LOCUS       sequence-167099-        5532 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway Donor vector containing DNMT3A , part of the Target 
            Accelerator Plasmid Collection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5532)
  AUTHORS   Kim E, Ilic N, Shrestha Y, Zou L, Kamburov A, Zhu C, Yang X, Lubonja
            R, Tran N, Nguyen C, Lawrence MS, Piccioni F, Bagul M, Doench JG, 
            Chouinard CR, Wu X, Hogstrom L, Natoli T, Tamayo P, Horn H, Corsello
            SM, Lage K, Root DE, Subramanian A, Golub TR, Getz G, Boehm JS, Hahn
            WC
  TITLE     Systematic Functional Interrogation of Rare Cancer Variants 
            Identifies Oncogenic Alleles.
  JOURNAL   Cancer Discov. 2016 Jul;6(7):714-26. doi: 
            10.1158/2159-8290.CD-16-0160. Epub 2016 May 4.
  PUBMED    27147599
REFERENCE   2  (bases 1 to 5532)
  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: 
            "10.1158/2159-8290.CD-16-0160"; journalName: "Cancer Discov"; date: 
            "2016-07"; volume: "6"; issue: "7"; pages: "714-26"
COMMENT     Sequence Label: pDONR223_DNMT3A_p.A910V
FEATURES             Location/Qualifiers
     source          1..5532
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     43..65
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     57..74
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     58..74
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    90..189
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             192..2930
                     /codon_start=1
                     /product="DNA methyltransferase 3A"
                     /label=DNMT3A
                     /translation="MPAMPSSGPGDTSSSAAEREEDRKDGEEQEEPRGKEERQEPSTTA
                     RKVGRPGRKRKHPPVESGDTPKDPAVISKSPSMAQDSGASELLPNGDLEKRSEPQPEEG
                     SPAGGQKGGAPAEGEGAAETLPEASRAVENGCCTPKEGRGAPAEAGKEQKETNIESMKM
                     EGSRGRLRGGLGWESSLRQRPMPRLTFQAGDPYYISKRKRDEWLARWKREAEKKAKVIA
                     GMNAVEENQGPGESQKVEEASPPAVQQPTDPASPTVATTPEPVGSDAGDKNATKAGDDE
                     PEYEDGRGFGIGELVWGKLRGFSWWPGRIVSWWMTGRSRAAEGTRWVMWFGDGKFSVVC
                     VEKLMPLSSFCSAFHQATYNKQPMYRKAIYEVLQVASSRAGKLFPVCHDSDESDTAKAV
                     EVQNKPMIEWALGGFQPSGPKGLEPPEEEKNPYKEVYTDMWVEPEAAAYAPPPPAKKPR
                     KSTAEKPKVKEIIDERTRERLVYEVRQKCRNIEDICISCGSLNVTLEHPLFVGGMCQNC
                     KNCFLECAYQYDDDGYQSYCTICCGGREVLMCGNNNCCRCFCVECVDLLVGPGAAQAAI
                     KEDPWNCYMCGHKGTYGLLRRREDWPSRLQMFFANNHDQEFDPPKVYPPVPAEKRKPIR
                     VLSLFDGIATGLLVLKDLGIQVDRYIASEVCEDSITVGMVRHQGKIMYVGDVRSVTQKH
                     IQEWGPFDLVIGGSPCNDLSIVNPARKGLYEGTGRLFFEFYRLLHDARPKEGDDRPFFW
                     LFENVVAMGVSDKRDISRFLESNPVMIDAKEVSAAHRARYFWGNLPGMNRPLASTVNDK
                     LELQECLEHGRIAKFSKVRTITTRSNSIKQGKDQHFPVFMNEKEDILWCTEMERVFGFP
                     VHYTDVSNMSRLARQRLLGRSWSVPVIRHLFAPLKEYFVCV"
     protein_bind    complement(2932..3031)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3048..3067)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3049..3067)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3072..3088)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3072..3088)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3519..4310
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      4403..4991
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4892..4911
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5145..5162
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(5321..5348)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(5440..5526)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ggcagttccc tactctcgcg ttaacgctag catggatgtt ttcccagtca cgacgttgta
       61 aaacgacggc cagtcttaag ctcgggcccc aaataatgat tttattttga ctgatagtga
      121 cctgttcgtt gcaacaaatt gatgagcaat gcttttttat aatgccaact ttgtacaaaa
      181 aagttggcac catgcccgcc atgccctcca gcggccccgg ggacaccagc agctctgctg
      241 cggagcggga ggaggaccga aaggacggag aggagcagga ggagccgcgt ggcaaggagg
      301 agcgccaaga gcccagcacc acggcacgga aggtggggcg gcctgggagg aagcgcaagc
      361 accccccggt ggaaagcggt gacacgccaa aggaccctgc ggtgatctcc aagtccccat
      421 ccatggccca ggactcaggc gcctcagagc tattacccaa tggggacttg gagaagcgga
      481 gtgagcccca gccagaggag gggagccctg ctggggggca gaagggcggg gccccagcag
      541 agggagaggg tgcagctgag accctgcctg aagcctcaag agcagtggaa aatggctgct
      601 gcacccccaa ggagggccga ggagcccctg cagaagcggg caaagaacag aaggagacca
      661 acatcgaatc catgaaaatg gagggctccc ggggccggct gcggggtggc ttgggctggg
      721 agtccagcct ccgtcagcgg cccatgccga ggctcacctt ccaggcgggg gacccctact
      781 acatcagcaa gcgcaagcgg gacgagtggc tggcacgctg gaaaagggag gctgagaaga
      841 aagccaaggt cattgcagga atgaatgctg tggaagaaaa ccaggggccc ggggagtctc
      901 agaaggtgga ggaggccagc cctcctgctg tgcagcagcc cactgacccc gcatccccca
      961 ctgtggctac cacgcctgag cccgtggggt ccgatgctgg ggacaagaat gccaccaaag
     1021 caggcgatga cgagccagag tacgaggacg gccggggctt tggcattggg gagctggtgt
     1081 gggggaaact gcggggcttc tcctggtggc caggccgcat tgtgtcttgg tggatgacgg
     1141 gccggagccg agcagctgaa ggcacccgct gggtcatgtg gttcggagac ggcaaattct
     1201 cagtggtgtg tgttgagaag ctgatgccgc tgagctcgtt ttgcagtgcg ttccaccagg
     1261 ccacgtacaa caagcagccc atgtaccgca aagccatcta cgaggtcctg caggtggcca
     1321 gcagccgcgc ggggaagctg ttcccggtgt gccacgacag cgatgagagt gacactgcca
     1381 aggccgtgga ggtgcagaac aagcccatga ttgaatgggc cctggggggc ttccagcctt
     1441 ctggccctaa gggcctggag ccaccagaag aagagaagaa tccctacaaa gaagtgtaca
     1501 cggacatgtg ggtggaacct gaggcagctg cctacgcacc acctccacca gccaaaaagc
     1561 cccggaagag cacagcggag aagcccaagg tcaaggagat tattgatgag cgcacaagag
     1621 agcggctggt gtacgaggtg cggcagaagt gccggaacat tgaggacatc tgcatctcct
     1681 gtgggagcct caatgttacc ctggaacacc ccctcttcgt tggaggaatg tgccaaaact
     1741 gcaagaactg ctttctggag tgtgcgtacc agtacgacga cgacggctac cagtcctact
     1801 gcaccatctg ctgtgggggc cgtgaggtgc tcatgtgcgg aaacaacaac tgctgcaggt
     1861 gcttttgcgt ggagtgtgtg gacctcttgg tggggccggg ggctgcccag gcagccatta
     1921 aggaagaccc ctggaactgc tacatgtgcg ggcacaaggg tacctacggg ctgctgcggc
     1981 ggcgagagga ctggccctcc cggctccaga tgttcttcgc taataaccac gaccaggaat
     2041 ttgaccctcc aaaggtttac ccacctgtcc cagctgagaa gaggaagccc atccgggtgc
     2101 tgtctctctt tgatggaatc gctacagggc tcctggtgct gaaggacttg ggcattcagg
     2161 tggaccgcta cattgcctcg gaggtgtgtg aggactccat cacggtgggc atggtgcggc
     2221 accaggggaa gatcatgtac gtcggggacg tccgcagcgt cacacagaag catatccagg
     2281 agtggggccc attcgatctg gtgattgggg gcagtccctg caatgacctc tccatcgtca
     2341 accctgctcg caagggcctc tacgagggca ctggccggct cttctttgag ttctaccgcc
     2401 tcctgcatga tgcgcggccc aaggagggag atgatcgccc cttcttctgg ctctttgaga
     2461 atgtggtggc catgggcgtt agtgacaaga gggacatctc gcgatttctc gagtccaacc
     2521 ctgtgatgat tgatgccaaa gaagtgtcag ctgcacacag ggcccgctac ttctggggta
     2581 accttcccgg tatgaacagg ccgttggcat ccactgtgaa tgataagctg gagctgcagg
     2641 agtgtctgga gcatggcagg atagccaagt tcagcaaagt gaggaccatt actacgaggt
     2701 caaactccat aaagcagggc aaagaccagc attttcctgt cttcatgaat gagaaagagg
     2761 acatcttatg gtgcactgaa atggaaaggg tatttggttt cccagtccac tatactgacg
     2821 tctccaacat gagccgcttg gcgaggcaga gactgctggg ccggtcatgg agcgtgccag
     2881 tcatccgcca cctcttcgct ccgctgaagg agtattttgt gtgtgtgtga ttgccaactt
     2941 tcttgtacaa agttggcatt ataagaaagc attgcttatc aatttgttgc aacgaacagg
     3001 tcactatcag tcaaaataaa atcattattt gccatccagc tgatatcccc tatagtgagt
     3061 cgtattacat ggtcatagct gtttcctggc agctctggcc cgtgtctcaa aatctctgat
     3121 gttacattgc acaagataaa aatatatcat catgcctcct ctagaccagc caggacagaa
     3181 atgcctcgac ttcgctgcta cccaaggttg ccgggtgacg cacaccgtgg aaacggatga
     3241 aggcacgaac ccagtggaca taagcctgtt cggttcgtaa gctgtaatgc aagtagcgta
     3301 tgcgctcacg caactggtcc agaaccttga ccgaacgcag cggtggtaac ggcgcagtgg
     3361 cggttttcat ggcttgttat gactgttttt ttggggtaca gtctatgcct cgggcatcca
     3421 agcagcaagc gcgttacgcc gtgggtcgat gtttgatgtt atggagcagc aacgatgtta
     3481 cgcagcaggg cagtcgccct aaaacaaagt taaacattat gagggaagcg gtgatcgccg
     3541 aagtatcgac tcaactatca gaggtagttg gcgtcatcga gcgccatctc gaaccgacgt
     3601 tgctggccgt acatttgtac ggctccgcag tggatggcgg cctgaagcca cacagtgata
     3661 ttgatttgct ggttacggtg accgtaaggc ttgatgaaac aacgcggcga gctttgatca
     3721 acgacctttt ggaaacttcg gcttcccctg gagagagcga gattctccgc gctgtagaag
     3781 tcaccattgt tgtgcacgac gacatcattc cgtggcgtta tccagctaag cgcgaactgc
     3841 aatttggaga atggcagcgc aatgacattc ttgcaggtat cttcgagcca gccacgatcg
     3901 acattgatct ggctatcttg ctgacaaaag caagagaaca tagcgttgcc ttggtaggtc
     3961 cagcggcgga ggaactcttt gatccggttc ctgaacagga tctatttgag gcgctaaatg
     4021 aaaccttaac gctatggaac tcgccgcccg actgggctgg cgatgagcga aatgtagtgc
     4081 ttacgttgtc ccgcatttgg tacagcgcag taaccggcaa aatcgcgccg aaggatgtcg
     4141 ctgccgactg ggcaatggag cgcctgccgg cccagtatca gcccgtcata cttgaagcta
     4201 gacaggctta tcttggacaa gaagaagatc gcttggcctc gcgcgcagat cagttggaag
     4261 aatttgtcca ctacgtgaaa ggcgagatca ccaaggtagt cggcaaataa ccctcgagcc
     4321 acccatgacc aaaatccctt aacgtgagtt acgcgtcgtt ccactgagcg tcagaccccg
     4381 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     4441 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     4501 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     4561 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     4621 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     4681 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     4741 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     4801 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     4861 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     4921 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     4981 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     5041 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     5101 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     5161 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     5221 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
     5281 atacgcgtac cgctagccag gaagagtttg tagaaacgca aaaaggccat ccgtcaggat
     5341 ggccttctgc ttagtttgat gcctggcagt ttatggcggg cgtcctgccc gccaccctcc
     5401 gggccgttgc ttcacaacgt tcaaatccgc tcccggcgga tttgtcctac tcaggagagc
     5461 gttcaccgac aaacaacaga taaaacgaaa ggcccagtct tccgactgag cctttcgttt
     5521 tatttgatgc ct
//
