LOCUS       sequence-245190-        4840 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of ompR(1-150aa)-ccaR(140-234aa) 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: pSR40.18
FEATURES             Location/Qualifiers
     source          1..4840
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     147..165
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(186..204,205..210,211..227,228..233,234..259)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    186..204
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    211..229
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      1053..1124
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1140..1167
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             1463..1474
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1481..2194
                     /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      2221..2292
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2308..2335
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        2387..2421
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     CDS             2456..3082
                     /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(2532..2551)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      3088..3174
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(3339..3927)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3419..3438)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(4015..4109)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(4130..4789)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4704..4723
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 tagtgatctt atttcattat ggtgaaagtt ggaacctctt acgggccggc caacgtctca
       61 ttttcgccag atatcgacgt cggagaccta atgcgccaga tatcgacgtc taagaaacca
      121 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtcttcacc
      181 tcgagtccct atcagtgata gagattgaca tccctatcag tgatagagat actgagcaca
      241 tcagcaggac gcactgaccg cataaaggac ttagcaaagc ccaattttaa acaaatgcaa
      301 gagaactaca agattctggt ggtcgatgac gacatgcgcc tgcgtgcgct gctggaacgt
      361 tatctcaccg aacaaggctt ccaggttcga agcgtcgcta atgcagaaca gatggatcgc
      421 ctgctgactc gtgaatcttt ccatcttatg gtactggatt taatgttacc tggtgaagat
      481 ggcttgtcga tttgccgacg tcttcgtagt cagagcaacc cgatgccgat cattatggtg
      541 acggcgaaag gggaagaagt ggaccgtatc gtaggcctgg agattggcgc tgacgactac
      601 attccaaaac cgtttaaccc gcgtgaactg ctggcccgta tccgtgcggt gctgcgtcgt
      661 caggcgaacg aactgccagg cgcaccgtca caggaagagg cggtaattgc tttcggtaag
      721 ttcaaactta acctcggtac gcgcgaagtt agttatgaca atgaggtttt gtctttgacc
      781 cgcaaggaat acagcattct ggaattacta ctccgcaatg gccgtcgggt gctaagtcgg
      841 agcatgatta tcgatagtat ctggaagttg gagagtcccc cagaggaaga tacggttaag
      901 gtgcatgtgc ggagtttgcg acaaaaatta aaaagtgccg gtttatcagc agatgccatt
      961 gaaacggtcc atggcattgg gtatcgtctg gccaatttaa cggaaaaatc tttgtgccaa
     1021 gggaaaaact agtaataatc tagaccaggc atcaaataaa acgaaaggct cagtcgaaag
     1081 actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctctactag agtcacactg
     1141 gctcaccttc gggtgggcct ttctgcgttt atagtaaatc actgcataat tcgtgtcgct
     1201 caaggcgcac tcccgttctg gataatgttt tttgcgccga catcataacg gttctggcaa
     1261 atattctgaa atgagctggt tagctagtca agcccattgt gcttttctct atcaacctca
     1321 gcttacctga aggggtgaac aggtctgggt taattcatgt tgcgaaatgt aacagtttta
     1381 gtcgcatcag ctaactttcc gatttcttta cgattttctc ccccttttct tcaattttac
     1441 tttgttagga tcgcattttt aaaaagagga gaaatactag atgcgtaaag gcgaagagct
     1501 gttcactggt gtcgtcccta ttctggtgga actggatggt gatgtcaacg gtcataagtt
     1561 ttccgtgcgt ggcgagggtg aaggtgacgc aactaatggt aaactgacgc tgaagttcat
     1621 ctgtactact ggtaaactgc cggtaccttg gccgactctg gtaacgacgc tgacttatgg
     1681 tgttcagtgc tttgctcgtt atccggacca tatgaagcag catgacttct tcaagtccgc
     1741 catgccggaa ggctatgtgc aggaacgcac gatttccttt aaggatgacg gcacgtacaa
     1801 aacgcgtgcg gaagtgaaat ttgaaggcga taccctggta aaccgcattg agctgaaagg
     1861 cattgacttt aaagaagacg gcaatatcct gggccataag ctggaataca attttaacag
     1921 ccacaatgtt tacatcaccg ccgataaaca aaaaaatggc attaaagcga attttaaaat
     1981 tcgccacaac gtggaggatg gcagcgtgca gctggctgat cactaccagc aaaacactcc
     2041 aatcggtgat ggtcctgttc tgctgccaga caatcactat ctgagcacgc aaagcgttct
     2101 gtctaaagat ccgaacgaga aacgcgatca tatggttctg ctggagttcg taaccgcagc
     2161 gggcatcacg catggtatgg atgaactgta caaatgatga taataatcta gaccaggcat
     2221 caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg
     2281 gtgaacgctc tctactagag tcacactggc tcaccttcgg gtgggccttt ctgcgtttat
     2341 aggtacccgt ggatcctcta gagtcgacct gcaggcatgc aagcttttta cggctagctc
     2401 agtcctaggt atagtgctag cccagccaga gaaacactct ttaacagggg gtagtatgat
     2461 gtctcgttta gataaaagta aagtgattaa cagcgcatta gagctgctta atgaggtcgg
     2521 aatcgaaggt ttaacaaccc gtaaactcgc ccagaagcta ggtgtagagc agcctacatt
     2581 gtattggcat gtaaaaaata agcgggcttt gctcgacgcc ttagccattg agatgttaga
     2641 taggcaccat actcactttt gccctttaga aggggaaagc tggcaagatt ttttacgtaa
     2701 taacgctaaa agttttagat gtgctttact aagtcatcgc gatggagcaa aagtacattt
     2761 aggtacacgg cctacagaaa aacagtatga aactctcgaa aatcaattag cctttttatg
     2821 ccaacaaggt ttttcactag agaatgcatt atatgcactc agcgcagtgg ggcattttac
     2881 tttaggttgc gtattggaag atcaagagca tcaagtcgct aaagaagaaa gggaaacacc
     2941 tactactgat agtatgccgc cattattacg acaagctatc gaattatttg atcaccaagg
     3001 tgcagagcca gccttcttat tcggccttga attgatcata tgcggattag aaaaacaact
     3061 taaatgtgaa agtgggtctt aaggcatcaa ataaaacgaa aggctcagtc gaaagactgg
     3121 gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc tgagtaggac aaatccgccg
     3181 ccctagacct agggcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     3241 cggttatcca cagaatcagg ggataacgca tgcaagaaca tgtgagcaaa aggccagcaa
     3301 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     3361 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     3421 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     3481 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     3541 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     3601 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     3661 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     3721 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     3781 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     3841 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     3901 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     3961 gctcagtgga acgaaaactc acgttaaggg attttggtca tgactagtgc ttggattctc
     4021 accaataaaa aacgcccggc ggcaaccgag cgttctgaac aaatccagat ggagttctga
     4081 ggtcattact ggatctatca acaggagtcc aagcgagctc gatatcaaat tacgccccgc
     4141 cctgccactc atcgcagtac tgttgtaatt cattaagcat tctgccgaca tggaagccat
     4201 cacaaacggc atgatgaacc tgaatcgcca gcggcatcag caccttgtcg ccttgcgtat
     4261 aatatttgcc catggtgaaa acgggggcga agaagttgtc catattggcc acgtttaaat
     4321 caaaactggt gaaactcacc cagggattgg ctgagacgaa aaacatattc tcaataaacc
     4381 ctttagggaa ataggccagg ttttcaccat aacacgccac atcttgcgaa tatatgtgta
     4441 gaaactgccg gaaatcgtcg tggtattcac tccagagcga tgaaaacgtt tcagtttgct
     4501 catggaaaac ggtgtaacaa gggtgaacac tatcccatat caccagctca ccgtctttca
     4561 ttgccatacg aaattccgga tgagcattca tcaggcgggc aagaatgtga ataaaggccg
     4621 gataaaactt gtgcttattt ttctttacgg tctttaaaaa ggccgtaata tccagctgaa
     4681 cggtctggtt ataggtacat tgagcaactg actgaaatgc ctcaaaatgt tctttacgat
     4741 gccattggga tatatcaacg gtggtatatc cagtgatttt tttctccatt ttagcttcct
     4801 tagctcctga aaatctcgat aactcaaaaa atacgaccgg
//
