LOCUS       sequence-248660-        2493 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes HA-tagged PYLcs fused to VP16 activator domain 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 2493)
  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 2493)
  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_012
FEATURES             Location/Qualifiers
     source          1..2493
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     306..325
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             523..543
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             544..777
                     /codon_start=1
                     /gene="UL48"
                     /product="transcriptional activation domain of herpes 
                     simplex virus protein VP16 (Triezenberg et al., 1988; 
                     Cousens et al., 1989)"
                     /label=VP16 AD
                     /translation="APPTDVSLGDELHLDGEDVAMAHADALDDFDLDMLGDGDSPGPGF
                     TPHDSAPYGALDMADFEFEQMFTDALGIDEYGG"
     CDS             1318..1344
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     terminator      complement(1364..1398)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(1469..2128)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2043..2062
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      join(2310..2493,1..405)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
       61 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
      121 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
      181 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
      241 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
      301 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
      361 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
      421 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
      481 tcccctgatt ctgtggataa ccgtagtcgg tctcatatgg gccctaaaaa gaagcgtaaa
      541 gtcgcccccc cgaccgatgt cagcctgggg gacgagctcc acttagacgg cgaggacgtg
      601 gcgatggcgc atgccgacgc gctagacgat ttcgatctgg acatgttggg ggacggggat
      661 tccccgggtc cgggatttac cccccacgac tccgccccct acggcgctct ggatatggcc
      721 gacttcgagt ttgagcagat gtttaccgat gcccttggaa ttgacgagta cggtgggggc
      781 gcgccaactc aagacgaatt cacccaactc tcccaatcaa tcgccgagtt ccacacgtac
      841 caactcggta acggccgttg ctcatctctc ctagctcagc gaatccacgc gccgccggaa
      901 acagtatggt ccgtggtcag acgtttcgat aggccacaga tttacaaaca cttcatcaaa
      961 agctgtaacg tgagtgaaga tttcgagatg cgagtgggat gcacgcgcga cgtgaacgtg
     1021 ataagtggat taccggcgaa tacgtcacga gagagattag atctgttgga cgatgatcgg
     1081 agagtgactg ggtttagtat aaccggtggt gaacataggc tgaggaatta taaatcggtt
     1141 acgacggttc atagatttga gaaagaagaa gaagaagaaa ggatctggac cgttgttttg
     1201 gaatcttatg ttgttgatgt accggaaggt aattcggagg aagatacgag attgtttgct
     1261 gatacggtta ttagattgaa tcttcagaaa cttgcttcga tcactgaagc tatgaactac
     1321 ccatacgatg ttccagatta cgctggatcc tgagaccaga ccaataaaaa acgcccggcg
     1381 gcaaccgagc gttctgaaca aatccagatg gagttctgag gtcattactg gatctatcaa
     1441 caggagtcca agcgagctcg atatcaaatt acgccccgcc ctgccactca tcgcagtact
     1501 gttgtaattc attaagcatt ctgccgacat ggaagccatc acaaacggca tgatgaacct
     1561 gaatcgccag cggcatcagc accttgtcgc cttgcgtata atatttgccc atggtgaaaa
     1621 cgggggcgaa gaagttgtcc atattggcca cgtttaaatc aaaactggtg aaactcaccc
     1681 agggattggc tgaaacgaaa aacatattct caataaaccc tttagggaaa taggccaggt
     1741 tttcaccgta acacgccaca tcttgcgaat atatgtgtag aaactgccgg aaatcgtcgt
     1801 ggtattcact ccagagcgat gaaaacgttt cagtttgctc atggaaaacg gtgtaacaag
     1861 ggtgaacact atcccatatc accagctcac cgtctttcat tgccatacga aattccggat
     1921 gagcattcat caggcgggca agaatgtgaa taaaggccgg ataaaacttg tgcttatttt
     1981 tctttacggt ctttaaaaag gccgtaatat ccagctgaac ggtctggtta taggtacatt
     2041 gagcaactga ctgaaatgcc tcaaaatgtt ctttacgatg ccattgggat atatcaacgg
     2101 tggtatatcc agtgattttt ttctccattt tagcttcctt agctcctgaa aatctcgata
     2161 actcaaaaaa tacgcccggt agtgatctta tttcattatg gtgaaagttg gaacctctta
     2221 cgtgcccgat caatcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     2281 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     2341 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     2401 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     2461 ctagtgtagc cgtagttagg ccaccacttc aag
//
