LOCUS       sequence-249054-        3234 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes VPR activation domain as a Type 3a part to be used in the 
            MTK system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3234)
  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 3234)
  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: MTK3a_015
FEATURES             Location/Qualifiers
     source          1..3234
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      56..644
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     545..564
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             759..908
                     /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             915..935
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1365..1736
                     /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
                     EDFSSIADMDFSALLSQISS"
     CDS             1755..2324
                     /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"
     terminator      complement(2344..2378)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(2449..3108)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3023..3042
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
       61 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
      121 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
      181 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
      241 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
      301 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
      361 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
      421 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
      481 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
      541 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
      601 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
      661 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
      721 cccctgattc tgtggataac cgtagtcggt ctcatatgga tgctttagac gattttgact
      781 tagatatgct tggttcagac gcgttagacg acttcgacct agacatgtta ggctcagatg
      841 cattggacga cttcgattta gatatgttgg gctccgatgc cctagatgac tttgatctag
      901 atatgctagg tagtcccaaa aagaagagga aagtgggatc ccagtatctg cccgacacag
      961 atgatagaca ccgaatcgaa gagaaacgca agcgaacgta tgaaaccttc aaatcgatca
     1021 tgaagaaatc gcccttctcg ggtccgaccg atcccaggcc cccaccgaga aggattgcgg
     1081 tcccgtcccg ctcgtcggcc agcgtgccga agcctgcgcc gcagccctac cccttcacgt
     1141 cgagcctgag cacaatcaat tatgacgagt tcccgacgat ggtgttcccc tcgggacaaa
     1201 tctcacaagc ctcggcgctc gcaccagcgc ctccccaagt ccttccgcaa gcgcctgccc
     1261 cagcgcctgc accggcaatg gtgtccgccc tcgcacaggc ccctgcgccc gtccccgtgc
     1321 tcgcgcctgg accgccccag gcggtcgctc caccggctcc gaagccgacg caggccggag
     1381 agggaacact ctccgaagca cttcttcaac tccagtttga tgacgaggat cttggagcac
     1441 tccttggaaa ctcgacagac cctgcggtgt ttaccgacct cgcgtcagta gataactccg
     1501 aatttcagca gcttttgaac cagggtatcc cggtcgcgcc acatacaacg gagcccatgt
     1561 tgatggaata ccccgaagca atcacgagac ttgtgacggg agcgcagcgg cctcccgatc
     1621 ccgcacccgc acctttgggg gcacctggcc tccctaacgg acttttgagc ggcgacgagg
     1681 atttctcctc catcgccgat atggatttct cagccttgct gtcacagatt tccagcggct
     1741 ctggcagcgg cagccgggat tccagggaag ggatgttttt gccgaagcct gaggccggct
     1801 ccgctattag tgacgtgttt gagggccgcg aggtgtgcca gccaaaacga atccggccat
     1861 ttcatcctcc aggaagtcca tgggccaacc gcccactccc cgccagcctc gcaccaacac
     1921 caaccggtcc agtacatgag ccagtcgggt cactgacccc ggcaccagtc cctcagccac
     1981 tggatccagc gcccgcagtg actcccgagg ccagtcacct gttggaggat cccgatgaag
     2041 aaacgagcca ggctgtcaaa gcccttcggg agatggccga tactgtgatt ccccagaagg
     2101 aagaggctgc aatctgtggc caaatggacc tttcccatcc gcccccaagg ggccatctgg
     2161 atgagctgac aaccacactt gagtccatga ccgaggatct gaacctggac tcacccctga
     2221 ccccggaatt gaacgagatt ctggatacct tcctgaacga cgagtgcctc ttgcatgcca
     2281 tgcatatcag cacaggactg tccatcttcg acacatctct gtttggttct tgagaccaga
     2341 ccaataaaaa acgcccggcg gcaaccgagc gttctgaaca aatccagatg gagttctgag
     2401 gtcattactg gatctatcaa caggagtcca agcgagctcg atatcaaatt acgccccgcc
     2461 ctgccactca tcgcagtact gttgtaattc attaagcatt ctgccgacat ggaagccatc
     2521 acaaacggca tgatgaacct gaatcgccag cggcatcagc accttgtcgc cttgcgtata
     2581 atatttgccc atggtgaaaa cgggggcgaa gaagttgtcc atattggcca cgtttaaatc
     2641 aaaactggtg aaactcaccc agggattggc tgaaacgaaa aacatattct caataaaccc
     2701 tttagggaaa taggccaggt tttcaccgta acacgccaca tcttgcgaat atatgtgtag
     2761 aaactgccgg aaatcgtcgt ggtattcact ccagagcgat gaaaacgttt cagtttgctc
     2821 atggaaaacg gtgtaacaag ggtgaacact atcccatatc accagctcac cgtctttcat
     2881 tgccatacga aattccggat gagcattcat caggcgggca agaatgtgaa taaaggccgg
     2941 ataaaacttg tgcttatttt tctttacggt ctttaaaaag gccgtaatat ccagctgaac
     3001 ggtctggtta taggtacatt gagcaactga ctgaaatgcc tcaaaatgtt ctttacgatg
     3061 ccattgggat atatcaacgg tggtatatcc agtgattttt ttctccattt tagcttcctt
     3121 agctcctgaa aatctcgata actcaaaaaa tacgcccggt agtgatctta tttcattatg
     3181 gtgaaagttg gaacctctta cgtgcccgat caatcatgac caaaatccct taac
//
