequence-245431-        5084 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of SO_4387 under Ptac.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5084)
  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 5084)
  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: pKD280
FEATURES             Location/Qualifiers
     source          1..5084
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      81..167
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     promoter        join(340..345,346..361,362..368)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    376..392
                     /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      1956..2027
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2043..2070
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             2270..3061
                     /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      3073..3167
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      3281..3826
                     /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     3727..3746
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3943..3960
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        3999..4076
                     /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(4077..5084,1..75)
                     /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(4096..4115)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
       61 gaaagcgggc agtgaggcat caaataaaac gaaaggctca gtcgaaagac tgggcctttc
      121 gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg ccgccctaga
      181 cctagggcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacgtaaat
      241 cactgcataa ttcgtgtcgc tcaaggcgca ctcccgttct ggataatgtt ttttgcgccg
      301 acatcataac ggttctggct aatattctga aatgagctgt tgacaattaa tcatcggctc
      361 gtataatgtg tggaattgtg agcggataac aatttcacac aaaaaattaa gttctatatt
      421 acagcttata gtagaatgac acagaattat gaacactacc tttttctatt tgttatcatc
      481 actggcctga taatgggagt tattagttat tttggctttc gagcgctgac acaagaaagc
      541 accctgaatg aataccaaac tcaacaattg gctaaaagcc acgtagctca aactcaatca
      601 tttattctgc aacagctgaa tcaaaaacag gttagattag atgccatgct agcctatctg
      661 tctctagata aaaatgcgct acataccttg atatcccaag acagcgatat cgagaacatc
      721 tttgttctgc aaaaaaataa gctcgctttt cctgatatat ctcaaccttt aagtttgaaa
      781 gatcaaacgc tggctgaact gattatgcca ataagccatg accctacatt gttaatggct
      841 aaacaaccca catctgaaga taaaagccct aactctggct ggttagtctt acaagacaat
      901 aactatccag cacttattta ctggcgtaaa accgatgaag atacggtagg tataaaactc
      961 tcttacgtta aatttttagc tgatgttatc tctgctctcg atacaaattt tacacctgat
     1021 ggtataaaaa tttccgataa cggccaggta ctttaccaac tgtccccaga gcctatcggt
     1081 aataatcaaa cgccttctta tagccagaat ttaccttttc cgctacacgg ttggcagatt
     1141 gactattata ccgcacctgt gaaagggcaa tctctttact atctaggcac catattaatt
     1201 gtcaccatta ttttggccgt tggctttatg gcatttatgc tatatcgcga gtttaatcgc
     1261 gcaaccagat tagcaagcca acaggttagt tttgtcggac aggtttctca cgaacttaaa
     1321 acgccactga cgaatatcac gctttatgct gaaatgctta acgaaatgga agcagaggaa
     1381 aacgaccaga gagcccatta tcttggagtc attatcagtg aaagccaacg cctatctcgc
     1441 ctgattcaaa acgtgttgac cttcaccaaa ctgccacaaa tttgcattcg ccaagtcgat
     1501 attaatcaat taatgaatca aatttatact gcgttcaaac ccgtatttga agccaaagaa
     1561 atccatctca acctcaatat taatgggcaa ttatcaacga aaacagacgt tgatctggta
     1621 acccagatta tcagtaacct actcagtaat gccgaaaaat atgctgcaac aggtaagcag
     1681 gttgacctta atgtcgaaca agataacgac aatattttta tacatatccg agactacggt
     1741 aacggcataa cggctaaaga gttaaaacaa atattccagc cgttttatcg ggttagatca
     1801 tcgattaccg aaggagtgag cggaacaggt atcggactca ccattgcccg ccagttagct
     1861 gaaacgctga gcggtgagat taacgcaatc aacaatgagc ccggaatgac gtttacgcta
     1921 agacttccaa aacgctaata atctagacca ggcatcaaat aaaacgaaag gctcagtcga
     1981 aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctctac tagagtcaca
     2041 ctggctcacc ttcgggtggg cctttctgcg tttataggta cccggggatc ctctagagtc
     2101 gacctgcagg catgcaagct tagcaagcga accggaattg ccagctgggg cgccctctgg
     2161 taaggttggg aagccctgca aagtaaactg gatggctttc ttgccgccaa ggatctgatg
     2221 gcgcagggga tcaagatctg atcaagagac aggatgagga tcgtttcgta tgagggaagc
     2281 ggtgatcgcc gaagtatcga ctcaactatc agaggtagtt ggcgtcatcg agcgccatct
     2341 cgaaccgacg ttgctggccg tacatttgta cggctccgca gtggatggcg gcctgaagcc
     2401 acacagtgat attgatttgc tggttacggt gaccgtaagg cttgatgaaa caacgcggcg
     2461 agctttgatc aacgaccttt tggaaacttc ggcttcccct ggagagagcg agattctccg
     2521 cgctgtagaa gtcaccattg ttgtgcacga cgacatcatt ccgtggcgtt atccagctaa
     2581 gcgcgaactg caatttggag aatggcagcg caatgacatt cttgcaggta tcttcgagcc
     2641 agccacgatc gacattgatc tggctatctt gctgacaaaa gcaagagaac atagcgttgc
     2701 cttggtaggt ccagcggcgg aggaactctt tgatccggtt cctgaacagg atctatttga
     2761 ggcgctaaat gaaaccttaa cgctatggaa ctcgccgccc gactgggctg gcgatgagcg
     2821 aaatgtagtg cttacgttgt cccgcatttg gtacagcgca gtaaccggca aaatcgcgcc
     2881 gaaggatgtc gctgccgact gggcaatgga gcgcctgccg gcccagtatc agcccgtcat
     2941 acttgaagct agacaggctt atcttggaca agaagaagat cgcttggcct cgcgcgcaga
     3001 tcagttggaa gaatttgtcc actacgtgaa aggcgagatc accaaggtag tcggcaaata
     3061 agagctcgct tggactcctg ttgatagatc cagtaatgac ctcagaactc catctggatt
     3121 tgttcagaac gctcggttgc cgccgggcgt tttttattgg tgagaatcca agcactagta
     3181 acaacttata tcgtatgggg ctgacttcag gtgctacatt tgaagagata aattgcactg
     3241 aaatctagta atattttatc tgattaataa gatgatcttc ttgagatcgt tttggtctgc
     3301 gcgtaatctc ttgctctgaa aacgaaaaaa ccgccttgca gggcggtttt tcgaaggttc
     3361 tctgagctac caactctttg aaccgaggta actggcttgg aggagcgcag tcaccaaaac
     3421 ttgtcctttc agtttagcct taaccggcgc atgacttcaa gactaactcc tctaaatcaa
     3481 ttaccagtgg ctgctgccag tggtgctttt gcatgtcttt ccgggttgga ctcaagacga
     3541 tagttaccgg ataaggcgca gcggtcggac tgaacggggg gttcgtgcat acagtccagc
     3601 ttggagcgaa ctgcctaccc ggaactgagt gtcaggcgtg gaatgagaca aacgcggcca
     3661 taacagcgga atgacaccgg taaaccgaaa ggcaggaaca ggagagcgca cgagggagcc
     3721 gccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc actgatttga
     3781 gcgtcagatt tcgtgatgct tgtcaggggg gcggagccta tggaaaaacg gctttgccgc
     3841 ggccctctca cttccctgtt aagtatcttc ctggcatctt ccgggaaatc tccgccccgt
     3901 tcgtaagcca tttccgctcg ccgcagtcga acgaccgagc gtagcgagtc agtgagcgag
     3961 gaagcggaat atatcccgaa gcggcatgca tttacgttga caccatcgaa tggtgcaaaa
     4021 cctttcgcgg tatggcatga tagcgcccgg aagagagtca attcagggtg gtgaatgtga
     4081 aaccagtaac gttatacgat gtcgcagagt atgccggtgt ctcttatcag accgtttccc
     4141 gcgtggtgaa ccaggccagc cacgtttctg cgaaaacgcg ggaaaaagtg gaagcggcga
     4201 tggcggagct gaattacatt cccaaccgcg tggcacaaca actggcgggc aaacagtcgt
     4261 tgctgattgg cgttgccacc tccagtctgg ccctgcacgc gccgtcgcaa attgtcgcgg
     4321 cgattaaatc tcgcgccgat caactgggtg ccagcgtggt ggtgtcgatg gtagaacgaa
     4381 gcggcgtcga agcctgtaaa gcggcggtgc acaatcttct cgcgcaacgc gtcagtgggc
     4441 tgatcattaa ctatccgctg gatgaccagg atgccattgc tgtggaagct gcctgcacta
     4501 atgttccggc gttatttctt gatgtctctg accagacacc catcaacagt attattttct
     4561 cccatgaaga cggtacgcga ctgggcgtgg agcatctggt cgcattgggt caccagcaaa
     4621 tcgcgctgtt agcgggccca ttaagttctg tctcggcgcg tctgcgtctg gctggctggc
     4681 ataaatatct cactcgcaat caaattcagc cgatagcgga acgggaaggc gactggagtg
     4741 ccatgtccgg ttttcaacaa accatgcaaa tgctgaatga gggcatcgtt cccactgcga
     4801 tgctggttgc caacgatcag atggcgctgg gcgcaatgcg cgccattacc gagtccgggc
     4861 tgcgcgttgg tgcggatatc tcggtagtgg gatacgacga taccgaagac agctcatgtt
     4921 atatcccgcc gttaaccacc atcaaacagg attttcgcct gctggggcaa accagcgtgg
     4981 accgcttgct gcaactctct cagggccagg cggtgaaggg caatcagctg ttgcccgtct
     5041 cactggtgaa aagaaaaacc accctggcgc ccaatacgca aacc
//
