LOCUS       sequence-247812-        4737 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of ompR(1-137aa)-psdR(134-237aa) chimera under PLtetO-1, 
            output Promoter PpsdA110.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4737)
  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 4737)
  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: pSR240
FEATURES             Location/Qualifiers
     source          1..4737
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     73..91
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(112..130,131..136,137..153,154..159,160..185)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    112..130
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    137..155
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      967..1038
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1054..1081
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             1315..1326
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1333..2046
                     /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      2073..2144
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2160..2187
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        2239..2273
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     CDS             2308..2934
                     /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(2384..2403)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      2940..3026
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(3191..3779)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3271..3290)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3867..3961)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3982..4641)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4556..4575
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 ccggccaacg tctcattttc gccagatatc gacgtctaag aaaccattat tatcatgaca
       61 ttaacctata aaaataggcg tatcacgagg ccctttcgtc ttcacctcga gtccctatca
      121 gtgatagaga ttgacatccc tatcagtgat agagatactg agcacatcag caggacgcac
      181 tgaccgcata aaggacttag caaagcccaa ttttaaacaa atgcaagaga actacaagat
      241 tctggtggtc gatgacgaca tgcgcctgcg tgcgctgctg gaacgttatc tcaccgaaca
      301 aggcttccag gttcgaagcg tcgctaatgc agaacagatg gatcgcctgc tgactcgtga
      361 atctttccat cttatggtac tggatttaat gttacctggt gaagatggct tgtcgatttg
      421 ccgacgtctt cgtagtcaga gcaacccgat gccgatcatt atggtgacgg cgaaagggga
      481 agaagtggac cgtatcgtag gcctggagat tggcgctgac gactacattc caaaaccgtt
      541 taacccgcgt gaactgctgg cccgtatccg tgcggtgctg cgtcgtcagg cgaacgaact
      601 gccaggcgca ccgtcacagg aagaggcggt agtggagctt ggcgggctga cgatttatcc
      661 tgatcagaat gaggccgaat ggaacagcgt ccggattttg ttttcccaaa aagaattcca
      721 gcttctctcg atttttgtga gggagcataa aaaaattgtc tctcgtgatg aactattgga
      781 ggcgttatgg gatgacgttg attttgtcga tgacaatacg ttaacggtca atgtcaatcg
      841 gctgcggaga aaactggaga atgcaggact gacagattgc atttccacaa tcagaggaca
      901 aggttatcaa tttcaagtca atcggaagga tgaagcggaa tgttaataat aatctagacc
      961 aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt
     1021 ttgtcggtga acgctctcta ctagagtcac actggctcac cttcgggtgg gcctttctgc
     1081 gtttatagta aatcactgca taattcgtgt cgctcaaggc gcactcccgt tctggataat
     1141 gttttttgcg ccgacatcat aacggttctg gcaaatattc tgaaatgagc tggttagcta
     1201 gtcacaccct cgtgaatgtg acagcattgt aagattgggg agcggaattg caagaaagtt
     1261 cgatgggagg atgctgactt cctttttata ataaagaaaa aggaggagca gaacaaagag
     1321 gagaaatact agatgcgtaa aggcgaagag ctgttcactg gtgtcgtccc tattctggtg
     1381 gaactggatg gtgatgtcaa cggtcataag ttttccgtgc gtggcgaggg tgaaggtgac
     1441 gcaactaatg gtaaactgac gctgaagttc atctgtacta ctggtaaact gccggtacct
     1501 tggccgactc tggtaacgac gctgacttat ggtgttcagt gctttgctcg ttatccggac
     1561 catatgaagc agcatgactt cttcaagtcc gccatgccgg aaggctatgt gcaggaacgc
     1621 acgatttcct ttaaggatga cggcacgtac aaaacgcgtg cggaagtgaa atttgaaggc
     1681 gataccctgg taaaccgcat tgagctgaaa ggcattgact ttaaagaaga cggcaatatc
     1741 ctgggccata agctggaata caattttaac agccacaatg tttacatcac cgccgataaa
     1801 caaaaaaatg gcattaaagc gaattttaaa attcgccaca acgtggagga tggcagcgtg
     1861 cagctggctg atcactacca gcaaaacact ccaatcggtg atggtcctgt tctgctgcca
     1921 gacaatcact atctgagcac gcaaagcgtt ctgtctaaag atccgaacga gaaacgcgat
     1981 catatggttc tgctggagtt cgtaaccgca gcgggcatca cgcatggtat ggatgaactg
     2041 tacaaatgat gataataatc tagaccaggc atcaaataaa acgaaaggct cagtcgaaag
     2101 actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctctactag agtcacactg
     2161 gctcaccttc gggtgggcct ttctgcgttt ataggtaccc gtggatcctc tagagtcgac
     2221 ctgcaggcat gcaagctttt tacggctagc tcagtcctag gtatagtgct agcccagcca
     2281 gagaaacact ctttaacagg gggtagtatg atgtctcgtt tagataaaag taaagtgatt
     2341 aacagcgcat tagagctgct taatgaggtc ggaatcgaag gtttaacaac ccgtaaactc
     2401 gcccagaagc taggtgtaga gcagcctaca ttgtattggc atgtaaaaaa taagcgggct
     2461 ttgctcgacg ccttagccat tgagatgtta gataggcacc atactcactt ttgcccttta
     2521 gaaggggaaa gctggcaaga ttttttacgt aataacgcta aaagttttag atgtgcttta
     2581 ctaagtcatc gcgatggagc aaaagtacat ttaggtacac ggcctacaga aaaacagtat
     2641 gaaactctcg aaaatcaatt agccttttta tgccaacaag gtttttcact agagaatgca
     2701 ttatatgcac tcagcgcagt ggggcatttt actttaggtt gcgtattgga agatcaagag
     2761 catcaagtcg ctaaagaaga aagggaaaca cctactactg atagtatgcc gccattatta
     2821 cgacaagcta tcgaattatt tgatcaccaa ggtgcagagc cagccttctt attcggcctt
     2881 gaattgatca tatgcggatt agaaaaacaa cttaaatgtg aaagtgggtc ttaaggcatc
     2941 aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg
     3001 tgaacgctct cctgagtagg acaaatccgc cgccctagac ctagggcgtt cggctgcggc
     3061 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     3121 catgcaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     3181 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     3241 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     3301 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     3361 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     3421 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     3481 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     3541 cagcaactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     3601 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
     3661 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     3721 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     3781 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     3841 ggattttggt catgactagt gcttggattc tcaccaataa aaaacgcccg gcggcaaccg
     3901 agcgttctga acaaatccag atggagttct gaggtcatta ctggatctat caacaggagt
     3961 ccaagcgagc tcgatatcaa attacgcccc gccctgccac tcatcgcagt actgttgtaa
     4021 ttcattaagc attctgccga catggaagcc atcacaaacg gcatgatgaa cctgaatcgc
     4081 cagcggcatc agcaccttgt cgccttgcgt ataatatttg cccatggtga aaacgggggc
     4141 gaagaagttg tccatattgg ccacgtttaa atcaaaactg gtgaaactca cccagggatt
     4201 ggctgagacg aaaaacatat tctcaataaa ccctttaggg aaataggcca ggttttcacc
     4261 ataacacgcc acatcttgcg aatatatgtg tagaaactgc cggaaatcgt cgtggtattc
     4321 actccagagc gatgaaaacg tttcagtttg ctcatggaaa acggtgtaac aagggtgaac
     4381 actatcccat atcaccagct caccgtcttt cattgccata cgaaattccg gatgagcatt
     4441 catcaggcgg gcaagaatgt gaataaaggc cggataaaac ttgtgcttat ttttctttac
     4501 ggtctttaaa aaggccgtaa tatccagctg aacggtctgg ttataggtac attgagcaac
     4561 tgactgaaat gcctcaaaat gttctttacg atgccattgg gatatatcaa cggtggtata
     4621 tccagtgatt tttttctcca ttttagcttc cttagctcct gaaaatctcg ataactcaaa
     4681 aaatacgacc ggtagtgatc ttatttcatt atggtgaaag ttggaacctc ttacggg
//
