LOCUS       sequence-245434-        4819 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of ompR(1-113aa)-ccaR(110-234aa) 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: pSR40.26
FEATURES             Location/Qualifiers
     source          1..4819
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     24..42
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(63..81,82..87,88..104,105..110,111..136)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    63..81
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    88..106
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      909..980
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      996..1023
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             1319..1330
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1337..2050
                     /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      2077..2148
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2164..2191
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        2243..2277
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     CDS             2312..2938
                     /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(2388..2407)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      2944..3030
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(3195..3783)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3275..3294)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3871..3965)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3986..4645)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4560..4579
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt cttcacctcg
       61 agtccctatc agtgatagag attgacatcc ctatcagtga tagagatact gagcacatca
      121 gcaggacgca ctgaccgcat aaaggactta gcaaagccca attttaaaca aatgcaagag
      181 aactacaaga ttctggtggt cgatgacgac atgcgcctgc gtgcgctgtt ggaacgttat
      241 ctcaccgaac aaggcttcca ggttcgaagc gtcgctaatg cagaacagat ggatcgcctg
      301 ctgactcgtg aatctttcca tcttatggta ctggatttaa tgttacctgg tgaagatggc
      361 ttgtcgattt gccgacgtct tcgtagtcag agcaacccga tgccgatcat tatggtgacg
      421 gcgaaagggg aagaagtgga ccgtatcgta ggcctggaga ttggcgctga cgactacatt
      481 ccaaaaccgt ttaacccgcg tgaactgctg gccagggtgc gagctttgtt gcgtcggggt
      541 tgtgcaacgt gccaaccagt tttagagtgg gggccaatca ggttggatcc aagcacctat
      601 gaagttagtt atgacaatga ggttttgtct ttgacccgca aggaatacag cattctggaa
      661 ttactactcc gcaatggccg tcgggtgcta agtcggagca tgattatcga tagtatctgg
      721 aagttggaga gtcccccaga ggaagatacg gttaaggtgc atgtgcggag tttgcgacaa
      781 aaattaaaaa gtgccggttt atcagcagat gccattgaaa cggtccatgg cattgggtat
      841 cgtctggcca atttaacgga aaaatctttg tgccaaggga aaaactagta ataatctaga
      901 ccaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt
      961 gtttgtcggt gaacgctctc tactagagtc acactggctc accttcgggt gggcctttct
     1021 gcgtttatag taaatcactg cataattcgt gtcgctcaag gcgcactccc gttctggata
     1081 atgttttttg cgccgacatc ataacggttc tggcaaatat tctgaaatga gctggttagc
     1141 tagtcaagcc cattgtgctt ttctctatca acctcagctt acctgaaggg gtgaacaggt
     1201 ctgggttaat tcatgttgcg aaatgtaaca gttttagtcg catcagctaa ctttccgatt
     1261 tctttacgat tttctccccc ttttcttcaa ttttactttg ttaggatcgc atttttaaaa
     1321 agaggagaaa tactagatgc gtaaaggcga agagctgttc actggtgtcg tccctattct
     1381 ggtggaactg gatggtgatg tcaacggtca taagttttcc gtgcgtggcg agggtgaagg
     1441 tgacgcaact aatggtaaac tgacgctgaa gttcatctgt actactggta aactgccggt
     1501 accttggccg actctggtaa cgacgctgac ttatggtgtt cagtgctttg ctcgttatcc
     1561 ggaccatatg aagcagcatg acttcttcaa gtccgccatg ccggaaggct atgtgcagga
     1621 acgcacgatt tcctttaagg atgacggcac gtacaaaacg cgtgcggaag tgaaatttga
     1681 aggcgatacc ctggtaaacc gcattgagct gaaaggcatt gactttaaag aagacggcaa
     1741 tatcctgggc cataagctgg aatacaattt taacagccac aatgtttaca tcaccgccga
     1801 taaacaaaaa aatggcatta aagcgaattt taaaattcgc cacaacgtgg aggatggcag
     1861 cgtgcagctg gctgatcact accagcaaaa cactccaatc ggtgatggtc ctgttctgct
     1921 gccagacaat cactatctga gcacgcaaag cgttctgtct aaagatccga acgagaaacg
     1981 cgatcatatg gttctgctgg agttcgtaac cgcagcgggc atcacgcatg gtatggatga
     2041 actgtacaaa tgatgataat aatctagacc aggcatcaaa taaaacgaaa ggctcagtcg
     2101 aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcta ctagagtcac
     2161 actggctcac cttcgggtgg gcctttctgc gtttataggt acccgtggat cctctagagt
     2221 cgacctgcag gcatgcaagc tttttacggc tagctcagtc ctaggtatag tgctagccca
     2281 gccagagaaa cactctttaa cagggggtag tatgatgtct cgtttagata aaagtaaagt
     2341 gattaacagc gcattagagc tgcttaatga ggtcggaatc gaaggtttaa caacccgtaa
     2401 actcgcccag aagctaggtg tagagcagcc tacattgtat tggcatgtaa aaaataagcg
     2461 ggctttgctc gacgccttag ccattgagat gttagatagg caccatactc acttttgccc
     2521 tttagaaggg gaaagctggc aagatttttt acgtaataac gctaaaagtt ttagatgtgc
     2581 tttactaagt catcgcgatg gagcaaaagt acatttaggt acacggccta cagaaaaaca
     2641 gtatgaaact ctcgaaaatc aattagcctt tttatgccaa caaggttttt cactagagaa
     2701 tgcattatat gcactcagcg cagtggggca ttttacttta ggttgcgtat tggaagatca
     2761 agagcatcaa gtcgctaaag aagaaaggga aacacctact actgatagta tgccgccatt
     2821 attacgacaa gctatcgaat tatttgatca ccaaggtgca gagccagcct tcttattcgg
     2881 ccttgaattg atcatatgcg gattagaaaa acaacttaaa tgtgaaagtg ggtcttaagg
     2941 catcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg
     3001 tcggtgaacg ctctcctgag taggacaaat ccgccgccct agacctaggg cgttcggctg
     3061 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     3121 aacgcatgca agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     3181 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     3241 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     3301 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     3361 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     3421 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     3481 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     3541 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     3601 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
     3661 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     3721 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     3781 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     3841 taagggattt tggtcatgac tagtgcttgg attctcacca ataaaaaacg cccggcggca
     3901 accgagcgtt ctgaacaaat ccagatggag ttctgaggtc attactggat ctatcaacag
     3961 gagtccaagc gagctcgata tcaaattacg ccccgccctg ccactcatcg cagtactgtt
     4021 gtaattcatt aagcattctg ccgacatgga agccatcaca aacggcatga tgaacctgaa
     4081 tcgccagcgg catcagcacc ttgtcgcctt gcgtataata tttgcccatg gtgaaaacgg
     4141 gggcgaagaa gttgtccata ttggccacgt ttaaatcaaa actggtgaaa ctcacccagg
     4201 gattggctga gacgaaaaac atattctcaa taaacccttt agggaaatag gccaggtttt
     4261 caccataaca cgccacatct tgcgaatata tgtgtagaaa ctgccggaaa tcgtcgtggt
     4321 attcactcca gagcgatgaa aacgtttcag tttgctcatg gaaaacggtg taacaagggt
     4381 gaacactatc ccatatcacc agctcaccgt ctttcattgc catacgaaat tccggatgag
     4441 cattcatcag gcgggcaaga atgtgaataa aggccggata aaacttgtgc ttatttttct
     4501 ttacggtctt taaaaaggcc gtaatatcca gctgaacggt ctggttatag gtacattgag
     4561 caactgactg aaatgcctca aaatgttctt tacgatgcca ttgggatata tcaacggtgg
     4621 tatatccagt gatttttttc tccattttag cttccttagc tcctgaaaat ctcgataact
     4681 caaaaaatac gaccggtagt gatcttattt cattatggtg aaagttggaa cctcttacgg
     4741 gccggccaac gtctcatttt cgccagatat cgacgtcgga gacctaatgc gccagatatc
     4801 gacgtctaag aaaccatta
//
