LOCUS       sequence-164902-        4185 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway Donor vector containing SMAD5 , part of the Target 
            Accelerator Plasmid Collection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4185)
  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 4185)
  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_SMAD5_WT
FEATURES             Location/Qualifiers
     source          1..4185
                     /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"
     protein_bind    complement(1585..1684)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1701..1720)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1702..1720)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1725..1741)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1725..1741)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2172..2963
                     /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      3056..3644
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3545..3564
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3798..3815
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3974..4001)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4093..4179)
                     /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 aagttggcat gacgtcaatg gccagcttgt tttcttttac tagtccagca gtaaagcgat
      241 tgttgggctg gaaacaaggt gatgaggagg agaaatgggc agaaaaggca gttgatgctt
      301 tggtgaagaa actaaaaaag aaaaagggtg ccatggagga actggagaaa gccttgagca
      361 gtccaggaca gccgagtaaa tgtgtcacta ttcccagatc tttagatgga cgcctgcagg
      421 tttctcacag aaaaggctta ccccatgtta tatattgtcg tgtttggcgc tggccggatt
      481 tgcagagtca tcatgagcta aagccgttgg atatttgtga atttcctttt ggatctaagc
      541 aaaaagaagt ttgtatcaac ccataccact ataagagagt ggagagtcca gtcttacctc
      601 cagtattagt gcctcgtcat aatgaattca atccacaaca cagccttctg gttcagttta
      661 ggaacctgag ccacaatgaa ccacacatgc cacaaaatgc cacgtttcca gattctttcc
      721 accagcccaa caacactcct tttcccttat ctccaaacag cccttatccc ccttctcctg
      781 ctagcagcac atatcccaac tccccagcaa gttctggacc aggaagtcca tttcagctcc
      841 cagctgatac gcctcctcct gcctatatgc cacctgatga tcagatgggt caagataatt
      901 cccagcctat ggatacaagc aataatatga ttcctcagat tatgcccagt atatccagca
      961 gggatgttca gcctgttgcc tatgaagagc ctaaacattg gtgttcaata gtctactatg
     1021 aattaaacaa tcgtgttgga gaagcttttc atgcatcttc tactagtgtg ttagtagatg
     1081 gattcacaga tccttcaaat aacaaaagta gattctgctt gggtttgttg tcaaatgtta
     1141 atcgtaattc gacaattgaa aacactaggc gacatattgg aaaaggtgtt catctgtact
     1201 atgttggtgg agaggtgtat gcggaatgcc tcagtgacag cagcatattt gtacagagta
     1261 ggaactgcaa ctttcatcat ggctttcatc ccaccactgt ctgtaagatt cccagcagct
     1321 gcagcctcaa aatttttaac aatcaggagt ttgctcagct tctggctcaa tctgtcaacc
     1381 atgggtttga ggcagtatat gagctcacca aaatgtgtac cattcggatg agttttgtca
     1441 agggttgggg agcagaatat caccggcagg atgtaaccag caccccatgt tggattgaga
     1501 ttcatcttca tgggcctctt cagtggctgg ataaagtcct tactcagatg ggctcccctc
     1561 tgaaccccat atcttctgtt tcatacccaa ctttcttgta caaagttggc attataagaa
     1621 agcattgctt atcaatttgt tgcaacgaac aggtcactat cagtcaaaat aaaatcatta
     1681 tttgccatcc agctgatatc ccctatagtg agtcgtatta catggtcata gctgtttcct
     1741 ggcagctctg gcccgtgtct caaaatctct gatgttacat tgcacaagat aaaaatatat
     1801 catcatgcct cctctagacc agccaggaca gaaatgcctc gacttcgctg ctacccaagg
     1861 ttgccgggtg acgcacaccg tggaaacgga tgaaggcacg aacccagtgg acataagcct
     1921 gttcggttcg taagctgtaa tgcaagtagc gtatgcgctc acgcaactgg tccagaacct
     1981 tgaccgaacg cagcggtggt aacggcgcag tggcggtttt catggcttgt tatgactgtt
     2041 tttttggggt acagtctatg cctcgggcat ccaagcagca agcgcgttac gccgtgggtc
     2101 gatgtttgat gttatggagc agcaacgatg ttacgcagca gggcagtcgc cctaaaacaa
     2161 agttaaacat tatgagggaa gcggtgatcg ccgaagtatc gactcaacta tcagaggtag
     2221 ttggcgtcat cgagcgccat ctcgaaccga cgttgctggc cgtacatttg tacggctccg
     2281 cagtggatgg cggcctgaag ccacacagtg atattgattt gctggttacg gtgaccgtaa
     2341 ggcttgatga aacaacgcgg cgagctttga tcaacgacct tttggaaact tcggcttccc
     2401 ctggagagag cgagattctc cgcgctgtag aagtcaccat tgttgtgcac gacgacatca
     2461 ttccgtggcg ttatccagct aagcgcgaac tgcaatttgg agaatggcag cgcaatgaca
     2521 ttcttgcagg tatcttcgag ccagccacga tcgacattga tctggctatc ttgctgacaa
     2581 aagcaagaga acatagcgtt gccttggtag gtccagcggc ggaggaactc tttgatccgg
     2641 ttcctgaaca ggatctattt gaggcgctaa atgaaacctt aacgctatgg aactcgccgc
     2701 ccgactgggc tggcgatgag cgaaatgtag tgcttacgtt gtcccgcatt tggtacagcg
     2761 cagtaaccgg caaaatcgcg ccgaaggatg tcgctgccga ctgggcaatg gagcgcctgc
     2821 cggcccagta tcagcccgtc atacttgaag ctagacaggc ttatcttgga caagaagaag
     2881 atcgcttggc ctcgcgcgca gatcagttgg aagaatttgt ccactacgtg aaaggcgaga
     2941 tcaccaaggt agtcggcaaa taaccctcga gccacccatg accaaaatcc cttaacgtga
     3001 gttacgcgtc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     3061 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     3121 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     3181 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     3241 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     3301 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     3361 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     3421 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     3481 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     3541 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     3601 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     3661 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     3721 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     3781 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca
     3841 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
     3901 actggaaagc gggcagtgag cgcaacgcaa ttaatacgcg taccgctagc caggaagagt
     3961 ttgtagaaac gcaaaaaggc catccgtcag gatggccttc tgcttagttt gatgcctggc
     4021 agtttatggc gggcgtcctg cccgccaccc tccgggccgt tgcttcacaa cgttcaaatc
     4081 cgctcccggc ggatttgtcc tactcaggag agcgttcacc gacaaacaac agataaaacg
     4141 aaaggcccag tcttccgact gagcctttcg ttttatttga tgcct
//
