LOCUS       sequence-245435-        4840 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of ompR(1-138aa)-ccaR(128-234aa) 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: pSR40.28
FEATURES             Location/Qualifiers
     source          1..4840
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     93..111
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(132..150,151..156,157..173,174..179,180..205)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    132..150
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    157..175
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      999..1070
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1086..1113
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             1409..1420
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1427..2140
                     /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      2167..2238
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2254..2281
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        2333..2367
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     CDS             2402..3028
                     /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(2478..2497)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      3034..3120
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(3285..3873)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3365..3384)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3961..4055)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(4076..4735)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4650..4669
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 gtctcatttt cgccagatat cgacgtcgga gacctaatgc gccagatatc gacgtctaag
       61 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc
      121 ttcacctcga gtccctatca gtgatagaga ttgacatccc tatcagtgat agagatactg
      181 agcacatcag caggacgcac tgaccgcata aaggacttag caaagcccaa ttttaaacaa
      241 atgcaagaga actacaagat tctggtggtc gatgacgaca tgcgcctgcg tgcgctgctg
      301 gaacgttatc tcaccgaaca aggcttccag gttcgaagcg tcgctaatgc agaacagatg
      361 gatcgcctgc tgactcgtga atctttccat cttatggtac tggatttaat gttacctggt
      421 gaagatggct tgtcgatttg ccgacgtctt cgtagtcaga gcaacccgat gccgatcatt
      481 atggtgacgg cgaaagggga agaagtggac cgtatcgtag gcctggagat tggcgctgac
      541 gactacattc caaaaccgtt taacccgcgt gaactgctgg cccgtatccg tgcggtgctg
      601 cgtcgtcagg cgaacgaact gccaggcgca ccgtcacagg aagaggcggt aattgagtgg
      661 gggccaatca ggttggatcc aagcacctat gaagttagtt atgacaatga ggttttgtct
      721 ttgacccgca aggaatacag cattctggaa ttactactcc gcaatggccg tcgggtgcta
      781 agtcggagca tgattatcga tagtatctgg aagttggaga gtcccccaga ggaagatacg
      841 gttaaggtgc atgtgcggag tttgcgacaa aaattaaaaa gtgccggttt atcagcagat
      901 gccattgaaa cggtccatgg cattgggtat cgtctggcca atttaacgga aaaatctttg
      961 tgccaaggga aaaactagta ataatctaga ccaggcatca aataaaacga aaggctcagt
     1021 cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc tactagagtc
     1081 acactggctc accttcgggt gggcctttct gcgtttatag taaatcactg cataattcgt
     1141 gtcgctcaag gcgcactccc gttctggata atgttttttg cgccgacatc ataacggttc
     1201 tggcaaatat tctgaaatga gctggttagc tagtcaagcc cattgtgctt ttctctatca
     1261 acctcagctt acctgaaggg gtgaacaggt ctgggttaat tcatgttgcg aaatgtaaca
     1321 gttttagtcg catcagctaa ctttccgatt tctttacgat tttctccccc ttttcttcaa
     1381 ttttactttg ttaggatcgc atttttaaaa agaggagaaa tactagatgc gtaaaggcga
     1441 agagctgttc actggtgtcg tccctattct ggtggaactg gatggtgatg tcaacggtca
     1501 taagttttcc gtgcgtggcg agggtgaagg tgacgcaact aatggtaaac tgacgctgaa
     1561 gttcatctgt actactggta aactgccggt accttggccg actctggtaa cgacgctgac
     1621 ttatggtgtt cagtgctttg ctcgttatcc ggaccatatg aagcagcatg acttcttcaa
     1681 gtccgccatg ccggaaggct atgtgcagga acgcacgatt tcctttaagg atgacggcac
     1741 gtacaaaacg cgtgcggaag tgaaatttga aggcgatacc ctggtaaacc gcattgagct
     1801 gaaaggcatt gactttaaag aagacggcaa tatcctgggc cataagctgg aatacaattt
     1861 taacagccac aatgtttaca tcaccgccga taaacaaaaa aatggcatta aagcgaattt
     1921 taaaattcgc cacaacgtgg aggatggcag cgtgcagctg gctgatcact accagcaaaa
     1981 cactccaatc ggtgatggtc ctgttctgct gccagacaat cactatctga gcacgcaaag
     2041 cgttctgtct aaagatccga acgagaaacg cgatcatatg gttctgctgg agttcgtaac
     2101 cgcagcgggc atcacgcatg gtatggatga actgtacaaa tgatgataat aatctagacc
     2161 aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt
     2221 ttgtcggtga acgctctcta ctagagtcac actggctcac cttcgggtgg gcctttctgc
     2281 gtttataggt acccgtggat cctctagagt cgacctgcag gcatgcaagc tttttacggc
     2341 tagctcagtc ctaggtatag tgctagccca gccagagaaa cactctttaa cagggggtag
     2401 tatgatgtct cgtttagata aaagtaaagt gattaacagc gcattagagc tgcttaatga
     2461 ggtcggaatc gaaggtttaa caacccgtaa actcgcccag aagctaggtg tagagcagcc
     2521 tacattgtat tggcatgtaa aaaataagcg ggctttgctc gacgccttag ccattgagat
     2581 gttagatagg caccatactc acttttgccc tttagaaggg gaaagctggc aagatttttt
     2641 acgtaataac gctaaaagtt ttagatgtgc tttactaagt catcgcgatg gagcaaaagt
     2701 acatttaggt acacggccta cagaaaaaca gtatgaaact ctcgaaaatc aattagcctt
     2761 tttatgccaa caaggttttt cactagagaa tgcattatat gcactcagcg cagtggggca
     2821 ttttacttta ggttgcgtat tggaagatca agagcatcaa gtcgctaaag aagaaaggga
     2881 aacacctact actgatagta tgccgccatt attacgacaa gctatcgaat tatttgatca
     2941 ccaaggtgca gagccagcct tcttattcgg ccttgaattg atcatatgcg gattagaaaa
     3001 acaacttaaa tgtgaaagtg ggtcttaagg catcaaataa aacgaaaggc tcagtcgaaa
     3061 gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag taggacaaat
     3121 ccgccgccct agacctaggg cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     3181 gtaatacggt tatccacaga atcaggggat aacgcatgca agaacatgtg agcaaaaggc
     3241 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     3301 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     3361 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     3421 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     3481 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     3541 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     3601 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     3661 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     3721 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     3781 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     3841 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     3901 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgac tagtgcttgg
     3961 attctcacca ataaaaaacg cccggcggca accgagcgtt ctgaacaaat ccagatggag
     4021 ttctgaggtc attactggat ctatcaacag gagtccaagc gagctcgata tcaaattacg
     4081 ccccgccctg ccactcatcg cagtactgtt gtaattcatt aagcattctg ccgacatgga
     4141 agccatcaca aacggcatga tgaacctgaa tcgccagcgg catcagcacc ttgtcgcctt
     4201 gcgtataata tttgcccatg gtgaaaacgg gggcgaagaa gttgtccata ttggccacgt
     4261 ttaaatcaaa actggtgaaa ctcacccagg gattggctga gacgaaaaac atattctcaa
     4321 taaacccttt agggaaatag gccaggtttt caccataaca cgccacatct tgcgaatata
     4381 tgtgtagaaa ctgccggaaa tcgtcgtggt attcactcca gagcgatgaa aacgtttcag
     4441 tttgctcatg gaaaacggtg taacaagggt gaacactatc ccatatcacc agctcaccgt
     4501 ctttcattgc catacgaaat tccggatgag cattcatcag gcgggcaaga atgtgaataa
     4561 aggccggata aaacttgtgc ttatttttct ttacggtctt taaaaaggcc gtaatatcca
     4621 gctgaacggt ctggttatag gtacattgag caactgactg aaatgcctca aaatgttctt
     4681 tacgatgcca ttgggatata tcaacggtgg tatatccagt gatttttttc tccattttag
     4741 cttccttagc tcctgaaaat ctcgataact caaaaaatac gaccggtagt gatcttattt
     4801 cattatggtg aaagttggaa cctcttacgg gccggccaac
//
