LOCUS       sequence-239121-        4757 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 4757)
  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 4757)
  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..4757
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      6..92
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(256..844)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(336..355)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(932..1026)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1047..1706)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1621..1640
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(1707..1809)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     1875..1893
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(1914..1932,1933..1938,1939..1955,1956..1961,
                     1962..1987)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    1914..1932
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1939..1957
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      2757..2828
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2844..2871
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             3138..3149
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             3156..3869
                     /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      3896..3967
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3983..4010
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        4062..4096
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     CDS             4131..4757
                     /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(4207..4226)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
ORIGIN
        1 ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt
       61 tgtcggtgaa cgctctcctg agtaggacaa atccgccgcc ctagacctag gcgttcggct
      121 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
      181 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
      241 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
      301 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
      361 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
      421 tctcccttcg ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc tcagttcggt
      481 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
      541 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
      601 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
      661 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
      721 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
      781 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
      841 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
      901 ttaagggatt ttggtcatga ctagtgcttg gattctcacc aataaaaaac gcccggcggc
      961 aaccgagcgt tctgaacaaa tccagatgga gttctgaggt cattactgga tctatcaaca
     1021 ggagtccaag cgagctcgat atcaaattac gccccgccct gccactcatc gcagtactgt
     1081 tgtaattcat taagcattct gccgacatgg aagccatcac agacggcatg atgaacctga
     1141 atcgccagcg gcatcagcac cttgtcgcct tgcgtataat atttgcccat ggtgaaaacg
     1201 ggggcgaaga agttgtccat attggccacg tttaaatcaa aactggtgaa actcacccag
     1261 ggattggctg agacgaaaaa catattctca ataaaccctt tagggaaata ggccaggttt
     1321 tcaccgtaac acgccacatc ttgcgaatat atgtgtagaa actgccggaa atcgtcgtgg
     1381 tattcactcc agagcgatga aaacgtttca gtttgctcat ggaaaacggt gtaacaaggg
     1441 tgaacactat cccatatcac cagctcaccg tctttcattg ccatacgaaa ttccggatga
     1501 gcattcatca ggcgggcaag aatgtgaata aaggccggat aaaacttgtg cttatttttc
     1561 tttacggtct ttaaaaaggc cgtaatatcc agctgaacgg tctggttata ggtacattga
     1621 gcaactgact gaaatgcctc aaaatgttct ttacgatgcc attgggatat atcaacggtg
     1681 gtatatccag tgattttttt ctccatttta gcttccttag ctcctgaaaa tctcgataac
     1741 tcaaaaaata cgcccggtag tgatcttatt tcattatggt gaaagttgga acctcttacg
     1801 tgccgatcaa cgtctcattt tcgccagata tcgacgtcta agaaaccatt attatcatga
     1861 cattaaccta taaaaatagg cgtatcacga ggccctttcg tcttcacctc gagtccctat
     1921 cagtgataga gattgacatc cctatcagtg atagagatac tgagcacatc agcaggacgc
     1981 actgaccgca taaaggactt agcaaagccc aattttaaac aaatgcaaga gaactacaag
     2041 attctggtgg tcgatgacga catgcgcctg cgtgcgctgc tggaacgtta tctcaccgaa
     2101 caaggcttcc aggttcgaag cgtcgctaat gcagaacaga tggatcgcct gctgactcgt
     2161 gaatctttcc atcttatggt actggattta atgttacctg gtgaagatgg cttgtcgatt
     2221 tgccgacgtc ttcgtagtca gagcaacccg atgccgatca ttatggtgac ggcgaaaggg
     2281 gaagaagtgg accgtatcgt aggcctggag attggcgctg acgactacat tccaaaaccg
     2341 tttaacccgc gtgaactgct ggcccgtatc cgtgcggtgc tgcgtcgtca aagccacgga
     2401 gaaaccatca cggaaaccct gggtgcagtc aaaaataatt tgctcagcgt caacaacgtt
     2461 agcctagatg tggccaatca ggtggcctat tgccagggtc aacgcattgc cctttcggaa
     2521 aaggaagtgg cgctgttaac cctattctta caagctcccg gacaaattct cagtcacgaa
     2581 gagatctatt cccacctttg gcccggcgaa agccccccca gtagtaacgt gctggcggcc
     2641 ctggtgcgat tactgcggcg caaaattgaa cagcccaatg ccccccgcct gattaattcc
     2701 gtttacggca aggggtactg ttttgaggcc aattagtaat aatctagacc aggcatcaaa
     2761 taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga
     2821 acgctctcta ctagagtcac actggctcac cttcgggtgg gcctttctgc gtttatagta
     2881 aatcactgca taattcgtgt cgctcaaggc gcactcccgt tctggataat gttttttgcg
     2941 ccgacatcat aacggttctg gcaaatattc tgaaatgagc tggttagcta gtcagtaaaa
     3001 agcagaaagt tttagtggtc agaaaaagcc cacgagctta ccagttggta aaagttaaga
     3061 aattttaact ccccgttttc actgcaatta tgagaagata atgaaaaatg agatgatcat
     3121 gaaagaccat tatcactaaa gaggagaaat actagatgcg taaaggcgaa gagctgttca
     3181 ctggtgtcgt ccctattctg gtggaactgg atggtgatgt caacggtcat aagttttccg
     3241 tgcgtggcga gggtgaaggt gacgcaacta atggtaaact gacgctgaag ttcatctgta
     3301 ctactggtaa actgccggta ccttggccga ctctggtaac gacgctgact tatggtgttc
     3361 agtgctttgc tcgttatccg gaccatatga agcagcatga cttcttcaag tccgccatgc
     3421 cggaaggcta tgtgcaggaa cgcacgattt cctttaagga tgacggcacg tacaaaacgc
     3481 gtgcggaagt gaaatttgaa ggcgataccc tggtaaaccg cattgagctg aaaggcattg
     3541 actttaaaga agacggcaat atcctgggcc ataagctgga atacaatttt aacagccaca
     3601 atgtttacat caccgccgat aaacaaaaaa atggcattaa agcgaatttt aaaattcgcc
     3661 acaacgtgga ggatggcagc gtgcagctgg ctgatcacta ccagcaaaac actccaatcg
     3721 gtgatggtcc tgttctgctg ccagacaatc actatctgag cacgcaaagc gttctgtcta
     3781 aagatccgaa cgagaaacgc gatcatatgg ttctgctgga gttcgtaacc gcagcgggca
     3841 tcacgcatgg tatggatgaa ctgtacaaat gatgataata atctagacca ggcatcaaat
     3901 aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa
     3961 cgctctctac tagagtcaca ctggctcacc ttcgggtggg cctttctgcg tttataggta
     4021 cccgtggatc ctctagagtc gacctgcagg catgcaagct ttttacggct agctcagtcc
     4081 taggtatagt gctagcccag ccagagaaac actctttaac agggggtagt atgatgtctc
     4141 gtttagataa aagtaaagtg attaacagcg cattagagct gcttaatgag gtcggaatcg
     4201 aaggtttaac aacccgtaaa ctcgcccaga agctaggtgt agagcagcct acattgtatt
     4261 ggcatgtaaa aaataagcgg gctttgctcg acgccttagc cattgagatg ttagataggc
     4321 accatactca cttttgccct ttagaagggg aaagctggca agatttttta cgtaataacg
     4381 ctaaaagttt tagatgtgct ttactaagtc atcgcgatgg agcaaaagta catttaggta
     4441 cacggcctac agaaaaacag tatgaaactc tcgaaaatca attagccttt ttatgccaac
     4501 aaggtttttc actagagaat gcattatatg cactcagcgc agtggggcat tttactttag
     4561 gttgcgtatt ggaagatcaa gagcatcaag tcgctaaaga agaaagggaa acacctacta
     4621 ctgatagtat gccgccatta ttacgacaag ctatcgaatt atttgatcac caaggtgcag
     4681 agccagcctt cttattcggc cttgaattga tcatatgcgg attagaaaaa caacttaaat
     4741 gtgaaagtgg gtcttaa
//
