LOCUS       sequence-245085-        4819 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of ompR(1-122aa)-ccaR(119-234aa) 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: pSR40.14
FEATURES             Location/Qualifiers
     source          1..4819
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     113..131
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(152..170,171..176,177..193,194..199,200..225)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    152..170
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    177..195
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      998..1069
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1085..1112
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             1408..1419
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1426..2139
                     /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      2166..2237
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2253..2280
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        2332..2366
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     CDS             2401..3027
                     /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(2477..2496)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      3033..3119
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(3284..3872)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3364..3383)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3960..4054)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(4075..4734)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4649..4668
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 cctcttacgg gccggccaac gtctcatttt cgccagatat cgacgtcgga gacctaatgc
       61 gccagatatc gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
      121 tatcacgagg ccctttcgtc ttcacctcga gtccctatca gtgatagaga ttgacatccc
      181 tatcagtgat agagatactg agcacatcag caggacgcac tgaccgcata aaggacttag
      241 caaagcccaa ttttaaacaa atgcaagaga actacaagat tctggtggtc gatgacgaca
      301 tgcgcctgcg tgcgctgctg gaacgttatc tcaccgaaca aggcttccag gttcgaagcg
      361 tcgctaatgc agaacagatg gatcgcctgc tgactcgtga atctttccat cttatggtac
      421 tggatttaat gttacctggt gaagatggct tgtcgatttg ccgacgtctt cgtagtcaga
      481 gcaacccgat gccgatcatt atggtgacgg cgaaagggga agaagtggac cgtatcgtag
      541 gcctggagat tggcgctgac gactacattc caaaaccgtt taacccgcgt gaactgctgg
      601 cccgtatccg tgcggtgctg cgtcgtggtt gtgcaacgtg ccaaccagtt ttagagtggg
      661 ggccaatcag gttggatcca agcacctatg aagttagtta tgacaatgag gttttgtctt
      721 tgacccgcaa ggaatacagc attctggaat tactactccg caatggccgt cgggtgctaa
      781 gtcggagcat gattatcgat agtatctgga agttggagag tcccccagag gaagatacgg
      841 ttaaggtgca tgtgcggagt ttgcgacaaa aattaaaaag tgccggttta tcagcagatg
      901 ccattgaaac ggtccatggc attgggtatc gtctggccaa tttaacggaa aaatctttgt
      961 gccaagggaa aaactagtaa taatctagac caggcatcaa ataaaacgaa aggctcagtc
     1021 gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctct actagagtca
     1081 cactggctca ccttcgggtg ggcctttctg cgtttatagt aaatcactgc ataattcgtg
     1141 tcgctcaagg cgcactcccg ttctggataa tgttttttgc gccgacatca taacggttct
     1201 ggcaaatatt ctgaaatgag ctggttagct agtcaagccc attgtgcttt tctctatcaa
     1261 cctcagctta cctgaagggg tgaacaggtc tgggttaatt catgttgcga aatgtaacag
     1321 ttttagtcgc atcagctaac tttccgattt ctttacgatt ttctccccct tttcttcaat
     1381 tttactttgt taggatcgca tttttaaaaa gaggagaaat actagatgcg taaaggcgaa
     1441 gagctgttca ctggtgtcgt ccctattctg gtggaactgg atggtgatgt caacggtcat
     1501 aagttttccg tgcgtggcga gggtgaaggt gacgcaacta atggtaaact gacgctgaag
     1561 ttcatctgta ctactggtaa actgccggta ccttggccga ctctggtaac gacgctgact
     1621 tatggtgttc agtgctttgc tcgttatccg gaccatatga agcagcatga cttcttcaag
     1681 tccgccatgc cggaaggcta tgtgcaggaa cgcacgattt cctttaagga tgacggcacg
     1741 tacaaaacgc gtgcggaagt gaaatttgaa ggcgataccc tggtaaaccg cattgagctg
     1801 aaaggcattg actttaaaga agacggcaat atcctgggcc ataagctgga atacaatttt
     1861 aacagccaca atgtttacat caccgccgat aaacaaaaaa atggcattaa agcgaatttt
     1921 aaaattcgcc acaacgtgga ggatggcagc gtgcagctgg ctgatcacta ccagcaaaac
     1981 actccaatcg gtgatggtcc tgttctgctg ccagacaatc actatctgag cacgcaaagc
     2041 gttctgtcta aagatccgaa cgagaaacgc gatcatatgg ttctgctgga gttcgtaacc
     2101 gcagcgggca tcacgcatgg tatggatgaa ctgtacaaat gatgataata atctagacca
     2161 ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt
     2221 tgtcggtgaa cgctctctac tagagtcaca ctggctcacc ttcgggtggg cctttctgcg
     2281 tttataggta cccgtggatc ctctagagtc gacctgcagg catgcaagct ttttacggct
     2341 agctcagtcc taggtatagt gctagcccag ccagagaaac actctttaac agggggtagt
     2401 atgatgtctc gtttagataa aagtaaagtg attaacagcg cattagagct gcttaatgag
     2461 gtcggaatcg aaggtttaac aacccgtaaa ctcgcccaga agctaggtgt agagcagcct
     2521 acattgtatt ggcatgtaaa aaataagcgg gctttgctcg acgccttagc cattgagatg
     2581 ttagataggc accatactca cttttgccct ttagaagggg aaagctggca agatttttta
     2641 cgtaataacg ctaaaagttt tagatgtgct ttactaagtc atcgcgatgg agcaaaagta
     2701 catttaggta cacggcctac agaaaaacag tatgaaactc tcgaaaatca attagccttt
     2761 ttatgccaac aaggtttttc actagagaat gcattatatg cactcagcgc agtggggcat
     2821 tttactttag gttgcgtatt ggaagatcaa gagcatcaag tcgctaaaga agaaagggaa
     2881 acacctacta ctgatagtat gccgccatta ttacgacaag ctatcgaatt atttgatcac
     2941 caaggtgcag agccagcctt cttattcggc cttgaattga tcatatgcgg attagaaaaa
     3001 caacttaaat gtgaaagtgg gtcttaaggc atcaaataaa acgaaaggct cagtcgaaag
     3061 actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc
     3121 cgccgcccta gacctagggc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     3181 taatacggtt atccacagaa tcaggggata acgcatgcaa gaacatgtga gcaaaaggcc
     3241 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     3301 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     3361 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     3421 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     3481 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     3541 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     3601 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     3661 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     3721 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     3781 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     3841 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     3901 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgact agtgcttgga
     3961 ttctcaccaa taaaaaacgc ccggcggcaa ccgagcgttc tgaacaaatc cagatggagt
     4021 tctgaggtca ttactggatc tatcaacagg agtccaagcg agctcgatat caaattacgc
     4081 cccgccctgc cactcatcgc agtactgttg taattcatta agcattctgc cgacatggaa
     4141 gccatcacaa acggcatgat gaacctgaat cgccagcggc atcagcacct tgtcgccttg
     4201 cgtataatat ttgcccatgg tgaaaacggg ggcgaagaag ttgtccatat tggccacgtt
     4261 taaatcaaaa ctggtgaaac tcacccaggg attggctgag acgaaaaaca tattctcaat
     4321 aaacccttta gggaaatagg ccaggttttc accataacac gccacatctt gcgaatatat
     4381 gtgtagaaac tgccggaaat cgtcgtggta ttcactccag agcgatgaaa acgtttcagt
     4441 ttgctcatgg aaaacggtgt aacaagggtg aacactatcc catatcacca gctcaccgtc
     4501 tttcattgcc atacgaaatt ccggatgagc attcatcagg cgggcaagaa tgtgaataaa
     4561 ggccggataa aacttgtgct tatttttctt tacggtcttt aaaaaggccg taatatccag
     4621 ctgaacggtc tggttatagg tacattgagc aactgactga aatgcctcaa aatgttcttt
     4681 acgatgccat tgggatatat caacggtggt atatccagtg atttttttct ccattttagc
     4741 ttccttagct cctgaaaatc tcgataactc aaaaaatacg accggtagtg atcttatttc
     4801 attatggtga aagttggaa
//
