LOCUS       sequence-245247-        4705 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of SO_4637(1-127aa)-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 4705)
  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 4705)
  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: pKD287
FEATURES             Location/Qualifiers
     source          1..4705
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     62..80
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(101..119,120..125,126..142,143..148,149..174)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    101..119
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    126..144
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      924..995
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1011..1038
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             1272..1283
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1290..2003
                     /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      2030..2101
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2117..2144
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        2196..2230
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     CDS             2265..2891
                     /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(2341..2360)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      2897..2983
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(3148..3736)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3228..3247)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3824..3918)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3939..4598)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4513..4532
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 ctcattttcg ccagatatcg acgtctaaga aaccattatt atcatgacat taacctataa
       61 aaataggcgt atcacgaggc cctttcgtct tcacctcgag tccctatcag tgatagagat
      121 tgacatccct atcagtgata gagatactga gcacatcagc aggacgcact gacccgagcg
      181 acccgtatac ccttttagca cgtatatatg aaagtattaa ttgttgatga caatcatgat
      241 gttatagaaa caatcatgga ctatttaacc ctcgaaggca ttattgctga ctgcgcctat
      301 cacggtgaaa gcgccatcaa tttaatacag caaaaccatt atgacgtgat cattatggat
      361 attatgatgc ccaaattaga cggtatccgt acggtcaaaa aactgcgcga agagctgttt
      421 tgtaatacgc cgattttgtt tttaaccgcc agagatagcc tgcaggacaa aattgactca
      481 tttacctcgg gcggtgatga ctttttaaat aaacccttcg ccatggaaga actgtgtctg
      541 cgtctacggt cactgcataa tcgcgggccg cgtttagatg taggcatagt ggagcttggc
      601 gggctgacga tttatcctga tcagaatgag gccgaatgga acagcgtccg gattttgttt
      661 tcccaaaaag aattccagct tctctcgatt tttgtgaggg agcataaaaa aattgtctct
      721 cgtgatgaac tattggaggc gttatgggat gacgttgatt ttgtcgatga caatacgtta
      781 acggtcaatg tcaatcggct gcggagaaaa ctggagaatg caggactgac agattgcatt
      841 tccacaatca gaggacaagg ttatcaattt caagtcaatc ggaaggatga agcggaatgt
      901 taataataat ctagaccagg catcaaataa aacgaaaggc tcagtcgaaa gactgggcct
      961 ttcgttttat ctgttgtttg tcggtgaacg ctctctacta gagtcacact ggctcacctt
     1021 cgggtgggcc tttctgcgtt tatagtaaat cactgcataa ttcgtgtcgc tcaaggcgca
     1081 ctcccgttct ggataatgtt ttttgcgccg acatcataac ggttctggca aatattctga
     1141 aatgagctgg ttagctagtc acaccctcgt gaatgtgaca gcattgtaag attggggagc
     1201 ggaattgcaa gaaagttcga tgggaggatg ctgacttcct ttttataata aagaaaaagg
     1261 aggagcagaa caaagaggag aaatactaga tgcgtaaagg cgaagagctg ttcactggtg
     1321 tcgtccctat tctggtggaa ctggatggtg atgtcaacgg tcataagttt tccgtgcgtg
     1381 gcgagggtga aggtgacgca actaatggta aactgacgct gaagttcatc tgtactactg
     1441 gtaaactgcc ggtaccttgg ccgactctgg taacgacgct gacttatggt gttcagtgct
     1501 ttgctcgtta tccggaccat atgaagcagc atgacttctt caagtccgcc atgccggaag
     1561 gctatgtgca ggaacgcacg atttccttta aggatgacgg cacgtacaaa acgcgtgcgg
     1621 aagtgaaatt tgaaggcgat accctggtaa accgcattga gctgaaaggc attgacttta
     1681 aagaagacgg caatatcctg ggccataagc tggaatacaa ttttaacagc cacaatgttt
     1741 acatcaccgc cgataaacaa aaaaatggca ttaaagcgaa ttttaaaatt cgccacaacg
     1801 tggaggatgg cagcgtgcag ctggctgatc actaccagca aaacactcca atcggtgatg
     1861 gtcctgttct gctgccagac aatcactatc tgagcacgca aagcgttctg tctaaagatc
     1921 cgaacgagaa acgcgatcat atggttctgc tggagttcgt aaccgcagcg ggcatcacgc
     1981 atggtatgga tgaactgtac aaatgatgat aataatctag accaggcatc aaataaaacg
     2041 aaaggctcag tcgaaagact gggcctttcg ttttatctgt tgtttgttgg tgaacgctct
     2101 ctactagagt cacactggct caccttcggg tgggcctttc tgcgtttata ggtacccgtg
     2161 gatcctctag agtcgacctg caggcatgca agctttttac ggctagctca gtcctaggta
     2221 tagtgctagc ccagccagag aaacactctt taacaggggg tagtatgatg tctcgtttag
     2281 ataaaagtaa agtgattaac agcgcattag agctgcttaa tgaggtcgga atcgaaggtt
     2341 taacaacccg taaactcgcc cagaagctag gtgtagagca gcctacattg tattggcatg
     2401 taaaaaataa gcgggctttg ctcgacgcct tagccattga gatgttagat aggcaccata
     2461 ctcacttttg ccctttagaa ggggaaagct ggcaagattt tttacgtaat aacgctaaaa
     2521 gttttagatg tgctttacta agtcatcgcg atggagcaaa agtacattta ggtacacggc
     2581 ctacagaaaa acagtatgaa actctcgaaa atcaattagc ctttttatgc caacaaggtt
     2641 tttcactaga gaatgcatta tatgcactca gcgcagtggg gcattttact ttaggttgcg
     2701 tattggaaga tcaagagcat caagtcgcta aagaagaaag ggaaacacct actactgata
     2761 gtatgccgcc attattacga caagctatcg aattatttga tcaccaaggt gcagagccag
     2821 ccttcttatt cggccttgaa ttgatcatat gcggattaga aaaacaactt aaatgtgaaa
     2881 gtgggtctta aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt
     2941 tatctgttgt ttgtcggtga acgctctcct gagtaggaca aatccgccgc cctagaccta
     3001 gggcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     3061 agaatcaggg gataacgcat gcaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     3121 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     3181 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     3241 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     3301 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     3361 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     3421 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     3481 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     3541 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     3601 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     3661 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     3721 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     3781 cgaaaactca cgttaaggga ttttggtcat gactagtgct tggattctca ccaataaaaa
     3841 acgcccggcg gcaaccgagc gttctgaaca aatccagatg gagttctgag gtcattactg
     3901 gatctatcaa caggagtcca agcgagctcg atatcaaatt acgccccgcc ctgccactca
     3961 tcgcagtact gttgtaattc attaagcatt ctgccgacat ggaagccatc acaaacggca
     4021 tgatgaacct gaatcgccag cggcatcagc accttgtcgc cttgcgtata atatttgccc
     4081 atggtgaaaa cgggggcgaa gaagttgtcc atattggcca cgtttaaatc aaaactggtg
     4141 aaactcaccc agggattggc tgagacgaaa aacatattct caataaaccc tttagggaaa
     4201 taggccaggt tttcaccata acacgccaca tcttgcgaat atatgtgtag aaactgccgg
     4261 aaatcgtcgt ggtattcact ccagagcgat gaaaacgttt cagtttgctc atggaaaacg
     4321 gtgtaacaag ggtgaacact atcccatatc accagctcac cgtctttcat tgccatacga
     4381 aattccggat gagcattcat caggcgggca agaatgtgaa taaaggccgg ataaaacttg
     4441 tgcttatttt tctttacggt ctttaaaaag gccgtaatat ccagctgaac ggtctggtta
     4501 taggtacatt gagcaactga ctgaaatgcc tcaaaatgtt ctttacgatg ccattgggat
     4561 atatcaacgg tggtatatcc agtgattttt ttctccattt tagcttcctt agctcctgaa
     4621 aatctcgata actcaaaaaa tacgaccggt agtgatctta tttcattatg gtgaaagttg
     4681 gaacctctta cgggccggcc aacgt
//
