LOCUS       sequence-248658-        2409 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes Tet-On 3G transactivator protein as a Type 3 part to be used
            in the MTK system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2409)
  AUTHORS   Fonseca JP, Bonny AR, Kumar GR, Ng AH, Town J, Wu QC, Aslankoohi E, 
            Chen SY, Dods G, Harrigan P, Osimiri LC, Kistler AL, El-Samad H
  TITLE     A Toolkit for Rapid Modular Construction of Biological Circuits in 
            Mammalian Cells.
  JOURNAL   ACS Synth Biol. 2019 Nov 15;8(11):2593-2606. doi: 
            10.1021/acssynbio.9b00322. Epub 2019 Nov 5.
  PUBMED    31686495
REFERENCE   2  (bases 1 to 2409)
  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: 
            "10.1021/acssynbio.9b00322"; journalName: "ACS Synth Biol"; date: 
            "2019-11-15- 15"; volume: "8"; issue: "11"; pages: "2593-2606"
COMMENT     Sequence Label: MTK3_010
FEATURES             Location/Qualifiers
     source          1..2409
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      151..739
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     640..659
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             851..1594
                     /codon_start=1
                     /product="modified rtTA protein that binds tightly to 
                     promoters containing the tet operator in the presence of 
                     doxycycline"
                     /label=Tet-On(R) 3G
                     /translation="MSRLDKSKVINSALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHSCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLKQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     terminator      complement(1614..1648)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(1719..2378)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2293..2312
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 aactcaaaaa atacgcccgg tagtgatctt atttcattat ggtgaaagtt ggaacctctt
       61 acgtgcccga tcaatcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
      121 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
      181 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
      241 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
      301 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
      361 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
      421 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
      481 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
      541 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
      601 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
      661 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
      721 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
      781 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtag
      841 tcggtctcat atgtctagac tggacaagag caaagtcata aactctgctc tggaattact
      901 caatggagtc ggtatcgaag gcctgacgac aaggaaactc gctcaaaagc tgggagttga
      961 gcagcctacc ctgtactggc acgtgaagaa caagcgggcc ctgctcgatg ccctgccaat
     1021 cgagatgctg gacaggcatc atacccactc ctgccccctg gaaggcgagt catggcaaga
     1081 ctttctgcgg aacaacgcca agtcataccg ctgtgctctc ctctcacatc gcgacggggc
     1141 taaagtgcat ctcggcaccc gcccaacaga gaaacagtac gaaaccctgg aaaatcagct
     1201 cgcgttcctg tgtcagcaag gcttctccct ggagaacgca ctgtacgctc tgtccgccgt
     1261 gggccacttt acactgggct gcgtattgga ggaacaggag catcaagtag caaaagagga
     1321 aagagagaca cctaccaccg attctatgcc cccacttctg aaacaagcaa ttgagctgtt
     1381 cgaccggcag ggagccgaac ctgccttcct tttcggcctg gaactaatca tatgtggcct
     1441 ggagaaacag ctaaagtgcg aaagcggcgg gccgaccgac gcccttgacg attttgactt
     1501 agacatgctc ccagccgatg cccttgacga ctttgacctt gatatgctgc ctgctgacgc
     1561 tcttgacgat tttgaccttg acatgctccc cgggggatcc tgagaccaga ccaataaaaa
     1621 acgcccggcg gcaaccgagc gttctgaaca aatccagatg gagttctgag gtcattactg
     1681 gatctatcaa caggagtcca agcgagctcg atatcaaatt acgccccgcc ctgccactca
     1741 tcgcagtact gttgtaattc attaagcatt ctgccgacat ggaagccatc acaaacggca
     1801 tgatgaacct gaatcgccag cggcatcagc accttgtcgc cttgcgtata atatttgccc
     1861 atggtgaaaa cgggggcgaa gaagttgtcc atattggcca cgtttaaatc aaaactggtg
     1921 aaactcaccc agggattggc tgaaacgaaa aacatattct caataaaccc tttagggaaa
     1981 taggccaggt tttcaccgta acacgccaca tcttgcgaat atatgtgtag aaactgccgg
     2041 aaatcgtcgt ggtattcact ccagagcgat gaaaacgttt cagtttgctc atggaaaacg
     2101 gtgtaacaag ggtgaacact atcccatatc accagctcac cgtctttcat tgccatacga
     2161 aattccggat gagcattcat caggcgggca agaatgtgaa taaaggccgg ataaaacttg
     2221 tgcttatttt tctttacggt ctttaaaaag gccgtaatat ccagctgaac ggtctggtta
     2281 taggtacatt gagcaactga ctgaaatgcc tcaaaatgtt ctttacgatg ccattgggat
     2341 atatcaacgg tggtatatcc agtgattttt ttctccattt tagcttcctt agctcctgaa
     2401 aatctcgat
//
