LOCUS       sequence-346736-        3244 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid encoding VPR (VP64 - SV40 NLS - p65 - Rta) as a Type 4a part
            to be used in the Dueber YTK system (MoClo kit).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3244)
  AUTHORS   Camara E, Lenitz I, Nygard Y
  TITLE     A CRISPR activation and interference toolkit for industrial 
            Saccharomyces cerevisiae strain KE6-12.
  JOURNAL   Sci Rep. 2020 Sep 3;10(1):14605. doi: 10.1038/s41598-020-71648-w.
  PUBMED    32884066
REFERENCE   2  (bases 1 to 3244)
  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: "Sci Rep."; 
            date: "2020-09-3"; volume: "10(1):14605. doi"; pages: " 
            10.1038/s41598-020-71648-w"
COMMENT     Sequence Label: EC0_4_VPR
FEATURES             Location/Qualifiers
     source          1..3244
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(11..45)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(116..775)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     690..709
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      957..1545
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1446..1465
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1660..1809
                     /codon_start=1
                     /product="tetrameric repeat of the minimal activation 
                     domain of herpes simplex virus VP16 (Beerli et al., 1998)"
                     /label=VP64
                     /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
                     DLDML"
     CDS             1834..1854
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2284..2640
                     /codon_start=1
                     /product="transcriptional activation domain of human RelA, 
                     also known as p65 (O'Shea and Perkins, 2008)"
                     /label=RelA (p65) AD
                     /translation="PTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNS
                     EFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGD
                     EDFSSIADMDFSALL"
     CDS             2659..3228
                     /codon_start=1
                     /product="transcriptional activation domain from the human 
                     herpesvirus 4 (Epstein-Barr virus) replication and 
                     transcription activator Rta/BRLF1 (Hardwick et al., 1992; 
                     Chavez et al., 2015)"
                     /label=Rta AD
                     /translation="RDSREGMFLPKPEAGSAISDVFEGREVCQPKRIRPFHPPGSPWAN
                     RPLPASLAPTPTGPVHEPVGSLTPAPVPQPLDPAPAVTPEASHLLEDPDEETSQAVKAL
                     REMADTVIPQKEEAAICGQMDLSHPPPRGHLDELTTTLESMTEDLNLDSPLTPELNEIL
                     DTFLNDECLLHAMHISTGLSIFDTSLF"
ORIGIN
        1 gaccagacca ataaaaaacg cccggcggca accgagcgtt ctgaacaaat ccagatggag
       61 ttctgaggtc attactggat ctatcaacag gagtccaagc gagctcgata tcaaattacg
      121 ccccgccctg ccactcatcg cagtactgtt gtaattcatt aagcattctg ccgacatgga
      181 agccatcaca aacggcatga tgaacctgaa tcgccagcgg catcagcacc ttgtcgcctt
      241 gcgtataata tttgcccatg gtgaaaacgg gggcgaagaa gttgtccata ttggccacgt
      301 ttaaatcaaa actggtgaaa ctcacccagg gattggctga aacgaaaaac atattctcaa
      361 taaacccttt agggaaatag gccaggtttt caccgtaaca cgccacatct tgcgaatata
      421 tgtgtagaaa ctgccggaaa tcgtcgtggt attcactcca gagcgatgaa aacgtttcag
      481 tttgctcatg gaaaacggtg taacaagggt gaacactatc ccatatcacc agctcaccgt
      541 ctttcattgc catacgaaat tccggatgag cattcatcag gcgggcaaga atgtgaataa
      601 aggccggata aaacttgtgc ttatttttct ttacggtctt taaaaaggcc gtaatatcca
      661 gctgaacggt ctggttatag gtacattgag caactgactg aaatgcctca aaatgttctt
      721 tacgatgcca ttgggatata tcaacggtgg tatatccagt gatttttttc tccattttag
      781 cttccttagc tcctgaaaat ctcgataact caaaaaatac gcccggtagt gatcttattt
      841 cattatggtg aaagttggaa cctcttacgt gcccgatcaa tcatgaccaa aatcccttaa
      901 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
      961 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     1021 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     1081 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     1141 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     1201 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     1261 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     1321 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     1381 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     1441 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     1501 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     1561 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     1621 tcccctgatt ctgtggataa ccgtagtcgg tctcaatccg acgcattgga cgattttgat
     1681 ctggatatgc tgggaagtga cgccctcgat gattttgacc ttgacatgct tggttcggat
     1741 gcccttgatg actttgacct cgacatgctc ggcagtgacg cccttgatga tttcgacctg
     1801 gacatgctga ttaactctag aagttccgga tctccgaaaa agaaacgcaa agttggtagc
     1861 cagtacctgc ccgacaccga cgaccggcac cggatcgagg aaaagcggaa gcggacctac
     1921 gagacattca agagcatcat gaagaagtcc cccttcagcg gccccaccga ccctagacct
     1981 ccacctagaa gaatcgccgt gcccagcaga tccagcgcca gcgtgccaaa acctgccccc
     2041 cagccttacc ccttcaccag cagcctgagc accatcaact acgacgagtt ccctaccatg
     2101 gtgttcccca gcggccagat ctctcaggcc tctgctctgg ctccagcccc tcctcaggtg
     2161 ctgcctcagg ctcctgctcc tgcaccagct ccagccatgg tgtctgcact ggctcaggca
     2221 ccagcacccg tgcctgtgct ggctcctgga cctccacagg ctgtggctcc accagcccct
     2281 aaacctacac aggccggcga gggcacactg tctgaagctc tgctgcagct gcagttcgac
     2341 gacgaggatc tgggagccct gctgggaaac agcaccgatc ctgccgtgtt caccgacctg
     2401 gccagcgtgg acaacagcga gttccagcag ctgctgaacc agggcatccc tgtggcccct
     2461 cacaccaccg agcccatgct gatggaatac cccgaggcca tcacccggct cgtgacaggc
     2521 gctcagaggc ctcctgatcc agctcctgcc cctctgggag caccaggcct gcctaatgga
     2581 ctgctgtctg gcgacgagga cttcagctct atcgccgaca tggacttctc cgcactgctg
     2641 ggtagcggat cgggatctcg ggattccagg gaagggatgt ttttgccgaa gcctgaggcc
     2701 ggctccgcta ttagtgacgt gtttgagggc cgcgaggtgt gccagccaaa acgaatccgg
     2761 ccatttcatc ctccaggaag tccatgggcc aaccgcccac tccccgccag cctcgcacca
     2821 acaccaaccg gtccagtaca tgagccagtc gggtcactga ccccggcacc agtccctcag
     2881 ccactggatc cagcgcccgc agtgactccc gaggccagtc acctgttgga ggatcccgat
     2941 gaagaaacga gccaggctgt caaagccctt cgggagatgg ccgatactgt gattccccag
     3001 aaggaagagg ctgcaatctg tggccaaatg gacctttccc atccgccccc aaggggccat
     3061 ctggatgagc tgacaaccac acttgagtcc atgaccgagg atctgaacct ggactcaccc
     3121 ctgaccccgg aattgaacga gattctggat accttcctga acgacgagtg cctcttgcat
     3181 gccatgcata tcagcacagg actgtccatc ttcgacacat ctctgtttta actcgagtgg
     3241 ctga
//
