LOCUS       sequence-239123-        4727 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of torR(1-134aa)-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 4727)
  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 4727)
  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: pKD265
FEATURES             Location/Qualifiers
     source          1..4727
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      6..92
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(256..844)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(336..355)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(932..1026)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1047..1706)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1621..1640
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(1707..1809)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     1875..1893
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(1914..1932,1933..1938,1939..1955,1956..1961,
                     1962..1987)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    1914..1932
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1939..1957
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      2760..2831
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2847..2874
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             3108..3119
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             3126..3839
                     /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      3866..3937
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3953..3980
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        4032..4066
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     CDS             4101..4727
                     /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(4177..4196)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
ORIGIN
        1 ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt
       61 tgtcggtgaa cgctctcctg agtaggacaa atccgccgcc ctagacctag gcgttcggct
      121 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
      181 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
      241 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
      301 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
      361 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
      421 tctcccttcg ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc tcagttcggt
      481 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
      541 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
      601 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
      661 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
      721 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
      781 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
      841 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
      901 ttaagggatt ttggtcatga ctagtgcttg gattctcacc aataaaaaac gcccggcggc
      961 aaccgagcgt tctgaacaaa tccagatgga gttctgaggt cattactgga tctatcaaca
     1021 ggagtccaag cgagctcgat atcaaattac gccccgccct gccactcatc gcagtactgt
     1081 tgtaattcat taagcattct gccgacatgg aagccatcac agacggcatg atgaacctga
     1141 atcgccagcg gcatcagcac cttgtcgcct tgcgtataat atttgcccat ggtgaaaacg
     1201 ggggcgaaga agttgtccat attggccacg tttaaatcaa aactggtgaa actcacccag
     1261 ggattggctg agacgaaaaa catattctca ataaaccctt tagggaaata ggccaggttt
     1321 tcaccgtaac acgccacatc ttgcgaatat atgtgtagaa actgccggaa atcgtcgtgg
     1381 tattcactcc agagcgatga aaacgtttca gtttgctcat ggaaaacggt gtaacaaggg
     1441 tgaacactat cccatatcac cagctcaccg tctttcattg ccatacgaaa ttccggatga
     1501 gcattcatca ggcgggcaag aatgtgaata aaggccggat aaaacttgtg cttatttttc
     1561 tttacggtct ttaaaaaggc cgtaatatcc agctgaacgg tctggttata ggtacattga
     1621 gcaactgact gaaatgcctc aaaatgttct ttacgatgcc attgggatat atcaacggtg
     1681 gtatatccag tgattttttt ctccatttta gcttccttag ctcctgaaaa tctcgataac
     1741 tcaaaaaata cgcccggtag tgatcttatt tcattatggt gaaagttgga acctcttacg
     1801 tgccgatcaa cgtctcattt tcgccagata tcgacgtcta agaaaccatt attatcatga
     1861 cattaaccta taaaaatagg cgtatcacga ggccctttcg tcttcacctc gagtccctat
     1921 cagtgataga gattgacatc cctatcagtg atagagatac tgagcacatc agcaggacgc
     1981 actgacctaa gcgtaaaata agtaaattac acctatacaa gtatgccaca tcacattgtt
     2041 attgttgaag atgagccggt tacccaggcg cgattacaat cctacttcac tcaggagggg
     2101 tataccgttt ccgttacggc gagcggtgct gggctacggg aaattatgca gaatcagtcg
     2161 gtagatttaa ttctgctgga tatcaactta cccgatgaaa atggcctgat gttaacccgc
     2221 gccctgcgag aacgctcaac ggtggggatt attctggtta ccggacgcag cgatcggatt
     2281 gaccgtattg ttgggctgga aatgggcgca gacgattacg tcaccaaacc gctggaactg
     2341 cgcgaactgg tagtacgggt gaaaaatctg ctctggcgaa tcgacctcgc gcgacaagct
     2401 caaccgcaca ctcaggacaa ctgcgtggag cttggcgggc tgacgattta tcctgatcag
     2461 aatgaggccg aatggaacag cgtccggatt ttgttttccc aaaaagaatt ccagcttctc
     2521 tcgatttttg tgagggagca taaaaaaatt gtctctcgtg atgaactatt ggaggcgtta
     2581 tgggatgacg ttgattttgt cgatgacaat acgttaacgg tcaatgtcaa tcggctgcgg
     2641 agaaaactgg agaatgcagg actgacagat tgcatttcca caatcagagg acaaggttat
     2701 caatttcaag tcaatcggaa ggatgaagcg gaatgttaat aataatctag accaggcatc
     2761 aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg
     2821 tgaacgctct ctactagagt cacactggct caccttcggg tgggcctttc tgcgtttata
     2881 gtaaatcact gcataattcg tgtcgctcaa ggcgcactcc cgttctggat aatgtttttt
     2941 gcgccgacat cataacggtt ctggcaaata ttctgaaatg agctggttag ctagtcacac
     3001 cctcgtgaat gtgacagcat tgtaagattg gggagcggaa ttgcaagaaa gttcgatggg
     3061 aggatgctga cttccttttt ataataaaga aaaaggagga gcagaacaaa gaggagaaat
     3121 actagatgcg taaaggcgaa gagctgttca ctggtgtcgt ccctattctg gtggaactgg
     3181 atggtgatgt caacggtcat aagttttccg tgcgtggcga gggtgaaggt gacgcaacta
     3241 atggtaaact gacgctgaag ttcatctgta ctactggtaa actgccggta ccttggccga
     3301 ctctggtaac gacgctgact tatggtgttc agtgctttgc tcgttatccg gaccatatga
     3361 agcagcatga cttcttcaag tccgccatgc cggaaggcta tgtgcaggaa cgcacgattt
     3421 cctttaagga tgacggcacg tacaaaacgc gtgcggaagt gaaatttgaa ggcgataccc
     3481 tggtaaaccg cattgagctg aaaggcattg actttaaaga agacggcaat atcctgggcc
     3541 ataagctgga atacaatttt aacagccaca atgtttacat caccgccgat aaacaaaaaa
     3601 atggcattaa agcgaatttt aaaattcgcc acaacgtgga ggatggcagc gtgcagctgg
     3661 ctgatcacta ccagcaaaac actccaatcg gtgatggtcc tgttctgctg ccagacaatc
     3721 actatctgag cacgcaaagc gttctgtcta aagatccgaa cgagaaacgc gatcatatgg
     3781 ttctgctgga gttcgtaacc gcagcgggca tcacgcatgg tatggatgaa ctgtacaaat
     3841 gatgataata atctagacca ggcatcaaat aaaacgaaag gctcagtcga aagactgggc
     3901 ctttcgtttt atctgttgtt tgtcggtgaa cgctctctac tagagtcaca ctggctcacc
     3961 ttcgggtggg cctttctgcg tttataggta cccgtggatc ctctagagtc gacctgcagg
     4021 catgcaagct ttttacggct agctcagtcc taggtatagt gctagcccag ccagagaaac
     4081 actctttaac agggggtagt atgatgtctc gtttagataa aagtaaagtg attaacagcg
     4141 cattagagct gcttaatgag gtcggaatcg aaggtttaac aacccgtaaa ctcgcccaga
     4201 agctaggtgt agagcagcct acattgtatt ggcatgtaaa aaataagcgg gctttgctcg
     4261 acgccttagc cattgagatg ttagataggc accatactca cttttgccct ttagaagggg
     4321 aaagctggca agatttttta cgtaataacg ctaaaagttt tagatgtgct ttactaagtc
     4381 atcgcgatgg agcaaaagta catttaggta cacggcctac agaaaaacag tatgaaactc
     4441 tcgaaaatca attagccttt ttatgccaac aaggtttttc actagagaat gcattatatg
     4501 cactcagcgc agtggggcat tttactttag gttgcgtatt ggaagatcaa gagcatcaag
     4561 tcgctaaaga agaaagggaa acacctacta ctgatagtat gccgccatta ttacgacaag
     4621 ctatcgaatt atttgatcac caaggtgcag agccagcctt cttattcggc cttgaattga
     4681 tcatatgcgg attagaaaaa caacttaaat gtgaaagtgg gtcttaa
//
