LOCUS       sequence-247832-        4758 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of ompR(1-122aa)-manR(120-234aa) chimera under PLtetO-1, 
            output Promoter PmntCAB143.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4758)
  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 4758)
  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: pSR284
FEATURES             Location/Qualifiers
     source          1..4758
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     112..130
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(151..169,170..175,176..192,193..198,199..224)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    151..169
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    176..194
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      994..1065
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1081..1108
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             1375..1386
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1393..2106
                     /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      2133..2204
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2220..2247
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        2299..2333
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     CDS             2368..2994
                     /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(2444..2463)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      3000..3086
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(3251..3839)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3331..3350)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3927..4020)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(4042..4701)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4616..4635
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 tcttatttca ttatggtgaa agttggaacc tcttacgggc cggccaacgt ctcattttcg
       61 ccagatatcg acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt
      121 atcacgaggc cctttcgtct tcacctcgag tccctatcag tgatagagat tgacatccct
      181 atcagtgata gagatactga gcacatcagc aggacgcact gaccgcataa aggacttagc
      241 aaagcccaat tttaaacaaa tgcaagagaa ctacaagatt ctggtggtcg atgacgacat
      301 gcgcctgcgt gcgctgctgg aacgttatct caccgaacaa ggcttccagg ttcgaagcgt
      361 cgctaatgca gaacagatgg atcgcctgct gactcgtgaa tctttccatc ttatggtact
      421 ggatttaatg ttacctggtg aagatggctt gtcgatttgc cgacgtcttc gtagtcagag
      481 caacccgatg ccgatcatta tggtgacggc gaaaggggaa gaagtggacc gtatcgtagg
      541 cctggagatt ggcgctgacg actacattcc aaaaccgttt aacccgcgtg aactgctggc
      601 ccgtatccgt gcggtgctgc gtcgtcaaag ccacggagaa accatcacgg aaaccctggg
      661 tgcagtcaaa aataatttgc tcagcgtcaa caacgttagc ctagatgtgg ccaatcaggt
      721 ggcctattgc cagggtcaac gcattgccct ttcggaaaag gaagtggcgc tgttaaccct
      781 attcttacaa gctcccggac aaattctcag tcacgaagag atctattccc acctttggcc
      841 cggcgaaagc ccccccagta gtaacgtgct ggcggccctg gtgcgattac tgcggcgcaa
      901 aattgaacag cccaatgccc cccgcctgat taattccgtt tacggcaagg ggtactgttt
      961 tgaggccaat tagtaataat ctagaccagg catcaaataa aacgaaaggc tcagtcgaaa
     1021 gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctctacta gagtcacact
     1081 ggctcacctt cgggtgggcc tttctgcgtt tatagtaaat cactgcataa ttcgtgtcgc
     1141 tcaaggcgca ctcccgttct ggataatgtt ttttgcgccg acatcataac ggttctggca
     1201 aatattctga aatgagctgg ttagctagtc agtaaaaagc agaaagtttt agtggtcaga
     1261 aaaagcccac gagcttacca gttggtaaaa gttaagaaat tttaactccc cgttttcact
     1321 gcaattatga gaagataatg aaaaatgaga tgatcatgaa agaccattat cactaaagag
     1381 gagaaatact agatgcgtaa aggcgaagag ctgttcactg gtgtcgtccc tattctggtg
     1441 gaactggatg gtgatgtcaa cggtcataag ttttccgtgc gtggcgaggg tgaaggtgac
     1501 gcaactaatg gtaaactgac gctgaagttc atctgtacta ctggtaaact gccggtacct
     1561 tggccgactc tggtaacgac gctgacttat ggtgttcagt gctttgctcg ttatccggac
     1621 catatgaagc agcatgactt cttcaagtcc gccatgccgg aaggctatgt gcaggaacgc
     1681 acgatttcct ttaaggatga cggcacgtac aaaacgcgtg cggaagtgaa atttgaaggc
     1741 gataccctgg taaaccgcat tgagctgaaa ggcattgact ttaaagaaga cggcaatatc
     1801 ctgggccata agctggaata caattttaac agccacaatg tttacatcac cgccgataaa
     1861 caaaaaaatg gcattaaagc gaattttaaa attcgccaca acgtggagga tggcagcgtg
     1921 cagctggctg atcactacca gcaaaacact ccaatcggtg atggtcctgt tctgctgcca
     1981 gacaatcact atctgagcac gcaaagcgtt ctgtctaaag atccgaacga gaaacgcgat
     2041 catatggttc tgctggagtt cgtaaccgca gcgggcatca cgcatggtat ggatgaactg
     2101 tacaaatgat gataataatc tagaccaggc atcaaataaa acgaaaggct cagtcgaaag
     2161 actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctctactag agtcacactg
     2221 gctcaccttc gggtgggcct ttctgcgttt ataggtaccc gtggatcctc tagagtcgac
     2281 ctgcaggcat gcaagctttt tacggctagc tcagtcctag gtatagtgct agcccagcca
     2341 gagaaacact ctttaacagg gggtagtatg atgtctcgtt tagataaaag taaagtgatt
     2401 aacagcgcat tagagctgct taatgaggtc ggaatcgaag gtttaacaac ccgtaaactc
     2461 gcccagaagc taggtgtaga gcagcctaca ttgtattggc atgtaaaaaa taagcgggct
     2521 ttgctcgacg ccttagccat tgagatgtta gataggcacc atactcactt ttgcccttta
     2581 gaaggggaaa gctggcaaga ttttttacgt aataacgcta aaagttttag atgtgcttta
     2641 ctaagtcatc gcgatggagc aaaagtacat ttaggtacac ggcctacaga aaaacagtat
     2701 gaaactctcg aaaatcaatt agccttttta tgccaacaag gtttttcact agagaatgca
     2761 ttatatgcac tcagcgcagt ggggcatttt actttaggtt gcgtattgga agatcaagag
     2821 catcaagtcg ctaaagaaga aagggaaaca cctactactg atagtatgcc gccattatta
     2881 cgacaagcta tcgaattatt tgatcaccaa ggtgcagagc cagccttctt attcggcctt
     2941 gaattgatca tatgcggatt agaaaaacaa cttaaatgtg aaagtgggtc ttaaggcatc
     3001 aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg
     3061 tgaacgctct cctgagtagg acaaatccgc cgccctagac ctagggcgtt cggctgcggc
     3121 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     3181 catgcaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     3241 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     3301 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     3361 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     3421 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     3481 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     3541 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     3601 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     3661 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
     3721 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     3781 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     3841 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     3901 ggattttggt catgactagt gcttggattc tcaccaataa aaaacgcccg gcggcaaccg
     3961 agcgttctga acaaatccag atggagttct gaggtcatta ctggatctat caacaggagt
     4021 acaagcgagc tcgatatcaa attacgcccc gccctgccac tcatcgcagt actgttgtaa
     4081 ttcattaagc attctgccga catggaagcc atcacaaacg gcatgatgaa cctgaatcgc
     4141 cagcggcatc agcaccttgt cgccttgcgt ataatatttg cccatggtga aaacgggggc
     4201 gaagaagttg tccatattgg ccacgtttaa atcaaaactg gtgaaactca cccagggatt
     4261 ggctgagacg aaaaacatat tctcaataaa ccctttaggg aaataggcca ggttttcacc
     4321 ataacacgcc acatcttgcg aatatatgtg tagaaactgc cggaaatcgt cgtggtattc
     4381 actccagagc gatgaaaacg tttcagtttg ctcatggaaa acggtgtaac aagggtgaac
     4441 actatcccat atcaccagct caccgtcttt cattgccata cgaaattccg gatgagcatt
     4501 catcaggcgg gcaagaatgt gaataaaggc cggataaaac ttgtgcttat ttttctttac
     4561 ggtctttaaa aaggccgtaa tatccagctg aacggtctgg ttataggtac attgagcaac
     4621 tgactgaaat gcctcaaaat gttctttacg atgccattgg gatatatcaa cggtggtata
     4681 tccagtgatt tttttctcca ttttagcttc cttagctcct gaaaatctcg ataactcaaa
     4741 aaatacggcc ggtagtga
//
