LOCUS       sequence-245188-        4840 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of ompR(1-137aa)-ccaR(127-234aa) 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: pSR40.15
FEATURES             Location/Qualifiers
     source          1..4840
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     49..67
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(88..106,107..112,113..129,130..135,136..161)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    88..106
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    113..131
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      955..1026
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1042..1069
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             1365..1376
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1383..2096
                     /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      2123..2194
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2210..2237
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        2289..2323
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     CDS             2358..2984
                     /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(2434..2453)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      2990..3076
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(3241..3829)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3321..3340)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3917..4011)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(4032..4691)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4606..4625
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 gatatcgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
       61 acgaggccct ttcgtcttca cctcgagtcc ctatcagtga tagagattga catccctatc
      121 agtgatagag atactgagca catcagcagg acgcactgac cgcataaagg acttagcaaa
      181 gcccaatttt aaacaaatgc aagagaacta caagattctg gtggtcgatg acgacatgcg
      241 cctgcgtgcg ctgctggaac gttatctcac cgaacaaggc ttccaggttc gaagcgtcgc
      301 taatgcagaa cagatggatc gcctgctgac tcgtgaatct ttccatctta tggtactgga
      361 tttaatgtta cctggtgaag atggcttgtc gatttgccga cgtcttcgta gtcagagcaa
      421 cccgatgccg atcattatgg tgacggcgaa aggggaagaa gtggaccgta tcgtaggcct
      481 ggagattggc gctgacgact acattccaaa accgtttaac ccgcgtgaac tgctggcccg
      541 tatccgtgcg gtgctgcgtc gtcaggcgaa cgaactgcca ggcgcaccgt cacaggaaga
      601 ggcggtatta gagtgggggc caatcaggtt ggatccaagc acctatgaag ttagttatga
      661 caatgaggtt ttgtctttga cccgcaagga atacagcatt ctggaattac tactccgcaa
      721 tggccgtcgg gtgctaagtc ggagcatgat tatcgatagt atctggaagt tggagagtcc
      781 cccagaggaa gatacggtta aggtgcatgt gcggagtttg cgacaaaaat taaaaagtgc
      841 cggtttatca gcagatgcca ttgaaacggt ccatggcatt gggtatcgtc tggccaattt
      901 aacggaaaaa tctttgtgcc aagggaaaaa ctagtaataa tctagaccag gcatcaaata
      961 aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac
     1021 gctctctact agagtcacac tggctcacct tcgggtgggc ctttctgcgt ttatagtaaa
     1081 tcactgcata attcgtgtcg ctcaaggcgc actcccgttc tggataatgt tttttgcgcc
     1141 gacatcataa cggttctggc aaatattctg aaatgagctg gttagctagt caagcccatt
     1201 gtgcttttct ctatcaacct cagcttacct gaaggggtga acaggtctgg gttaattcat
     1261 gttgcgaaat gtaacagttt tagtcgcatc agctaacttt ccgatttctt tacgattttc
     1321 tccccctttt cttcaatttt actttgttag gatcgcattt ttaaaaagag gagaaatact
     1381 agatgcgtaa aggcgaagag ctgttcactg gtgtcgtccc tattctggtg gaactggatg
     1441 gtgatgtcaa cggtcataag ttttccgtgc gtggcgaggg tgaaggtgac gcaactaatg
     1501 gtaaactgac gctgaagttc atctgtacta ctggtaaact gccggtacct tggccgactc
     1561 tggtaacgac gctgacttat ggtgttcagt gctttgctcg ttatccggac catatgaagc
     1621 agcatgactt cttcaagtcc gccatgccgg aaggctatgt gcaggaacgc acgatttcct
     1681 ttaaggatga cggcacgtac aaaacgcgtg cggaagtgaa atttgaaggc gataccctgg
     1741 taaaccgcat tgagctgaaa ggcattgact ttaaagaaga cggcaatatc ctgggccata
     1801 agctggaata caattttaac agccacaatg tttacatcac cgccgataaa caaaaaaatg
     1861 gcattaaagc gaattttaaa attcgccaca acgtggagga tggcagcgtg cagctggctg
     1921 atcactacca gcaaaacact ccaatcggtg atggtcctgt tctgctgcca gacaatcact
     1981 atctgagcac gcaaagcgtt ctgtctaaag atccgaacga gaaacgcgat catatggttc
     2041 tgctggagtt cgtaaccgca gcgggcatca cgcatggtat ggatgaactg tacaaatgat
     2101 gataataatc tagaccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt
     2161 tcgttttatc tgttgtttgt cggtgaacgc tctctactag agtcacactg gctcaccttc
     2221 gggtgggcct ttctgcgttt ataggtaccc gtggatcctc tagagtcgac ctgcaggcat
     2281 gcaagctttt tacggctagc tcagtcctag gtatagtgct agcccagcca gagaaacact
     2341 ctttaacagg gggtagtatg atgtctcgtt tagataaaag taaagtgatt aacagcgcat
     2401 tagagctgct taatgaggtc ggaatcgaag gtttaacaac ccgtaaactc gcccagaagc
     2461 taggtgtaga gcagcctaca ttgtattggc atgtaaaaaa taagcgggct ttgctcgacg
     2521 ccttagccat tgagatgtta gataggcacc atactcactt ttgcccttta gaaggggaaa
     2581 gctggcaaga ttttttacgt aataacgcta aaagttttag atgtgcttta ctaagtcatc
     2641 gcgatggagc aaaagtacat ttaggtacac ggcctacaga aaaacagtat gaaactctcg
     2701 aaaatcaatt agccttttta tgccaacaag gtttttcact agagaatgca ttatatgcac
     2761 tcagcgcagt ggggcatttt actttaggtt gcgtattgga agatcaagag catcaagtcg
     2821 ctaaagaaga aagggaaaca cctactactg atagtatgcc gccattatta cgacaagcta
     2881 tcgaattatt tgatcaccaa ggtgcagagc cagccttctt attcggcctt gaattgatca
     2941 tatgcggatt agaaaaacaa cttaaatgtg aaagtgggtc ttaaggcatc aaataaaacg
     3001 aaaggctcag tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct
     3061 cctgagtagg acaaatccgc cgccctagac ctagggcgtt cggctgcggc gagcggtatc
     3121 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg catgcaagaa
     3181 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     3241 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     3301 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     3361 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     3421 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     3481 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     3541 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     3601 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     3661 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     3721 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     3781 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     3841 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     3901 catgactagt gcttggattc tcaccaataa aaaacgcccg gcggcaaccg agcgttctga
     3961 acaaatccag atggagttct gaggtcatta ctggatctat caacaggagt ccaagcgagc
     4021 tcgatatcaa attacgcccc gccctgccac tcatcgcagt actgttgtaa ttcattaagc
     4081 attctgccga catggaagcc atcacaaacg gcatgatgaa cctgaatcgc cagcggcatc
     4141 agcaccttgt cgccttgcgt ataatatttg cccatggtga aaacgggggc gaagaagttg
     4201 tccatattgg ccacgtttaa atcaaaactg gtgaaactca cccagggatt ggctgagacg
     4261 aaaaacatat tctcaataaa ccctttaggg aaataggcca ggttttcacc ataacacgcc
     4321 acatcttgcg aatatatgtg tagaaactgc cggaaatcgt cgtggtattc actccagagc
     4381 gatgaaaacg tttcagtttg ctcatggaaa acggtgtaac aagggtgaac actatcccat
     4441 atcaccagct caccgtcttt cattgccata cgaaattccg gatgagcatt catcaggcgg
     4501 gcaagaatgt gaataaaggc cggataaaac ttgtgcttat ttttctttac ggtctttaaa
     4561 aaggccgtaa tatccagctg aacggtctgg ttataggtac attgagcaac tgactgaaat
     4621 gcctcaaaat gttctttacg atgccattgg gatatatcaa cggtggtata tccagtgatt
     4681 tttttctcca ttttagcttc cttagctcct gaaaatctcg ataactcaaa aaatacgacc
     4741 ggtagtgatc ttatttcatt atggtgaaag ttggaacctc ttacgggccg gccaacgtct
     4801 cattttcgcc agatatcgac gtcggagacc taatgcgcca
//
