LOCUS       sequence-245194-        4840 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of ompR(1-142aa)-ccaR(132-234aa) chimera under PLtetO-1, 
            output Promoter PcpcG2-172.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4840)
  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 4840)
  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: pSR40.30
FEATURES             Location/Qualifiers
     source          1..4840
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     46..64
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(85..103,104..109,110..126,127..132,133..158)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    85..103
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    110..128
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      952..1023
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1039..1066
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             1362..1373
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1380..2093
                     /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
                     TYKTRAEVKFEGDTMVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      2120..2191
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2207..2234
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        2286..2320
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     CDS             2355..2981
                     /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(2431..2450)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      2987..3073
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(3238..3826)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3318..3337)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3914..4008)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(4029..4688)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4603..4622
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 atcgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
       61 aggccctttc gtcttcacct cgagtcccta tcagtgatag agattgacat ccctatcagt
      121 gatagagata ctgagcacat cagcaggacg cactgaccgc ataaaggact tagcaaagcc
      181 caattttaaa caaatgcaag agaactacaa gattctggtg gtcgatgacg acatgcgcct
      241 gcgtgcgctg ctggaacgtt atctcaccga acaaggcttc caggttcgaa gcgtcgctaa
      301 tgcagaacag atggatcgcc tgctgactcg tgaatctttc catcttatgg tactggattt
      361 aatgttacct ggtgaagatg gcttgtcgat ttgccgacgt cttcgtagtc agagcaaccc
      421 gatgccgatc attatggtga cggcgaaagg ggaagaagtg gaccgtatcg taggcctgga
      481 gattggcgct gacgactaca ttccaaaacc gtttaacccg cgtgaactgc tggcccgtat
      541 ccgtgcggtg ctgcgtcgtc aggcgaacga actgccaggc gcaccgtcac aggaagaggc
      601 ggtaattgct ttcggtaaga tcaggttgga tccaagcacc tatgaagtta gttatgacaa
      661 tgaggttttg tctttgaccc gcaaggaata cagcattctg gaattactac tccgcaatgg
      721 ccgtcgggtg ctaagtcgga gcatgattat cgatagtatc tggaagttgg agagtccccc
      781 agaggaagat acggttaagg tgcatgtgcg gagtttgcga caaaaattaa aaagtgccgg
      841 tttatcagca gatgccattg aaacggtcca tggcattggg tatcgtctgg ccaatttaac
      901 ggaaaaatct ttgtgccaag ggaaaaacta gtaataatct agaccaggca tcaaataaaa
      961 cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct
     1021 ctctactaga gtcacactgg ctcaccttcg ggtgggcctt tctgcgttta tagtaaatca
     1081 ctgcataatt cgtgtcgctc aaggcgcact cccgttctgg ataatgtttt ttgcgccgac
     1141 atcataacgg ttctggcaaa tattctgaaa tgagctggtt agctagtcaa gcccattgtg
     1201 cttttctcta tcaacctcag cttacctgaa ggggtgaaca ggtctgggtt aattcatgtt
     1261 gcgaaatgta acagttttag tcgcatcagc taactttccg atttctttac gattttctcc
     1321 cccttttctt caattttact ttgttaggat cgcattttta aaaagaggag aaatactaga
     1381 tgcgtaaagg cgaagagctg ttcactggtg tcgtccctat tctggtggaa ctggatggtg
     1441 atgtcaacgg tcataagttt tccgtgcgtg gcgagggtga aggtgacgca actaatggta
     1501 aactgacgct gaagttcatc tgtactactg gtaaactgcc ggtaccttgg ccgactctgg
     1561 taacgacgct gacttatggt gttcagtgct ttgctcgtta tccggaccat atgaagcagc
     1621 atgacttctt caagtccgcc atgccggaag gctatgtgca ggaacgcacg atttccttta
     1681 aggatgacgg cacgtacaaa acgcgtgcgg aagtgaaatt tgaaggcgat accatggtaa
     1741 accgcattga gctgaaaggc attgacttta aagaagacgg caatatcctg ggccataagc
     1801 tggaatacaa ttttaacagc cacaatgttt acatcaccgc cgataaacaa aaaaatggca
     1861 ttaaagcgaa ttttaaaatt cgccacaacg tggaggatgg cagcgtgcag ctggctgatc
     1921 actaccagca aaacactcca atcggtgatg gtcctgttct gctgccagac aatcactatc
     1981 tgagcacgca aagcgttctg tctaaagatc cgaacgagaa acgcgatcat atggttctgc
     2041 tggagttcgt aaccgcagcg ggcatcacgc atggtatgga tgaactgtac aaatgatgat
     2101 aataatctag accaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg
     2161 ttttatctgt tgtttgtcgg tgaacgctct ctactagagt cacactggct caccttcggg
     2221 tgggcctttc tgcgtttata ggtacccgtg gatcctctag agtcgacctg caggcatgca
     2281 agctttttac ggctagctca gtcctaggta tagtgctagc ccagccagag aaacactctt
     2341 taacaggggg tagtatgatg tctcgtttag ataaaagtaa agtgattaac agcgcattag
     2401 agctgcttaa tgaggtcgga atcgaaggtt taacaacccg taaactcgcc cagaagctag
     2461 gtgtagagca gcctacattg tattggcatg taaaaaataa gcgggctttg ctcgacgcct
     2521 tagccattga gatgttagat aggcaccata ctcacttttg ccctttagaa ggggaaagct
     2581 ggcaagattt tttacgtaat aacgctaaaa gttttagatg tgctttacta agtcatcgcg
     2641 atggagcaaa agtacattta ggtacacggc ctacagaaaa acagtatgaa actctcgaaa
     2701 atcaattagc ctttttatgc caacaaggtt tttcactaga gaatgcatta tatgcactca
     2761 gcgcagtggg gcattttact ttaggttgcg tattggaaga tcaagagcat caagtcgcta
     2821 aagaagaaag ggaaacacct actactgata gtatgccgcc attattacga caagctatcg
     2881 aattatttga tcaccaaggt gcagagccag ccttcttatt cggccttgaa ttgatcatat
     2941 gcggattaga aaaacaactt aaatgtgaaa gtgggtctta aggcatcaaa taaaacgaaa
     3001 ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcct
     3061 gagtaggaca aatccgccgc cctagaccta gggcgttcgg ctgcggcgag cggtatcagc
     3121 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcat gcaagaacat
     3181 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     3241 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     3301 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     3361 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     3421 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     3481 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     3541 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     3601 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     3661 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     3721 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     3781 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     3841 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     3901 gactagtgct tggattctca ccaataaaaa acgcccggcg gcaaccgagc gttctgaaca
     3961 aatccagatg gagttctgag gtcattactg gatctatcaa caggagtcca agcgagctcg
     4021 atatcaaatt acgccccgcc ctgccactca tcgcagtact gttgtaattc attaagcatt
     4081 ctgccgacat ggaagccatc acaaacggca tgatgaacct gaatcgccag cggcatcagc
     4141 accttgtcgc cttgcgtata atatttgccc atggtgaaaa cgggggcgaa gaagttgtcc
     4201 atattggcca cgtttaaatc aaaactggtg aaactcaccc agggattggc tgagacgaaa
     4261 aacatattct caataaaccc tttagggaaa taggccaggt tttcaccata acacgccaca
     4321 tcttgcgaat atatgtgtag aaactgccgg aaatcgtcgt ggtattcact ccagagcgat
     4381 gaaaacgttt cagtttgctc atggaaaacg gtgtaacaag ggtgaacact atcccatatc
     4441 accagctcac cgtctttcat tgccatacga aattccggat gagcattcat caggcgggca
     4501 agaatgtgaa taaaggccgg ataaaacttg tgcttatttt tctttacggt ctttaaaaag
     4561 gccgtaatat ccagctgaac ggtctggtta taggtacatt gagcaactga ctgaaatgcc
     4621 tcaaaatgtt ctttacgatg ccattgggat atatcaacgg tggtatatcc agtgattttt
     4681 ttctccattt tagcttcctt agctcctgaa aatctcgata actcaaaaaa tacgaccggt
     4741 agtgatctta tttcattatg gtgaaagttg gaacctctta cgggccggcc aacgtctcat
     4801 tttcgccaga tatcgacgtc ggagacctaa tgcgccagat
//
