LOCUS       sequence-239119-        4789 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of ompR(1-137aa)-yxdJ(132-229aa) chimera under PLtetO-1, 
            output Promoter PyxdL181.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4789)
  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 4789)
  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: pSR241
FEATURES             Location/Qualifiers
     source          1..4789
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(151..739)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(231..250)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(827..921)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(942..1601)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1516..1535
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(1602..1704)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     1770..1788
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(1809..1827,1828..1833,1834..1850,1851..1856,
                     1857..1882)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    1809..1827
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1834..1852
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      2646..2717
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2733..2760
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             3065..3076
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             3083..3796
                     /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      3823..3894
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3910..3937
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        3989..4023
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     CDS             4058..4684
                     /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(4134..4153)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      4690..4776
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 cctaggcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
       61 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
      121 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
      181 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
      241 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
      301 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcaatgct cacgctgtag
      361 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
      421 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
      481 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
      541 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
      601 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
      661 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
      721 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
      781 gaacgaaaac tcacgttaag ggattttggt catgactagt gcttggattc tcaccaataa
      841 aaaacgcccg gcggcaaccg agcgttctga acaaatccag atggagttct gaggtcatta
      901 ctggatctat caacaggagt ccaagcgagc tcgatatcaa attacgcccc gccctgccac
      961 tcatcgcagt actgttgtaa ttcattaagc attctgccga catggaagcc atcacagacg
     1021 gcatgatgaa cctgaatcgc cagcggcatc agcaccttgt cgccttgcgt ataatatttg
     1081 cccatggtga aaacgggggc gaagaagttg tccatattgg ccacgtttaa atcaaaactg
     1141 gtgaaactca cccagggatt ggctgagacg aaaaacatat tctcaataaa ccctttaggg
     1201 aaataggcca ggttttcacc gtaacacgcc acatcttgcg aatatatgtg tagaaactgc
     1261 cggaaatcgt cgtggtattc actccagagc gatgaaaacg tttcagtttg ctcatggaaa
     1321 acggtgtaac aagggtgaac actatcccat atcaccagct caccgtcttt cattgccata
     1381 cgaaattccg gatgagcatt catcaggcgg gcaagaatgt gaataaaggc cggataaaac
     1441 ttgtgcttat ttttctttac ggtctttaaa aaggccgtaa tatccagctg aacggtctgg
     1501 ttataggtac attgagcaac tgactgaaat gcctcaaaat gttctttacg atgccattgg
     1561 gatatatcaa cggtggtata tccagtgatt tttttctcca ttttagcttc cttagctcct
     1621 gaaaatctcg ataactcaaa aaatacgccc ggtagtgatc ttatttcatt atggtgaaag
     1681 ttggaacctc ttacgtgccg atcaacgtct cattttcgcc agatatcgac gtctaagaaa
     1741 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtcttc
     1801 acctcgagtc cctatcagtg atagagattg acatccctat cagtgataga gatactgagc
     1861 acatcagcag gacgcactga ccgcataaag gacttagcaa agcccaattt taaacaaatg
     1921 caagagaact acaagattct ggtggtcgat gacgacatgc gcctgcgtgc gctgctggaa
     1981 cgttatctca ccgaacaagg cttccaggtt cgaagcgtcg ctaatgcaga acagatggat
     2041 cgcctgctga ctcgtgaatc tttccatctt atggtactgg atttaatgtt acctggtgaa
     2101 gatggcttgt cgatttgccg acgtcttcgt agtcagagca acccgatgcc gatcattatg
     2161 gtgacggcga aaggggaaga agtggaccgt atcgtaggcc tggagattgg cgctgacgac
     2221 tacattccaa aaccgtttaa cccgcgtgaa ctgctggccc gtatccgtgc ggtgctgcgt
     2281 cgtcaggcga acgaactgcc aggcgcaccg tcacaggaag aggcggtagt tgaatatgcc
     2341 ggcgttcagc tctttgtgga acggtttgaa ctgcgttttc aggatgaaaa aagtgagctt
     2401 tctaaaaaag aaagcaagct tttggaagtg ctgcttgagc ggggagaaaa ggtgacgagt
     2461 cgggaccgtc tcatggaaaa gacgtgggac accgacatat tcatcgatga taatacactt
     2521 aacgtgtata tcacgcggct cagaaaaaaa ctgcgggagc tgaatgcgcc tgtttctatt
     2581 gaagcggtgc ggggcgaagg ctaccagctg agggcgcagt catgataata atctagacca
     2641 ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt
     2701 tgtcggtgaa cgctctctac tagagtcaca ctggctcacc ttcgggtggg cctttctgcg
     2761 tttatagtaa atcactgcat aattcgtgtc gctcaaggcg cactcccgtt ctggataatg
     2821 ttttttgcgc cgacatcata acggttctgg caaatattct gaaatgagct ggttagctag
     2881 tcaggacatc agttcgattt tcatttctta caaaaatgta aggctgcgtt aagaaaacaa
     2941 catagcccgc tttgtttccc cttgataaca tggatttatg tcaaggggat ttttatgttg
     3001 aacgtcccgg cactacatga aagcagccca gatcaacaag aataaaacca taaggaggat
     3061 tttcaaagag gagaaatact agatgcgtaa aggcgaagag ctgttcactg gtgtcgtccc
     3121 tattctggtg gaactggatg gtgatgtcaa cggtcataag ttttccgtgc gtggcgaggg
     3181 tgaaggtgac gcaactaatg gtaaactgac gctgaagttc atctgtacta ctggtaaact
     3241 gccggtacct tggccgactc tggtaacgac gctgacttat ggtgttcagt gctttgctcg
     3301 ttatccggac catatgaagc agcatgactt cttcaagtcc gccatgccgg aaggctatgt
     3361 gcaggaacgc acgatttcct ttaaggatga cggcacgtac aaaacgcgtg cggaagtgaa
     3421 atttgaaggc gataccctgg taaaccgcat tgagctgaaa ggcattgact ttaaagaaga
     3481 cggcaatatc ctgggccata agctggaata caattttaac agccacaatg tttacatcac
     3541 cgccgataaa caaaaaaatg gcattaaagc gaattttaaa attcgccaca acgtggagga
     3601 tggcagcgtg cagctggctg atcactacca gcaaaacact ccaatcggtg atggtcctgt
     3661 tctgctgcca gacaatcact atctgagcac gcaaagcgtt ctgtctaaag atccgaacga
     3721 gaaacgcgat catatggttc tgctggagtt cgtaaccgca gcgggcatca cgcatggtat
     3781 ggatgaactg tacaaatgat gataataatc tagaccaggc atcaaataaa acgaaaggct
     3841 cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctctactag
     3901 agtcacactg gctcaccttc gggtgggcct ttctgcgttt ataggtaccc gtggatcctc
     3961 tagagtcgac ctgcaggcat gcaagctttt tacggctagc tcagtcctag gtatagtgct
     4021 agcccagcca gagaaacact ctttaacagg gggtagtatg atgtctcgtt tagataaaag
     4081 taaagtgatt aacagcgcat tagagctgct taatgaggtc ggaatcgaag gtttaacaac
     4141 ccgtaaactc gcccagaagc taggtgtaga gcagcctaca ttgtattggc atgtaaaaaa
     4201 taagcgggct ttgctcgacg ccttagccat tgagatgtta gataggcacc atactcactt
     4261 ttgcccttta gaaggggaaa gctggcaaga ttttttacgt aataacgcta aaagttttag
     4321 atgtgcttta ctaagtcatc gcgatggagc aaaagtacat ttaggtacac ggcctacaga
     4381 aaaacagtat gaaactctcg aaaatcaatt agccttttta tgccaacaag gtttttcact
     4441 agagaatgca ttatatgcac tcagcgcagt ggggcatttt actttaggtt gcgtattgga
     4501 agatcaagag catcaagtcg ctaaagaaga aagggaaaca cctactactg atagtatgcc
     4561 gccattatta cgacaagcta tcgaattatt tgatcaccaa ggtgcagagc cagccttctt
     4621 attcggcctt gaattgatca tatgcggatt agaaaaacaa cttaaatgtg aaagtgggtc
     4681 ttaaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt
     4741 tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc cgccctaga
//
