LOCUS       sequence-245241-        4716 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of SO_4388(1-130aa)-psdR(134-237aa), ompR137 chimera 
            under PLtetO-1, output Promoter PpsdA110.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4716)
  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 4716)
  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: pKD279
FEATURES             Location/Qualifiers
     source          1..4716
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     71..89
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(110..128,129..134,135..151,152..157,158..183)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    110..128
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    135..153
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      944..1015
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1031..1058
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             1292..1303
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1310..2023
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      2050..2121
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2137..2164
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        2216..2250
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     CDS             2285..2911
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MMSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWH
                     VKNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGT
                     RPTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPT
                     TDSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     complement(2361..2380)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      2917..3003
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(3168..3756)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3248..3267)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3844..3938)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3959..4618)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4533..4552
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 ggccaacgtc tcattttcgc cagatatcga cgtctaagaa accattatta tcatgacatt
       61 aacctataaa aataggcgta tcacgaggcc ctttcgtctt cacctcgagt ccctatcagt
      121 gatagagatt gacatcccta tcagtgatag agatactgag cacatcagca ggacgcactg
      181 acctgccccg tataataggt tataagacgg aattatttat gaaaattctt atcgccgaag
      241 atgatattca tatccgccaa gggttggccg atatgctttc cagagaggga tattccgttc
      301 tgcttgctga taatggcaaa gttgccctgt taaaatatca acaagagcag ccggatttta
      361 ttattctgga tattatgatg cctgagttag atggctattc agtctgcaaa gaaatcagaa
      421 agcacgatga acaaacgccg gtgatctttt tatccgctaa aggtgaagag cttgataaag
      481 ttctggggct tgagcttggt gcggatgact atattaataa accatttggt attcacgagg
      541 ttcgggcgcg aatcaaaaca attgcccgac gctgtctgaa agccaaacag aattcaccag
      601 atcaaagcgt ggagcttggc gggctgacga tttatcctga tcagaatgag gccgaatgga
      661 acagcgtccg gattttgttt tcccaaaaag aattccagct tctctcgatt tttgtgaggg
      721 agcataaaaa aattgtctct cgtgatgaac tattggaggc gttatgggat gacgttgatt
      781 ttgtcgatga caatacgtta acggtcaatg tcaatcggct gcggagaaaa ctggagaatg
      841 caggactgac agattgcatt tccacaatca gaggacaagg ttatcaattt caagtcaatc
      901 ggaaggatga agcggaatgt taataataat ctagaccagg catcaaataa aacgaaaggc
      961 tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctctacta
     1021 gagtcacact ggctcacctt cgggtgggcc tttctgcgtt tatagtaaat cactgcataa
     1081 ttcgtgtcgc tcaaggcgca ctcccgttct ggataatgtt ttttgcgccg acatcataac
     1141 ggttctggca aatattctga aatgagctgg ttagctagtc acaccctcgt gaatgtgaca
     1201 gcattgtaag attggggagc ggaattgcaa gaaagttcga tgggaggatg ctgacttcct
     1261 ttttataata aagaaaaagg aggagcagaa caaagaggag aaatactaga tgcgtaaagg
     1321 cgaagagctg ttcactggtg tcgtccctat tctggtggaa ctggatggtg atgtcaacgg
     1381 tcataagttt tccgtgcgtg gcgagggtga aggtgacgca actaatggta aactgacgct
     1441 gaagttcatc tgtactactg gtaaactgcc ggtaccttgg ccgactctgg taacgacgct
     1501 gacttatggt gttcagtgct ttgctcgtta tccggaccat atgaagcagc atgacttctt
     1561 caagtccgcc atgccggaag gctatgtgca ggaacgcacg atttccttta aggatgacgg
     1621 cacgtacaaa acgcgtgcgg aagtgaaatt tgaaggcgat accctggtaa accgcattga
     1681 gctgaaaggc attgacttta aagaagacgg caatatcctg ggccataagc tggaatacaa
     1741 ttttaacagc cacaatgttt acatcaccgc cgataaacaa aaaaatggca ttaaagcgaa
     1801 ttttaaaatt cgccacaacg tggaggatgg cagcgtgcag ctggctgatc actaccagca
     1861 aaacactcca atcggtgatg gtcctgttct gctgccagac aatcactatc tgagcacgca
     1921 aagcgttctg tctaaagatc cgaacgagaa acgcgatcat atggttctgc tggagttcgt
     1981 aaccgcagcg ggcatcacgc atggtatgga tgaactgtac aaatgatgat aataatctag
     2041 accaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt
     2101 tgtttgtcgg tgaacgctct ctactagagt cacactggct caccttcggg tgggcctttc
     2161 tgcgtttata ggtacccgtg gatcctctag agtcgacctg caggcatgca agctttttac
     2221 ggctagctca gtcctaggta tagtgctagc ccagccagag aaacactctt taacaggggg
     2281 tagtatgatg tctcgtttag ataaaagtaa agtgattaac agcgcattag agctgcttaa
     2341 tgaggtcgga atcgaaggtt taacaacccg taaactcgcc cagaagctag gtgtagagca
     2401 gcctacattg tattggcatg taaaaaataa gcgggctttg ctcgacgcct tagccattga
     2461 gatgttagat aggcaccata ctcacttttg ccctttagaa ggggaaagct ggcaagattt
     2521 tttacgtaat aacgctaaaa gttttagatg tgctttacta agtcatcgcg atggagcaaa
     2581 agtacattta ggtacacggc ctacagaaaa acagtatgaa actctcgaaa atcaattagc
     2641 ctttttatgc caacaaggtt tttcactaga gaatgcatta tatgcactca gcgcagtggg
     2701 gcattttact ttaggttgcg tattggaaga tcaagagcat caagtcgcta aagaagaaag
     2761 ggaaacacct actactgata gtatgccgcc attattacga caagctatcg aattatttga
     2821 tcaccaaggt gcagagccag ccttcttatt cggccttgaa ttgatcatat gcggattaga
     2881 aaaacaactt aaatgtgaaa gtgggtctta aggcatcaaa taaaacgaaa ggctcagtcg
     2941 aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcct gagtaggaca
     3001 aatccgccgc cctagaccta gggcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     3061 gcggtaatac ggttatccac agaatcaggg gataacgcat gcaagaacat gtgagcaaaa
     3121 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     3181 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     3241 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     3301 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     3361 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     3421 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     3481 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     3541 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     3601 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     3661 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     3721 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     3781 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gactagtgct
     3841 tggattctca ccaataaaaa acgcccggcg gcaaccgagc gttctgaaca aatccagatg
     3901 gagttctgag gtcattactg gatctatcaa caggagtcca agcgagctcg atatcaaatt
     3961 acgccccgcc ctgccactca tcgcagtact gttgtaattc attaagcatt ctgccgacat
     4021 ggaagccatc acaaacggca tgatgaacct gaatcgccag cggcatcagc accttgtcgc
     4081 cttgcgtata atatttgccc atggtgaaaa cgggggcgaa gaagttgtcc atattggcca
     4141 cgtttaaatc aaaactggtg aaactcaccc agggattggc tgagacgaaa aacatattct
     4201 caataaaccc tttagggaaa taggccaggt tttcaccata acacgccaca tcttgcgaat
     4261 atatgtgtag aaactgccgg aaatcgtcgt ggtattcact ccagagcgat gaaaacgttt
     4321 cagtttgctc atggaaaacg gtgtaacaag ggtgaacact atcccatatc accagctcac
     4381 cgtctttcat tgccatacga aattccggat gagcattcat caggcgggca agaatgtgaa
     4441 taaaggccgg ataaaacttg tgcttatttt tctttacggt ctttaaaaag gccgtaatat
     4501 ccagctgaac ggtctggtta taggtacatt gagcaactga ctgaaatgcc tcaaaatgtt
     4561 ctttacgatg ccattgggat atatcaacgg tggtatatcc agtgattttt ttctccattt
     4621 tagcttcctt agctcctgaa aatctcgata actcaaaaaa tacgaccggt agtgatctta
     4681 tttcattatg gtgaaagttg gaacctctta cgggcc
//
