LOCUS       sequence-249733-        2409 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes Tet-On 3G transactivator protein as a Type 3b 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: MTK3b_029
FEATURES             Location/Qualifiers
     source          1..2409
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      77..665
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     566..585
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             780..1520
                     /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="SRLDKSKVINSALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHVK
                     NKRALLDALPIEMLDRHHTHSCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTRP
                     TEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTTD
                     SMPPLLKQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPAD
                     ALDDFDLDMLPADALDDFDLDMLPG"
     terminator      complement(1540..1574)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(1645..2304)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2219..2238
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
       61 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
      121 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
      181 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
      241 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
      301 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
      361 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
      421 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
      481 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
      541 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
      601 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
      661 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
      721 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtagtcgg tctcattctt
      781 ctagactgga caagagcaaa gtcataaact ctgctctgga attactcaat ggagtcggta
      841 tcgaaggcct gacgacaagg aaactcgctc aaaagctggg agttgagcag cctaccctgt
      901 actggcacgt gaagaacaag cgggccctgc tcgatgccct gccaatcgag atgctggaca
      961 ggcatcatac ccactcctgc cccctggaag gcgagtcatg gcaagacttt ctgcggaaca
     1021 acgccaagtc ataccgctgt gctctcctct cacatcgcga cggggctaaa gtgcatctcg
     1081 gcacccgccc aacagagaaa cagtacgaaa ccctggaaaa tcagctcgcg ttcctgtgtc
     1141 agcaaggctt ctccctggag aacgcactgt acgctctgtc cgccgtgggc cactttacac
     1201 tgggctgcgt attggaggaa caggagcatc aagtagcaaa agaggaaaga gagacaccta
     1261 ccaccgattc tatgccccca cttctgaaac aagcaattga gctgttcgac cggcagggag
     1321 ccgaacctgc cttccttttc ggcctggaac taatcatatg tggcctggag aaacagctaa
     1381 agtgcgaaag cggcgggccg accgacgccc ttgacgattt tgacttagac atgctcccag
     1441 ccgatgccct tgacgacttt gaccttgata tgctgcctgc tgacgctctt gacgattttg
     1501 accttgacat gctccccggg ggatcctgag accagaccaa taaaaaacgc ccggcggcaa
     1561 ccgagcgttc tgaacaaatc cagatggagt tctgaggtca ttactggatc tatcaacagg
     1621 agtccaagcg agctcgatat caaattacgc cccgccctgc cactcatcgc agtactgttg
     1681 taattcatta agcattctgc cgacatggaa gccatcacaa acggcatgat gaacctgaat
     1741 cgccagcggc atcagcacct tgtcgccttg cgtataatat ttgcccatgg tgaaaacggg
     1801 ggcgaagaag ttgtccatat tggccacgtt taaatcaaaa ctggtgaaac tcacccaggg
     1861 attggctgaa acgaaaaaca tattctcaat aaacccttta gggaaatagg ccaggttttc
     1921 accgtaacac gccacatctt gcgaatatat gtgtagaaac tgccggaaat cgtcgtggta
     1981 ttcactccag agcgatgaaa acgtttcagt ttgctcatgg aaaacggtgt aacaagggtg
     2041 aacactatcc catatcacca gctcaccgtc tttcattgcc atacgaaatt ccggatgagc
     2101 attcatcagg cgggcaagaa tgtgaataaa ggccggataa aacttgtgct tatttttctt
     2161 tacggtcttt aaaaaggccg taatatccag ctgaacggtc tggttatagg tacattgagc
     2221 aactgactga aatgcctcaa aatgttcttt acgatgccat tgggatatat caacggtggt
     2281 atatccagtg atttttttct ccattttagc ttccttagct cctgaaaatc tcgataactc
     2341 aaaaaatacg cccggtagtg atcttatttc attatggtga aagttggaac ctcttacgtg
     2401 cccgatcaa
//
