LOCUS       sequence-240695-        2322 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TetR bacterial repressor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2322)
  AUTHORS   Shaw WM, Yamauchi H, Mead J, Gowers GF, Bell DJ, Oling D, Larsson N,
            Wigglesworth M, Ladds G, Ellis T
  TITLE     Engineering a Model Cell for Rational Tuning of GPCR Signaling.
  JOURNAL   Cell. 2019 Apr 18;177(3):782-796.e27. doi: 
            10.1016/j.cell.2019.02.023. Epub 2019 Apr 4.
  PUBMED    30955892
REFERENCE   2  (bases 1 to 2322)
  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: 
            "10.1016/j.cell.2019.02.023"; journalName: "Cell"; date: 
            "2019-04-18- 18"; volume: "177"; issue: "3"; pages: "782-796.e27"
COMMENT     Sequence Label: pWS931
FEATURES             Location/Qualifiers
     source          1..2322
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             13..633
                     /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(86..105)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     CDS             643..663
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      complement(689..723)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(794..1453)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1368..1387
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      1635..2223
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2124..2143
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agtcggtctc atatgtctag attagataaa agtaaagtga ttaacagcgc attagagctg
       61 cttaatgagg tcggaatcga aggtttaaca acccgtaaac tcgcccagaa gctaggtgta
      121 gagcagccta cattgtattg gcatgtaaaa aataagcggg ctttgctcga cgccttagcc
      181 attgagatgt tagataggca ccatactcac ttttgccctt tagaagggga aagctggcaa
      241 gattttttac gtaataacgc taaaagtttt agatgtgctt tactaagtca tcgcgatgga
      301 gcaaaagtac atttaggtac acggcctaca gaaaaacagt atgaaactct cgaaaatcaa
      361 ttagcctttt tatgccaaca aggtttttca ctagagaatg cattatatgc actcagcgct
      421 gtggggcatt ttactttagg ttgcgtattg gaagatcaag agcatcaagt cgctaaagaa
      481 gaaagggaaa cacctactac tgatagtatg ccgccattat tacgacaagc tatcgaatta
      541 tttgatcacc aaggtgcaga gccagccttc ttattcggcc ttgaattgat catatgcgga
      601 ttagaaaaac aacttaaatg tgaaagtgga tcaggttctc ctccaaaaaa aaagagaaaa
      661 gttggatctg gttcttgaga ccagaccaat aaaaaacgcc cggcggcaac cgagcgttct
      721 gaacaaatcc agatggagtt ctgaggtcat tactggatct atcaacagga gtccaagcga
      781 gctcgatatc aaattacgcc ccgccctgcc actcatcgca gtactgttgt aattcattaa
      841 gcattctgcc gacatggaag ccatcacaaa cggcatgatg aacctgaatc gccagcggca
      901 tcagcacctt gtcgccttgc gtataatatt tgcccatggt gaaaacgggg gcgaagaagt
      961 tgtccatatt ggccacgttt aaatcaaaac tggtgaaact cacccaggga ttggctgaaa
     1021 cgaaaaacat attctcaata aaccctttag ggaaataggc caggttttca ccgtaacacg
     1081 ccacatcttg cgaatatatg tgtagaaact gccggaaatc gtcgtggtat tcactccaga
     1141 gcgatgaaaa cgtttcagtt tgctcatgga aaacggtgta acaagggtga acactatccc
     1201 atatcaccag ctcaccgtct ttcattgcca tacgaaattc cggatgagca ttcatcaggc
     1261 gggcaagaat gtgaataaag gccggataaa acttgtgctt atttttcttt acggtcttta
     1321 aaaaggccgt aatatccagc tgaacggtct ggttataggt acattgagca actgactgaa
     1381 atgcctcaaa atgttcttta cgatgccatt gggatatatc aacggtggta tatccagtga
     1441 tttttttctc cattttagct tccttagctc ctgaaaatct cgataactca aaaaatacgc
     1501 ccggtagtga tcttatttca ttatggtgaa agttggaacc tcttacgtgc ccgatcaatc
     1561 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     1621 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     1681 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     1741 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     1801 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     1861 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     1921 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     1981 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     2041 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     2101 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     2161 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     2221 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     2281 atgttctttc ctgcgttatc ccctgattct gtggataacc gt
//
