LOCUS       sequence-437809-        6198 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tetracycline sensor v3 for Staphylococcus aureus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6198)
  AUTHORS   Rondthaler SN, Sarker B, Howitz N, Shah I, Andrews LB
  TITLE     Toolbox of Characterized Genetic Parts for Staphylococcus aureus.
  JOURNAL   ACS Synth Biol. 2023 Dec 8. doi: 10.1021/acssynbio.3c00325.
  PUBMED    38064657
REFERENCE   2  (bases 1 to 6198)
  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"; journalName: "ACS Synth 
            Biol. 2023 Dec 8. doi: 10.1021/acssynbio.3c00325."
COMMENT     Sequence Label: pSR1154
FEATURES             Location/Qualifiers
     source          1..6198
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1909..1926)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(1933..2646)
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Nager et al., 2011)"
                     /label=superfolder GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHVVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(1973..1993)
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     2588..2616
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     protein_bind    2789..2807
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     CDS             3198..3821
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /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="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     complement(3271..3290)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     CDS             complement(4578..4589)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acggagcatg
      601 ggaattcgag ctgatatttt tgactaaacc aaatgctaac ccagaaatac aatcactgtg
      661 tctaatgaat aatttgtttt ataaacactt ttttgtttac ttctcatttt taattagtta
      721 taattaacta aataatagag cattaaatat atttaataaa acttatttaa tgcaaaatta
      781 tgactaacat atctgagggt tataatgaac gagaaaaata taaaacacag tcaaaacttt
      841 attacttcaa aacataatat agataaaata atgacaaata taagattaaa tgaacatgat
      901 aatatctttg aaatcggctc aggaaaaggc cattttaccc ttgaattagt aaagaggtgt
      961 aatttcgtaa ctgccattga aatagaccat aaattatgca aaactacaga aaataaactt
     1021 gttgatcacg ataatttcca agttttaaac aaggatatat tgcagtttaa atttcctaaa
     1081 aaccaatcct ataaaatata tggtaatata ccttataaca taagtacgga tataatacgc
     1141 aaaattgttt ttgatagtat agctaatgag atttatttaa tcgtggaata cgggtttgct
     1201 aaaagattat taaatacaaa acgctcattg gcattacttt taatggcaga agttgatatt
     1261 tctatattaa gtatggttcc aagagaatat tttcatccta aacctaaagt gaatagctca
     1321 cttatcagat taagtagaaa aaaatcaaga atatcacaca aagataaaca aaagtataat
     1381 tatttcgtta tgaaatgggt taacaaagaa tacaagaaaa tatttacaaa aaatcaattt
     1441 aacaattcct taaaacatgc aggaattgac gatttaaaca atattagctt tgaacaattc
     1501 ttatctcttt tcaatagcta taaattattt aataagtaag ttaagggatg ccgtagaaaa
     1561 gggaaatagg cgcgcctatt ctaatgcata ataaatactg ataacatctt atattttgta
     1621 ttatattttg tattatcgtt gacatgtata attttgatat caaaaactga ttttccctct
     1681 attattttcg agatttattt tcttaattct ctttaacaaa ctagaaatat tgtatataca
     1741 aaaaattata aataatagat gaatagttta attataggtg ttcatcaatc gaaaaagcaa
     1801 cgtatcttat ttaaagtgcg ttgctttttt ctcatttata aggttaaata attctcatat
     1861 atcaagcaaa gtgacaggcg atgcgctaaa catgattacg aattcttagt ggtggtggtg
     1921 gtggtgggat cctttgtaga gctcatccat gccatgtgta atcccagcag cagttacaaa
     1981 ctcaagaagg accacgtggt cacgcttttc gttaggatct ttcgaaagga cagattgtgt
     2041 cgacaggtaa tggttgtctg gtaaaaggac agggccatcg ccaattggag tattttgttg
     2101 ataatggtct gctagttgaa cggaaccatc ttcaacgttg tggcgaattt tgaagttagc
     2161 tttgattcca ttcttttgtt tgtctgccgt gatgtataca ttgtgtgagt taaagttgta
     2221 ctcgagtttg tgtccaagaa tgtttccatc ttctttaaaa tcaataccct ttaactcgat
     2281 acgattaaca agggtatcac cttcaaactt gacttcagca cgcgtcttgt aggtcccgtc
     2341 atctttgaaa gatatagtgc gttcctgtac ataaccttca ggcatggcac tcttgaaaaa
     2401 gtcatgacgt ttcatgtgat ccggataacg ggaaaagcat tgaacaccat aggtcagagt
     2461 agtgacaagt gttggccacg gaacaggtag ttttccagta gtgcaaataa atttaagggt
     2521 gagttttccg tttgtagcat caccttcacc ctctccacgg acagaaaatt tgtgcccatt
     2581 aacatcacca tctaattcaa caagaattgg gacaactcca gtgaaaagtt cttctccttt
     2641 gctcataaat aatcatcctc ctaaggtacc cggggatccg atgcatcttg ctcgatacat
     2701 ttgcccgata atatattgat atctaatctt tatttattat agatatgtta gtcataattt
     2761 tgcattaaag cacgcttatt ttaattatac tctatcaatg atagagtgtc aactcccaaa
     2821 ttattcatta gacacagtga ttgtatttct gggttagcat ttggtttagt caaaaatact
     2881 ccgatccgaa tcatgaatta caagcaaaag tagcggtgat tgttaaaatt gatggtaaac
     2941 aatcaccgct atttttgctt gtgtatgtat aaaaaaggga tcaaaggtca tcccccatga
     3001 ttgatagtgg ggggatgact tttgatccta tgttcatgtt gcttatttaa tcgcctttga
     3061 tcactttaaa ataccttaaa acccctggaa tttctggctt tgccagacct atcattttgg
     3121 agaaaagggc ttaaatgttt gctttcgttg aattttagat ttaaaatgaa gggtgcggtg
     3181 gaaggtagat gatcgacatg tccagattag ataaaagtaa agtgattaac agcgcattag
     3241 agctgcttaa tgaggtcgga atcgaaggtt taacaacccg taaactcgcc cagaagctag
     3301 gtgtagagca gcctacattg tattggcatg taaaaaataa gcgggctttg ctcgacgcct
     3361 tagccattga gatgttagat aggcaccata ctcacttttg ccctttagaa ggggaaagct
     3421 ggcaagattt tttacgtaat aacgctaaaa gttttagatg tgctttacta agtcatcgcg
     3481 atggagcaaa agtacattta ggtacacggc ctacagaaaa acagtatgaa actctcgaaa
     3541 atcaattagc ctttttatgc caacaaggtt tttcactaga gaatgcatta tatgcactca
     3601 gcgctgtggg gcattttact ttaggttgcg tattggaaga tcaagagcat caagtcgcta
     3661 aagaagaaag ggaaacacct actactgata gtatgccgcc attattacga caagctatcg
     3721 aattatttga tcaccaaggt gcagagccag ccttcttatt cggccttgaa ttgatcatat
     3781 gcggattaga aaaacaactt aaatgtgaaa gtgggtccta attagtcggt accaaattcc
     3841 agaaaagaga cgctttcgag cgtctttttt cgttttggtc ccacgaattc tccttatgct
     3901 cttacggagt ttttagagaa aaattaaaaa ttctcgattt ttgataaaaa acgccctgca
     3961 ggaatttaga aaaacatgtg gaagttttaa aggattttat gctaattttt aatttgcatg
     4021 taactcgagg ggaatatttg aggggatttt gaaacgagtt tcttcttgtt ttcacactgt
     4081 ttttttattc ctattggtgt tgttgcttac tttttgtttt tctataaaga tgatgctttt
     4141 gacgagatgg aagaaaaata tgattatcat gaagataata aaaaatagac gacgcattta
     4201 tgccgagaaa atttattgat gttgagaaga acccttaact aaacttgcag acgaatgtcg
     4261 gcatagcgtg agctattaag ccgaccattc gacaagtttt gggattgtta agggttccga
     4321 ggctcaacgt caataaagca attggaataa atagtatcta aagtaacccg tatttaagat
     4381 tacattgtac atccttcgaa accatagtag ctgcgattcc agctaggttt ccttttatcc
     4441 caattttagc taattttttt gctacgccct ttaagttttt ggctttaaca tcttcaaaaa
     4501 ctgctgtgat tatgtttcca gttaaaagtt ctccatatga gtttaaaact tcagaataga
     4561 aacagtcacc gctatttttg tagttcccac caactgctct aggtgatatt gaactagttt
     4621 tctttttatt ctctatgtcg ttttttagtt tttggaatcc ttcaggaaca tatccatatc
     4681 gagatttggc cgattttatt tgataatcct tttgactttt tagaaagttt tatttattca
     4741 ttggttttca cattaattta tttaggttta gagggtcata aaaagaagaa ggaataggtt
     4801 gttttttgaa acgagtgtga acgagtttct tcttgtcttg atactatata gaaataactc
     4861 gattttatat atatagctgt aactgttgat attacagtgt ttaaacgtgt ttttgtgcgt
     4921 gaaaggaaaa tttgacaata aaaaacccca gttatattat taaggtgtcg aatcttaaat
     4981 aatactgggg gtctttttat atgcaatata atactactag aagtataacc gaaaatcaag
     5041 ataataaaac gttaaaagat atgacgaaaa gtgggaaaca acgcccatgg agagaaaaga
     5101 aaatagataa tgtaagctat gcagatatac tagaaatttt aaaaatcaaa aaggctttta
     5161 atgtaaaaca atgtggtaat attttagaat ttaagccaac tgatgaaggc tatttgaagt
     5221 tacataagac atggttttgt aaatcaaaat tatgtccggt ttgtaattgg agacgtgcta
     5281 tgaaaaatag ttatcaagct caaaaagtga ttgaaaaagt aattaaggaa aagccaaaag
     5341 cacgttggtt gtttttaaca ctttcaacaa aaaatgcgat agatggagat actttagaac
     5401 aaagtttgaa gcatctaact aaagcatttg ataggttgag tagatataaa aaggttaaac
     5461 aaaatcttgt tggatttatg cgttcaacag aagttaccgt taataaaaat gacggtagtt
     5521 ataatcagca catgcatgtt ttgttatgtg ttgaaaatgc atattttaga aaaaaagaga
     5581 attatataac tcaagaagaa tgggttaatt tatggcaaag agcattacaa gttgactatc
     5641 gacctgttgc taatgttaaa gcgatcaaac cgaatagaaa aggcgataaa gacattgaat
     5701 cggcaatcaa agagacctca aaatattcgg ttaaatcatc tgatttttta actgatgatg
     5761 atgaaaaaaa tcaagaaatt gtaagtgatt tagaaaaagg tttgtatcga aaacgtatgt
     5821 taagttatgg tggattgctt aaacaaaaac ataaaatttt aaacttagac gatgtcgaag
     5881 atggtaattt gattaatgca agtgatgaag ataaaacaac agacgaagaa gaaaaagcac
     5941 attcaattac cgcaatttgg aatttcgaaa agcaaaatta ttatttaaga cattagtgtt
     6001 gactaatgtc ttttttgttg attttttata aaaaagtact gtcttatttt tgtgacaaat
     6061 gctgtatgta gtgtcacaaa aataagacaa acgcaatata ttgtgtcaca aaaataagac
     6121 agtacagctt tgtatgatcc gtcgacgaaa gcctggccac gatgcgtccg gcgtagagcg
     6181 tgagatcaaa ggatcttc
//
