LOCUS       sequence-245192-        4840 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of ompR(1-139aa)-ccaR(129-234aa) 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: pSR40.29
FEATURES             Location/Qualifiers
     source          1..4840
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     22..40
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(61..79,80..85,86..102,103..108,109..134)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    61..79
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    86..104
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      928..999
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1015..1042
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             1338..1349
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1356..2069
                     /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      2096..2167
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2183..2210
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        2262..2296
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     CDS             2331..2957
                     /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(2407..2426)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      2963..3049
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(3214..3802)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3294..3313)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3890..3984)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(4005..4664)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4579..4598
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct tcacctcgag
       61 tccctatcag tgatagagat tgacatccct atcagtgata gagatactga gcacatcagc
      121 aggacgcact gaccgcataa aggacttagc aaagcccaat tttaaacaaa tgcaagagaa
      181 ctacaagatt ctggtggtcg atgacgacat gcgcctgcgt gcgctgctgg aacgttatct
      241 caccgaacaa ggcttccagg ttcgaagcgt cgctaatgca gaacagatgg atcgcctgct
      301 gactcgtgaa tctttccatc ttatggtact ggatttaatg ttacctggtg aagatggctt
      361 gtcgatttgc cgacgtcttc gtagtcagag caacccgatg ccgatcatta tggtgacggc
      421 gaaaggggaa gaagtggacc gtatcgtagg cctggagatt ggcgctgacg actacattcc
      481 aaaaccgttt aacccgcgtg aactgctggc ccgtatccgt gcggtgctgc gtcgtcaggc
      541 gaacgaactg ccaggcgcac cgtcacagga agaggcggta attgcttggg ggccaatcag
      601 gttggatcca agcacctatg aagttagtta tgacaatgag gttttgtctt tgacccgcaa
      661 ggaatacagc attctggaat tactactccg caatggccgt cgggtgctaa gtcggagcat
      721 gattatcgat agtatctgga agttggagag tcccccagag gaagatacgg ttaaggtgca
      781 tgtgcggagt ttgcgacaaa aattaaaaag tgccggttta tcagcagatg ccattgaaac
      841 ggtccatggc attgggtatc gtctggccaa tttaacggaa aaatctttgt gccaagggaa
      901 aaactagtaa taatctagac caggcatcaa ataaaacgaa aggctcagtc gaaagactgg
      961 gcctttcgtt ttatctgttg tttgtcggtg aacgctctct actagagtca cactggctca
     1021 ccttcgggtg ggcctttctg cgtttatagt aaatcactgc ataattcgtg tcgctcaagg
     1081 cgcactcccg ttctggataa tgttttttgc gccgacatca taacggttct ggcaaatatt
     1141 ctgaaatgag ctggttagct agtcaagccc attgtgcttt tctctatcaa cctcagctta
     1201 cctgaagggg tgaacaggtc tgggttaatt catgttgcga aatgtaacag ttttagtcgc
     1261 atcagctaac tttccgattt ctttacgatt ttctccccct tttcttcaat tttactttgt
     1321 taggatcgca tttttaaaaa gaggagaaat actagatgcg taaaggcgaa gagctgttca
     1381 ctggtgtcgt ccctattctg gtggaactgg atggtgatgt caacggtcat aagttttccg
     1441 tgcgtggcga gggtgaaggt gacgcaacta atggtaaact gacgctgaag ttcatctgta
     1501 ctactggtaa actgccggta ccttggccga ctctggtaac gacgctgact tatggtgttc
     1561 agtgctttgc tcgttatccg gaccatatga agcagcatga cttcttcaag tccgccatgc
     1621 cggaaggcta tgtgcaggaa cgcacgattt cctttaagga tgacggcacg tacaaaacgc
     1681 gtgcggaagt gaaatttgaa ggcgataccc tggtaaaccg cattgagctg aaaggcattg
     1741 actttaaaga agacggcaat atcctgggcc ataagctgga atacaatttt aacagccaca
     1801 atgtttacat caccgccgat aaacaaaaaa atggcattaa agcgaatttt aaaattcgcc
     1861 acaacgtgga ggatggcagc gtgcagctgg ctgatcacta ccagcaaaac actccaatcg
     1921 gtgatggtcc tgttctgctg ccagacaatc actatctgag cacgcaaagc gttctgtcta
     1981 aagatccgaa cgagaaacgc gatcatatgg ttctgctgga gttcgtaacc gcagcgggca
     2041 tcacgcatgg tatggatgaa ctgtacaaat gatgataata atctagacca ggcatcaaat
     2101 aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa
     2161 cgctctctac tagagtcaca ctggctcacc ttcgggtggg cctttctgcg tttataggta
     2221 cccgtggatc ctctagagtc gacctgcagg catgcaagct ttttacggct agctcagtcc
     2281 taggtatagt gctagcccag ccagagaaac actctttaac agggggtagt atgatgtctc
     2341 gtttagataa aagtaaagtg attaacagcg cattagagct gcttaatgag gtcggaatcg
     2401 aaggtttaac aacccgtaaa ctcgcccaga agctaggtgt agagcagcct acattgtatt
     2461 ggcatgtaaa aaataagcgg gctttgctcg acgccttagc cattgagatg ttagataggc
     2521 accatactca cttttgccct ttagaagggg aaagctggca agatttttta cgtaataacg
     2581 ctaaaagttt tagatgtgct ttactaagtc atcgcgatgg agcaaaagta catttaggta
     2641 cacggcctac agaaaaacag tatgaaactc tcgaaaatca attagccttt ttatgccaac
     2701 aaggtttttc actagagaat gcattatatg cactcagcgc agtggggcat tttactttag
     2761 gttgcgtatt ggaagatcaa gagcatcaag tcgctaaaga agaaagggaa acacctacta
     2821 ctgatagtat gccgccatta ttacgacaag ctatcgaatt atttgatcac caaggtgcag
     2881 agccagcctt cttattcggc cttgaattga tcatatgcgg attagaaaaa caacttaaat
     2941 gtgaaagtgg gtcttaaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt
     3001 tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgcccta
     3061 gacctagggc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     3121 atccacagaa tcaggggata acgcatgcaa gaacatgtga gcaaaaggcc agcaaaaggc
     3181 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     3241 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     3301 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     3361 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     3421 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     3481 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     3541 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     3601 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
     3661 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     3721 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     3781 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     3841 gtggaacgaa aactcacgtt aagggatttt ggtcatgact agtgcttgga ttctcaccaa
     3901 taaaaaacgc ccggcggcaa ccgagcgttc tgaacaaatc cagatggagt tctgaggtca
     3961 ttactggatc tatcaacagg agtccaagcg agctcgatat caaattacgc cccgccctgc
     4021 cactcatcgc agtactgttg taattcatta agcattctgc cgacatggaa gccatcacaa
     4081 acggcatgat gaacctgaat cgccagcggc atcagcacct tgtcgccttg cgtataatat
     4141 ttgcccatgg tgaaaacggg ggcgaagaag ttgtccatat tggccacgtt taaatcaaaa
     4201 ctggtgaaac tcacccaggg attggctgag acgaaaaaca tattctcaat aaacccttta
     4261 gggaaatagg ccaggttttc accataacac gccacatctt gcgaatatat gtgtagaaac
     4321 tgccggaaat cgtcgtggta ttcactccag agcgatgaaa acgtttcagt ttgctcatgg
     4381 aaaacggtgt aacaagggtg aacactatcc catatcacca gctcaccgtc tttcattgcc
     4441 atacgaaatt ccggatgagc attcatcagg cgggcaagaa tgtgaataaa ggccggataa
     4501 aacttgtgct tatttttctt tacggtcttt aaaaaggccg taatatccag ctgaacggtc
     4561 tggttatagg tacattgagc aactgactga aatgcctcaa aatgttcttt acgatgccat
     4621 tgggatatat caacggtggt atatccagtg atttttttct ccattttagc ttccttagct
     4681 cctgaaaatc tcgataactc aaaaaatacg accggtagtg atcttatttc attatggtga
     4741 aagttggaac ctcttacggg ccggccaacg tctcattttc gccagatatc gacgtcggag
     4801 acctaatgcg ccagatatcg acgtctaaga aaccattatt
//
