LOCUS       sequence-165291-        4065 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway Donor vector containing SMAD3, part of the Target 
            Accelerator Plasmid Collection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4065)
  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 4065)
  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_SMAD3_WT
FEATURES             Location/Qualifiers
     source          1..4065
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      40..628
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     529..548
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     782..799
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(958..985)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(1077..1163)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     1212..1234
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1226..1243
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1227..1243
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1259..1358
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(2634..2733)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2750..2769)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2751..2769)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2774..2790)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2774..2790)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3221..4012
                     /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"
ORIGIN
        1 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
       61 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
      121 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
      181 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
      241 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
      301 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
      361 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
      421 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
      481 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
      541 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
      601 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
      661 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
      721 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
      781 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
      841 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
      901 tgagcgcaac gcaattaata cgcgtaccgc tagccaggaa gagtttgtag aaacgcaaaa
      961 aggccatccg tcaggatggc cttctgctta gtttgatgcc tggcagttta tggcgggcgt
     1021 cctgcccgcc accctccggg ccgttgcttc acaacgttca aatccgctcc cggcggattt
     1081 gtcctactca ggagagcgtt caccgacaaa caacagataa aacgaaaggc ccagtcttcc
     1141 gactgagcct ttcgttttat ttgatgcctg gcagttccct actctcgcgt taacgctagc
     1201 atggatgttt tcccagtcac gacgttgtaa aacgacggcc agtcttaagc tcgggcccca
     1261 aataatgatt ttattttgac tgatagtgac ctgttcgttg caacaaattg atgagcaatg
     1321 cttttttata atgccaactt tgtacaaaaa agttggcatg tcgtccatcc tgcctttcac
     1381 tcccccgatc gtgaagcgcc tgctgggctg gaagaagggc gagcagaacg ggcaggagga
     1441 gaaatggtgc gagaaggcgg tcaagagcct ggtcaagaaa ctcaagaaga cggggcagct
     1501 ggacgagctg gagaaggcca tcaccacgca gaacgtcaac accaagtgca tcaccatccc
     1561 caggtccctg gatggccggt tgcaggtgtc ccatcggaag gggctccctc atgtcatcta
     1621 ctgccgcctg tggcgatggc cagacctgca cagccaccac gagctgcggg ccatggagct
     1681 gtgtgagttc gccttcaata tgaagaagga cgaggtctgc gtgaatccct accactacca
     1741 gagagtagag acaccagttc tacctcctgt gttggtgcca cgccacacag agatcccggc
     1801 cgagttcccc ccactggacg actacagcca ttccatcccc gaaaacacta acttccccgc
     1861 aggcatcgag ccccagagca atattccaga gaccccaccc cctggctacc tgagtgaaga
     1921 tggagaaacc agtgaccacc agatgaacca cagcatggac gcaggttctc caaacctatc
     1981 cccgaatccg atgtccccag cacataataa cttggacctg cagccagtta cctactgcga
     2041 gccggccttc tggtgctcca tctcctacta cgagctgaac cagcgcgtcg gggagacatt
     2101 ccacgcctcg cagccatcca tgactgtgga tggcttcacc gacccctcca attcggagcg
     2161 cttctgccta gggctgctct ccaatgtcaa caggaatgca gcagtggagc tgacacggag
     2221 acacatcgga agaggcgtgc ggctctacta catcggaggg gaggtcttcg cagagtgcct
     2281 cagtgacagc gctatttttg tccagtctcc caactgtaac cagcgctatg gctggcaccc
     2341 ggccaccgtc tgcaagatcc caccaggatg caacctgaag atcttcaaca accaggagtt
     2401 cgctgccctc ctggcccagt cggtcaacca gggctttgag gctgtctacc agttgacccg
     2461 aatgtgcacc atccgcatga gcttcgtcaa aggctgggga gcggagtaca ggagacagac
     2521 tgtgaccagt accccctgct ggattgagct gcacctgaat gggcctttgc agtggcttga
     2581 caaggtcctc acccagatgg gctccccaag catccgctgt tccagtgtgt cttacccaac
     2641 tttcttgtac aaagttggca ttataagaaa gcattgctta tcaatttgtt gcaacgaaca
     2701 ggtcactatc agtcaaaata aaatcattat ttgccatcca gctgatatcc cctatagtga
     2761 gtcgtattac atggtcatag ctgtttcctg gcagctctgg cccgtgtctc aaaatctctg
     2821 atgttacatt gcacaagata aaaatatatc atcatgcctc ctctagacca gccaggacag
     2881 aaatgcctcg acttcgctgc tacccaaggt tgccgggtga cgcacaccgt ggaaacggat
     2941 gaaggcacga acccagtgga cataagcctg ttcggttcgt aagctgtaat gcaagtagcg
     3001 tatgcgctca cgcaactggt ccagaacctt gaccgaacgc agcggtggta acggcgcagt
     3061 ggcggttttc atggcttgtt atgactgttt ttttggggta cagtctatgc ctcgggcatc
     3121 caagcagcaa gcgcgttacg ccgtgggtcg atgtttgatg ttatggagca gcaacgatgt
     3181 tacgcagcag ggcagtcgcc ctaaaacaaa gttaaacatt atgagggaag cggtgatcgc
     3241 cgaagtatcg actcaactat cagaggtagt tggcgtcatc gagcgccatc tcgaaccgac
     3301 gttgctggcc gtacatttgt acggctccgc agtggatggc ggcctgaagc cacacagtga
     3361 tattgatttg ctggttacgg tgaccgtaag gcttgatgaa acaacgcggc gagctttgat
     3421 caacgacctt ttggaaactt cggcttcccc tggagagagc gagattctcc gcgctgtaga
     3481 agtcaccatt gttgtgcacg acgacatcat tccgtggcgt tatccagcta agcgcgaact
     3541 gcaatttgga gaatggcagc gcaatgacat tcttgcaggt atcttcgagc cagccacgat
     3601 cgacattgat ctggctatct tgctgacaaa agcaagagaa catagcgttg ccttggtagg
     3661 tccagcggcg gaggaactct ttgatccggt tcctgaacag gatctatttg aggcgctaaa
     3721 tgaaacctta acgctatgga actcgccgcc cgactgggct ggcgatgagc gaaatgtagt
     3781 gcttacgttg tcccgcattt ggtacagcgc agtaaccggc aaaatcgcgc cgaaggatgt
     3841 cgctgccgac tgggcaatgg agcgcctgcc ggcccagtat cagcccgtca tacttgaagc
     3901 tagacaggct tatcttggac aagaagaaga tcgcttggcc tcgcgcgcag atcagttgga
     3961 agaatttgtc cactacgtga aaggcgagat caccaaggta gtcggcaaat aaccctcgag
     4021 ccacccatga ccaaaatccc ttaacgtgag ttacgcgtcg ttcca
//
