LOCUS       sequence-247804-        4819 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of ompR(1-110aa)-ccaR(107-234aa) chimera under PLtetO-1, 
            output Promoter PcpcG2-172.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4819)
  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 4819)
  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: pSR40.25
FEATURES             Location/Qualifiers
     source          1..4819
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     47..65
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(86..104,105..110,111..127,128..133,134..159)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    86..104
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    111..129
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      932..1003
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1019..1046
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             1342..1353
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1360..2073
                     /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      2100..2171
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2187..2214
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        2266..2300
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     CDS             2335..2961
                     /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(2411..2430)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      2967..3053
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(3218..3806)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3298..3317)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3894..3988)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(4009..4668)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LCSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4583..4602
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 tatcgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
       61 gaggcccttt cgtcttcacc tcgagtccct atcagtgata gagattgaca tccctatcag
      121 tgatagagat actgagcaca tcagcaggac gcactgaccg cataaaggac ttagcaaagc
      181 ccaattttaa acaaatgcaa gagaactaca agattctggt ggtcgatgac gacatgcgcc
      241 tgcgtgcgct gctggaacgt tatctcaccg aacaaggctt ccaggttcga agcgtcgcta
      301 atgcagaaca gatggatcgc ctgctgactc gtgaatcttt ccatcttatg gtactggatt
      361 taatgttacc tggtgaagat ggcttgtcga tttgccgacg tcttcgtagt cagagcaacc
      421 cgatgccgat cattatggtg acggcgaaag gggaagaagt ggaccgtatc gtaggcctgg
      481 agattggcgc tgacgactac attccaaaac cgtttaaccc gcgtgagtta tttgccaggg
      541 tgcgagcttt gttgcgtcgg ggttgtgcaa cgtgccaacc agttttagag tgggggccaa
      601 tcaggttgga tccaagcacc tatgaagtta gttatgacaa tgaggttttg tctttgaccc
      661 gcaaggaata cagcattctg gaattactac tccgcaatgg ccgtcgggtg ctaagtcgga
      721 gcatgattat cgatagtatc tggaagttgg agagtccccc agaggaagat acggttaagg
      781 tgcatgtgcg gagtttgcga caaaaattaa aaagtgccgg tttatcagca gatgccattg
      841 aaacggtcca tggcattggg tatcgtctgg ccaatttaac ggaaaaatct ttgtgccaag
      901 ggaaaaacta gtaataatct agaccaggca tcaaataaaa cgaaaggctc agtcgaaaga
      961 ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctctactaga gtcacactgg
     1021 ctcaccttcg ggtgggcctt tctgcgttta tagtaaatca ctgcataatt cgtgtcgctc
     1081 aaggcgcact cccgttctgg ataatgtttt ttgcgccgac atcataacgg ttctggcaaa
     1141 tattctgaaa tgagctggtt agctagtcaa gcccattgtg cttttctcta tcaacctcag
     1201 cttacctgaa ggggtgaaca ggtctgggtt aattcatgtt gcgaaatgta acagttttag
     1261 tcgcatcagc taactttccg atttctttac gattttctcc cccttttctt caattttact
     1321 ttgttaggat cgcattttta aaaagaggag aaatactaga tgcgtaaagg cgaagagctg
     1381 ttcactggtg tcgtccctat tctggtggaa ctggatggtg atgtcaacgg tcataagttt
     1441 tccgtgcgtg gcgagggtga aggtgacgca actaatggta aactgacgct gaagttcatc
     1501 tgtactactg gtaaactgcc ggtaccttgg ccgactctgg taacgacgct gacttatggt
     1561 gttcagtgct ttgctcgtta tccggaccat atgaagcagc atgacttctt caagtccgcc
     1621 atgccggaag gctatgtgca ggaacgcacg atttccttta aggatgacgg cacgtacaaa
     1681 acgcgtgcgg aagtgaaatt tgaaggcgat accctggtaa accgcattga gctgaaaggc
     1741 attgacttta aagaagacgg caatatcctg ggccataagc tggaatacaa ttttaacagc
     1801 cacaatgttt acatcaccgc cgataaacaa aaaaatggca ttaaagcgaa ttttaaaatt
     1861 cgccacaacg tggaggatgg cagcgtgcag ctggctgatc actaccagca aaacactcca
     1921 atcggtgatg gtcctgttct gctgccagac aatcactatc tgagcacgca aagcgttctg
     1981 tctaaagatc cgaacgagaa acgcgatcat atggttctgc tggagttcgt aaccgcagcg
     2041 ggcatcacgc atggtatgga tgaactgtac aaatgatgat aataatctag accaggcatc
     2101 aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg
     2161 tgaacgctct ctactagagt cacactggct caccttcggg tgggcctttc tgcgtttata
     2221 ggtacccgtg gatcctctag agtcgacctg caggcatgca agctttttac ggctagctca
     2281 gtcctaggta tagtgctagc ccagccagag aaacactctt taacaggggg tagtatgatg
     2341 tctcgtttag ataaaagtaa agtgattaac agcgcattag agctgcttaa tgaggtcgga
     2401 atcgaaggtt taacaacccg taaactcgcc cagaagctag gtgtagagca gcctacattg
     2461 tattggcatg taaaaaataa gcgggctttg ctcgacgcct tagccattga gatgttagat
     2521 aggcaccata ctcacttttg ccctttagaa ggggaaagct ggcaagattt tttacgtaat
     2581 aacgctaaaa gttttagatg tgctttacta agtcatcgcg atggagcaaa agtacattta
     2641 ggtacacggc ctacagaaaa acagtatgaa actctcgaaa atcaattagc ctttttatgc
     2701 caacaaggtt tttcactaga gaatgcatta tatgcactca gcgcagtggg gcattttact
     2761 ttaggttgcg tattggaaga tcaagagcat caagtcgcta aagaagaaag ggaaacacct
     2821 actactgata gtatgccgcc attattacga caagctatcg aattatttga tcaccaaggt
     2881 gcagagccag ccttcttatt cggccttgaa ttgatcatat gcggattaga aaaacaactt
     2941 aaatgtgaaa gtgggtctta aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg
     3001 cctttcgttt tatctgttgt ttgtcggtga acgctctcct gagtaggaca aatccgccgc
     3061 cctagaccta gggcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     3121 ggttatccac agaatcaggg gataacgcat gcaagaacat gtgagcaaaa ggccagcaaa
     3181 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     3241 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     3301 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     3361 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     3421 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     3481 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     3541 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     3601 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
     3661 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     3721 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     3781 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     3841 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gactagtgct tggattctca
     3901 ccaataaaaa acgcccggcg gcaaccgagc gttctgaaca aatccagatg gagttctgag
     3961 gtcattactg gatctatcaa caggagtcca agcgagctcg atatcaaatt acgccccgcc
     4021 ctgccactca tcgcagtact gttgtaattc attaagcatt ctgccgacat ggaagccatc
     4081 acaaacggca tgatgaacct gaatcgccag cggcatcagc accttgtcgc cttgcgtata
     4141 atatttgccc atggtgaaaa cgggggcgaa gaagttgtcc atattggcca cgtttaaatc
     4201 aaaactggtg aaactcaccc agggattggc tgagacgaaa aacatattct caataaaccc
     4261 tttagggaaa taggccaggt tttcaccata acacgccaca tcttgcgaat atatgtgtag
     4321 aaactgccgg aaatcgtcgt ggtattcact acagagcgat gaaaacgttt cagtttgctc
     4381 atggaaaacg gtgtaacaag ggtgaacact atcccatatc accagctcac cgtctttcat
     4441 tgccatacga aattccggat gagcattcat caggcgggca agaatgtgaa taaaggccgg
     4501 ataaaacttg tgcttatttt tctttacggt ctttaaaaag gccgtaatat ccagctgaac
     4561 ggtctggtta taggtacatt gagcaactga ctgaaatgcc tcaaaatgtt ctttacgatg
     4621 ccattgggat atatcaacgg tggtatatcc agtgattttt ttctccattt tagcttcctt
     4681 agctcctgaa aatctcgata actcaaaaaa tacgaccggt agtgatctta tttcattatg
     4741 gtgaaagttg gaacctctta cgggccggcc aacgtctcat tttcgccaga tatcgacgtc
     4801 ggagacctaa tgcgccaga
//
