LOCUS       sequence-245082-        4819 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of ompR(1-118aa)-ccaR(115-234aa) 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: pSR40.13
FEATURES             Location/Qualifiers
     source          1..4819
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     103..121
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(142..160,161..166,167..183,184..189,190..215)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    142..160
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    167..185
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      988..1059
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1075..1102
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             1398..1409
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1416..2129
                     /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      2156..2227
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2243..2270
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        2322..2356
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     CDS             2391..3017
                     /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(2467..2486)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      3023..3109
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(3274..3862)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3354..3373)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3950..4044)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(4065..4724)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4639..4658
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 gccggccaac gtctcatttt cgccagatat cgacgtcgga gacctaatgc gccagatatc
       61 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
      121 ccctttcgtc ttcacctcga gtccctatca gtgatagaga ttgacatccc tatcagtgat
      181 agagatactg agcacatcag caggacgcac tgaccgcata aaggacttag caaagcccaa
      241 ttttaaacaa atgcaagaga actacaagat tctggtggtc gatgacgaca tgcgcctgcg
      301 tgcgctgctg gaacgttatc tcaccgaaca aggcttccag gttcgaagcg tcgctaatgc
      361 agaacagatg gatcgcctgc tgactcgtga atctttccat cttatggtac tggatttaat
      421 gttacctggt gaagatggct tgtcgatttg ccgacgtctt cgtagtcaga gcaacccgat
      481 gccgatcatt atggtgacgg cgaaagggga agaagtggac cgtatcgtag gcctggagat
      541 tggcgctgac gactacattc caaaaccgtt taacccgcgt gaactgctgg cccgtatccg
      601 tgcgttgttg cgtcggggtt gtgcaacgtg ccaaccagtt ttagagtggg ggccaatcag
      661 gttggatcca agcacctatg aagttagtta tgacaatgag gttttgtctt tgacccgcaa
      721 ggaatacagc attctggaat tactactccg caatggccgt cgggtgctaa gtcggagcat
      781 gattatcgat agtatctgga agttggagag tcccccagag gaagatacgg ttaaggtgca
      841 tgtgcggagt ttgcgacaaa aattaaaaag tgccggttta tcagcagatg ccattgaaac
      901 ggtccatggc attgggtatc gtctggccaa tttaacggaa aaatctttgt gccaagggaa
      961 aaactagtaa taatctagac caggcatcaa ataaaacgaa aggctcagtc gaaagactgg
     1021 gcctttcgtt ttatctgttg tttgtcggtg aacgctctct actagagtca cactggctca
     1081 ccttcgggtg ggcctttctg cgtttatagt aaatcactgc ataattcgtg tcgctcaagg
     1141 cgcactcccg ttctggataa tgttttttgc gccgacatca taacggttct ggcaaatatt
     1201 ctgaaatgag ctggttagct agtcaagccc attgtgcttt tctctatcaa cctcagctta
     1261 cctgaagggg tgaacaggtc tgggttaatt catgttgcga aatgtaacag ttttagtcgc
     1321 atcagctaac tttccgattt ctttacgatt ttctccccct tttcttcaat tttactttgt
     1381 taggatcgca tttttaaaaa gaggagaaat actagatgcg taaaggcgaa gagctgttca
     1441 ctggtgtcgt ccctattctg gtggaactgg atggtgatgt caacggtcat aagttttccg
     1501 tgcgtggcga gggtgaaggt gacgcaacta atggtaaact gacgctgaag ttcatctgta
     1561 ctactggtaa actgccggta ccttggccga ctctggtaac gacgctgact tatggtgttc
     1621 agtgctttgc tcgttatccg gaccatatga agcagcatga cttcttcaag tccgccatgc
     1681 cggaaggcta tgtgcaggaa cgcacgattt cctttaagga tgacggcacg tacaaaacgc
     1741 gtgcggaagt gaaatttgaa ggcgataccc tggtaaaccg cattgagctg aaaggcattg
     1801 actttaaaga agacggcaat atcctgggcc ataagctgga atacaatttt aacagccaca
     1861 atgtttacat caccgccgat aaacaaaaaa atggcattaa agcgaatttt aaaattcgcc
     1921 acaacgtgga ggatggcagc gtgcagctgg ctgatcacta ccagcaaaac actccaatcg
     1981 gtgatggtcc tgttctgctg ccagacaatc actatctgag cacgcaaagc gttctgtcta
     2041 aagatccgaa cgagaaacgc gatcatatgg ttctgctgga gttcgtaacc gcagcgggca
     2101 tcacgcatgg tatggatgaa ctgtacaaat gatgataata atctagacca ggcatcaaat
     2161 aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa
     2221 cgctctctac tagagtcaca ctggctcacc ttcgggtggg cctttctgcg tttataggta
     2281 cccgtggatc ctctagagtc gacctgcagg catgcaagct ttttacggct agctcagtcc
     2341 taggtatagt gctagcccag ccagagaaac actctttaac agggggtagt atgatgtctc
     2401 gtttagataa aagtaaagtg attaacagcg cattagagct gcttaatgag gtcggaatcg
     2461 aaggtttaac aacccgtaaa ctcgcccaga agctaggtgt agagcagcct acattgtatt
     2521 ggcatgtaaa aaataagcgg gctttgctcg acgccttagc cattgagatg ttagataggc
     2581 accatactca cttttgccct ttagaagggg aaagctggca agatttttta cgtaataacg
     2641 ctaaaagttt tagatgtgct ttactaagtc atcgcgatgg agcaaaagta catttaggta
     2701 cacggcctac agaaaaacag tatgaaactc tcgaaaatca attagccttt ttatgccaac
     2761 aaggtttttc actagagaat gcattatatg cactcagcgc agtggggcat tttactttag
     2821 gttgcgtatt ggaagatcaa gagcatcaag tcgctaaaga agaaagggaa acacctacta
     2881 ctgatagtat gccgccatta ttacgacaag ctatcgaatt atttgatcac caaggtgcag
     2941 agccagcctt cttattcggc cttgaattga tcatatgcgg attagaaaaa caacttaaat
     3001 gtgaaagtgg gtcttaaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt
     3061 tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgcccta
     3121 gacctagggc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     3181 atccacagaa tcaggggata acgcatgcaa gaacatgtga gcaaaaggcc agcaaaaggc
     3241 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     3301 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     3361 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     3421 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     3481 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     3541 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     3601 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     3661 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
     3721 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     3781 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     3841 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     3901 gtggaacgaa aactcacgtt aagggatttt ggtcatgact agtgcttgga ttctcaccaa
     3961 taaaaaacgc ccggcggcaa ccgagcgttc tgaacaaatc cagatggagt tctgaggtca
     4021 ttactggatc tatcaacagg agtccaagcg agctcgatat caaattacgc cccgccctgc
     4081 cactcatcgc agtactgttg taattcatta agcattctgc cgacatggaa gccatcacaa
     4141 acggcatgat gaacctgaat cgccagcggc atcagcacct tgtcgccttg cgtataatat
     4201 ttgcccatgg tgaaaacggg ggcgaagaag ttgtccatat tggccacgtt taaatcaaaa
     4261 ctggtgaaac tcacccaggg attggctgag acgaaaaaca tattctcaat aaacccttta
     4321 gggaaatagg ccaggttttc accataacac gccacatctt gcgaatatat gtgtagaaac
     4381 tgccggaaat cgtcgtggta ttcactccag agcgatgaaa acgtttcagt ttgctcatgg
     4441 aaaacggtgt aacaagggtg aacactatcc catatcacca gctcaccgtc tttcattgcc
     4501 atacgaaatt ccggatgagc attcatcagg cgggcaagaa tgtgaataaa ggccggataa
     4561 aacttgtgct tatttttctt tacggtcttt aaaaaggccg taatatccag ctgaacggtc
     4621 tggttatagg tacattgagc aactgactga aatgcctcaa aatgttcttt acgatgccat
     4681 tgggatatat caacggtggt atatccagtg atttttttct ccattttagc ttccttagct
     4741 cctgaaaatc tcgataactc aaaaaatacg accggtagtg atcttatttc attatggtga
     4801 aagttggaac ctcttacgg
//
