LOCUS       sequence-168878-        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_WT
FEATURES             Location/Qualifiers
     source          1..5532
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             671..3409
                     /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
                     VHYTDVSNMSRLARQRLLGRSWSVPVIRHLFAPLKEYFACV"
     protein_bind    complement(3411..3510)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3527..3546)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3528..3546)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3551..3567)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3551..3567)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3998..4789
                     /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      4882..5470
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5371..5390
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agttggcacc atgcccgcca tgccctccag cggccccggg gacaccagca gctctgctgc
      721 ggagcgggag gaggaccgaa aggacggaga ggagcaggag gagccgcgtg gcaaggagga
      781 gcgccaagag cccagcacca cggcacggaa ggtggggcgg cctgggagga agcgcaagca
      841 ccccccggtg gaaagcggtg acacgccaaa ggaccctgcg gtgatctcca agtccccatc
      901 catggcccag gactcaggcg cctcagagct attacccaat ggggacttgg agaagcggag
      961 tgagccccag ccagaggagg ggagccctgc tggggggcag aagggcgggg ccccagcaga
     1021 gggagagggt gcagctgaga ccctgcctga agcctcaaga gcagtggaaa atggctgctg
     1081 cacccccaag gagggccgag gagcccctgc agaagcgggc aaagaacaga aggagaccaa
     1141 catcgaatcc atgaaaatgg agggctcccg gggccggctg cggggtggct tgggctggga
     1201 gtccagcctc cgtcagcggc ccatgccgag gctcaccttc caggcggggg acccctacta
     1261 catcagcaag cgcaagcggg acgagtggct ggcacgctgg aaaagggagg ctgagaagaa
     1321 agccaaggtc attgcaggaa tgaatgctgt ggaagaaaac caggggcccg gggagtctca
     1381 gaaggtggag gaggccagcc ctcctgctgt gcagcagccc actgaccccg catcccccac
     1441 tgtggctacc acgcctgagc ccgtggggtc cgatgctggg gacaagaatg ccaccaaagc
     1501 aggcgatgac gagccagagt acgaggacgg ccggggcttt ggcattgggg agctggtgtg
     1561 ggggaaactg cggggcttct cctggtggcc aggccgcatt gtgtcttggt ggatgacggg
     1621 ccggagccga gcagctgaag gcacccgctg ggtcatgtgg ttcggagacg gcaaattctc
     1681 agtggtgtgt gttgagaagc tgatgccgct gagctcgttt tgcagtgcgt tccaccaggc
     1741 cacgtacaac aagcagccca tgtaccgcaa agccatctac gaggtcctgc aggtggccag
     1801 cagccgcgcg gggaagctgt tcccggtgtg ccacgacagc gatgagagtg acactgccaa
     1861 ggccgtggag gtgcagaaca agcccatgat tgaatgggcc ctggggggct tccagccttc
     1921 tggccctaag ggcctggagc caccagaaga agagaagaat ccctacaaag aagtgtacac
     1981 ggacatgtgg gtggaacctg aggcagctgc ctacgcacca cctccaccag ccaaaaagcc
     2041 ccggaagagc acagcggaga agcccaaggt caaggagatt attgatgagc gcacaagaga
     2101 gcggctggtg tacgaggtgc ggcagaagtg ccggaacatt gaggacatct gcatctcctg
     2161 tgggagcctc aatgttaccc tggaacaccc cctcttcgtt ggaggaatgt gccaaaactg
     2221 caagaactgc tttctggagt gtgcgtacca gtacgacgac gacggctacc agtcctactg
     2281 caccatctgc tgtgggggcc gtgaggtgct catgtgcgga aacaacaact gctgcaggtg
     2341 cttttgcgtg gagtgtgtgg acctcttggt ggggccgggg gctgcccagg cagccattaa
     2401 ggaagacccc tggaactgct acatgtgcgg gcacaagggt acctacgggc tgctgcggcg
     2461 gcgagaggac tggccctccc ggctccagat gttcttcgct aataaccacg accaggaatt
     2521 tgaccctcca aaggtttacc cacctgtccc agctgagaag aggaagccca tccgggtgct
     2581 gtctctcttt gatggaatcg ctacagggct cctggtgctg aaggacttgg gcattcaggt
     2641 ggaccgctac attgcctcgg aggtgtgtga ggactccatc acggtgggca tggtgcggca
     2701 ccaggggaag atcatgtacg tcggggacgt ccgcagcgtc acacagaagc atatccagga
     2761 gtggggccca ttcgatctgg tgattggggg cagtccctgc aatgacctct ccatcgtcaa
     2821 ccctgctcgc aagggcctct acgagggcac tggccggctc ttctttgagt tctaccgcct
     2881 cctgcatgat gcgcggccca aggagggaga tgatcgcccc ttcttctggc tctttgagaa
     2941 tgtggtggcc atgggcgtta gtgacaagag ggacatctcg cgatttctcg agtccaaccc
     3001 tgtgatgatt gatgccaaag aagtgtcagc tgcacacagg gcccgctact tctggggtaa
     3061 ccttcccggt atgaacaggc cgttggcatc cactgtgaat gataagctgg agctgcagga
     3121 gtgtctggag catggcagga tagccaagtt cagcaaagtg aggaccatta ctacgaggtc
     3181 aaactccata aagcagggca aagaccagca ttttcctgtc ttcatgaatg agaaagagga
     3241 catcttatgg tgcactgaaa tggaaagggt atttggtttc ccagtccact atactgacgt
     3301 ctccaacatg agccgcttgg cgaggcagag actgctgggc cggtcatgga gcgtgccagt
     3361 catccgccac ctcttcgctc cgctgaagga gtattttgcg tgtgtgtgat tgccaacttt
     3421 cttgtacaaa gttggcatta taagaaagca ttgcttatca atttgttgca acgaacaggt
     3481 cactatcagt caaaataaaa tcattatttg ccatccagct gatatcccct atagtgagtc
     3541 gtattacatg gtcatagctg tttcctggca gctctggccc gtgtctcaaa atctctgatg
     3601 ttacattgca caagataaaa atatatcatc atgcctcctc tagaccagcc aggacagaaa
     3661 tgcctcgact tcgctgctac ccaaggttgc cgggtgacgc acaccgtgga aacggatgaa
     3721 ggcacgaacc cagtggacat aagcctgttc ggttcgtaag ctgtaatgca agtagcgtat
     3781 gcgctcacgc aactggtcca gaaccttgac cgaacgcagc ggtggtaacg gcgcagtggc
     3841 ggttttcatg gcttgttatg actgtttttt tggggtacag tctatgcctc gggcatccaa
     3901 gcagcaagcg cgttacgccg tgggtcgatg tttgatgtta tggagcagca acgatgttac
     3961 gcagcagggc agtcgcccta aaacaaagtt aaacattatg agggaagcgg tgatcgccga
     4021 agtatcgact caactatcag aggtagttgg cgtcatcgag cgccatctcg aaccgacgtt
     4081 gctggccgta catttgtacg gctccgcagt ggatggcggc ctgaagccac acagtgatat
     4141 tgatttgctg gttacggtga ccgtaaggct tgatgaaaca acgcggcgag ctttgatcaa
     4201 cgaccttttg gaaacttcgg cttcccctgg agagagcgag attctccgcg ctgtagaagt
     4261 caccattgtt gtgcacgacg acatcattcc gtggcgttat ccagctaagc gcgaactgca
     4321 atttggagaa tggcagcgca atgacattct tgcaggtatc ttcgagccag ccacgatcga
     4381 cattgatctg gctatcttgc tgacaaaagc aagagaacat agcgttgcct tggtaggtcc
     4441 agcggcggag gaactctttg atccggttcc tgaacaggat ctatttgagg cgctaaatga
     4501 aaccttaacg ctatggaact cgccgcccga ctgggctggc gatgagcgaa atgtagtgct
     4561 tacgttgtcc cgcatttggt acagcgcagt aaccggcaaa atcgcgccga aggatgtcgc
     4621 tgccgactgg gcaatggagc gcctgccggc ccagtatcag cccgtcatac ttgaagctag
     4681 acaggcttat cttggacaag aagaagatcg cttggcctcg cgcgcagatc agttggaaga
     4741 atttgtccac tacgtgaaag gcgagatcac caaggtagtc ggcaaataac cctcgagcca
     4801 cccatgacca aaatccctta acgtgagtta cgcgtcgttc cactgagcgt cagaccccgt
     4861 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     4921 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     4981 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     5041 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     5101 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     5161 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     5221 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     5281 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     5341 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     5401 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     5461 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     5521 tgctcacatg tt
//
