LOCUS       sequence-247892-        4781 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of torR(1-134aa)-yxdJ(132-229aa), ompR137 chimera under 
            PLtetO-1, output Promoter PyxdJ181.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4781)
  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 4781)
  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: pKD266
FEATURES             Location/Qualifiers
     source          1..4781
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     70..88
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(109..127,128..133,134..150,151..156,157..182)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    109..127
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    134..152
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      937..1008
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1024..1051
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             1356..1367
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1374..2087
                     /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      2114..2185
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2201..2228
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        2280..2314
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     CDS             2349..2975
                     /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(2425..2444)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      2981..3067
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(3232..3820)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3312..3331)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3908..4002)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(4023..4682)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4597..4616
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 gccaacgtct cattttcgcc agatatcgac gtctaagaaa ccattattat catgacatta
       61 acctataaaa ataggcgtat cacgaggccc tttcgtcttc acctcgagtc cctatcagtg
      121 atagagattg acatccctat cagtgataga gatactgagc acatcagcag gacgcactga
      181 cctaagcgta aaataagtaa attacaccta tacaagtatg ccacatcaca ttgttattgt
      241 tgaagatgag ccggttaccc aggcgcgatt acaatcctac ttcactcagg aggggtatac
      301 cgtttccgtt acggcgagcg gtgctgggct acgggaaatt atgcagaatc agtcggtaga
      361 tttaattctg ctggatatca acttacccga tgaaaatggc ctgatgttaa cccgcgccct
      421 gcgagaacgc tcaacggtgg ggattattct ggttaccgga cgcagcgatc ggattgaccg
      481 tattgttggg ctggaaatgg gcgcagacga ttacgtcacc aaaccgctgg aactgcgcga
      541 actggtagta cgggtgaaaa atctgctctg gcgaatcgac ctcgcgcgac aagctcaacc
      601 gcacactcag gacaactgcg ttgaatatgc cggcgttcag ctctttgtgg aacggtttga
      661 actgcgtttt caggatgaaa aaagtgagct ttctaaaaaa gaaagcaagc ttttggaagt
      721 gctgcttgag cggggagaaa aggtgacgag tcgggaccgt ctcatggaaa agacgtggga
      781 caccgacata ttcatcgatg ataatacact taacgtgtat atcacgcggc tcagaaaaaa
      841 actgcgggag ctgaatgcgc ctgtttctat tgaagcggtg cggggcgaag gctaccagct
      901 gagggcgcag tcatgataat aatctagacc aggcatcaaa taaaacgaaa ggctcagtcg
      961 aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcta ctagagtcac
     1021 actggctcac cttcgggtgg gcctttctgc gtttatagta aatcactgca taattcgtgt
     1081 cgctcaaggc gcactcccgt tctggataat gttttttgcg ccgacatcat aacggttctg
     1141 gcaaatattc tgaaatgagc tggttagcta gtcaggacat cagttcgatt ttcatttctt
     1201 acaaaaatgt aaggctgcgt taagaaaaca acatagcccg ctttgtttcc ccttgataac
     1261 atggatttat gtcaagggga tttttatgtt gaacgtcccg gcactacatg aaagcagccc
     1321 agatcaacaa gaataaaacc ataaggagga ttttcaaaga ggagaaatac tagatgcgta
     1381 aaggcgaaga gctgttcact ggtgtcgtcc ctattctggt ggaactggat ggtgatgtca
     1441 acggtcataa gttttccgtg cgtggcgagg gtgaaggtga cgcaactaat ggtaaactga
     1501 cgctgaagtt catctgtact actggtaaac tgccggtacc ttggccgact ctggtaacga
     1561 cgctgactta tggtgttcag tgctttgctc gttatccgga ccatatgaag cagcatgact
     1621 tcttcaagtc cgccatgccg gaaggctatg tgcaggaacg cacgatttcc tttaaggatg
     1681 acggcacgta caaaacgcgt gcggaagtga aatttgaagg cgataccctg gtaaaccgca
     1741 ttgagctgaa aggcattgac tttaaagaag acggcaatat cctgggccat aagctggaat
     1801 acaattttaa cagccacaat gtttacatca ccgccgataa acaaaaaaat ggcattaaag
     1861 cgaattttaa aattcgccac aacgtggagg atggcagcgt gcagctggct gatcactacc
     1921 agcaaaacac tccaatcggt gatggtcctg ttctgctgcc agacaatcac tatctgagca
     1981 cgcaaagcgt tctgtctaaa gatccgaacg agaaacgcga tcatatggtt ctgctggagt
     2041 tcgtaaccgc agcgggcatc acgcatggta tggatgaact gtacaaatga tgataataat
     2101 ctagaccagg catcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat
     2161 ctgttgtttg tcggtgaacg ctctctacta gagtcacact ggctcacctt cgggtgggcc
     2221 tttctgcgtt tataggtacc cgtggatcct ctagagtcga cctgcaggca tgcaagcttt
     2281 ttacggctag ctcagtccta ggtatagtgc tagcccagcc agagaaacac tctttaacag
     2341 ggggtagtat gatgtctcgt ttagataaaa gtaaagtgat taacagcgca ttagagctgc
     2401 ttaatgaggt cggaatcgaa ggtttaacaa cccgtaaact cgcccagaag ctaggtgtag
     2461 agcagcctac attgtattgg catgtaaaaa ataagcgggc tttgctcgac gccttagcca
     2521 ttgagatgtt agataggcac catactcact tttgcccttt agaaggggaa agctggcaag
     2581 attttttacg taataacgct aaaagtttta gatgtgcttt actaagtcat cgcgatggag
     2641 caaaagtaca tttaggtaca cggcctacag aaaaacagta tgaaactctc gaaaatcaat
     2701 tagccttttt atgccaacaa ggtttttcac tagagaatgc attatatgca ctcagcgcag
     2761 tggggcattt tactttaggt tgcgtattgg aagatcaaga gcatcaagtc gctaaagaag
     2821 aaagggaaac acctactact gatagtatgc cgccattatt acgacaagct atcgaattat
     2881 ttgatcacca aggtgcagag ccagccttct tattcggcct tgaattgatc atatgcggat
     2941 tagaaaaaca acttaaatgt gaaagtgggt cttaaggcat caaataaaac gaaaggctca
     3001 gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag
     3061 gacaaatccg ccgccctaga cctagggcgt tcggctgcgg cgagcggtat cagctcactc
     3121 aaaggcggta atacggttat ccacagaatc aggggataac gcatgcaaga acatgtgagc
     3181 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     3241 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     3301 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     3361 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     3421 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     3481 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     3541 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     3601 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     3661 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     3721 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     3781 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     3841 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgactag
     3901 tgcttggatt ctcaccaata aaaaacgccc ggcggcaacc gagcgttctg aacaaatcca
     3961 gatggagttc tgaggtcatt actggatcta tcaacaggag tccaagcgag ctcgatatca
     4021 aattacgccc cgccctgcca ctcatcgcag tactgttgta attcattaag cattctgccg
     4081 acatggaagc catcacaaac ggcatgatga acctgaatcg ccagcggcat cagcaccttg
     4141 tcgccttgcg tataatattt gcccatggtg aaaacggggg cgaagaagtt gtccatattg
     4201 gccacgttta aatcaaaact ggtgaaactc acccagggat tggctgagac gaaaaacata
     4261 ttctcaataa accctttagg gaaataggcc aggttttcac cataacacgc cacatcttgc
     4321 gaatatatgt gtagaaactg ccggaaatcg tcgtggtatt cactccagag cgatgaaaac
     4381 gtttcagttt gctcatggaa aacggtgtaa caagggtgaa cactatccca tatcaccagc
     4441 tcaccgtctt tcattgccat acgaaattcc ggatgagcat tcatcaggcg ggcaagaatg
     4501 tgaataaagg ccggataaaa cttgtgctta tttttcttta cggtctttaa aaaggccgta
     4561 atatccagct gaacggtctg gttataggta cattgagcaa ctgactgaaa tgcctcaaaa
     4621 tgttctttac gatgccattg ggatatatca acggtggtat atccagtgat ttttttctcc
     4681 attttagctt ccttagctcc tgaaaatctc gataactcaa aaaatacgac cggtagtgat
     4741 cttatttcat tatggtgaaa gttggaacct cttacgggcc g
//
