LOCUS       sequence-168267-        4449 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway Donor vector containing SMAD4 , part of the Target 
            Accelerator Plasmid Collection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4449)
  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 4449)
  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_SMAD4_p.R97H
FEATURES             Location/Qualifiers
     source          1..4449
                     /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(1849..1948)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1965..1984)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1966..1984)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1989..2005)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1989..2005)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2436..3227
                     /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      3320..3908
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3809..3828
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4062..4079
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4238..4265)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4357..4443)
                     /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 aatgacaact ttgtacaaaa
      181 aagttggcac catggacaat atgtctatta cgaatacacc aacaagtaat gatgcctgtc
      241 tgagcattgt gcatagtttg atgtgccata gacaaggtgg agagagtgaa acatttgcaa
      301 aaagagcaat tgaaagtttg gtaaagaagc tgaaggagaa aaaagatgaa ttggattctt
      361 taataacagc tataactaca aatggagctc atcctagtaa atgtgttacc atacagagaa
      421 cattggatgg gaggcttcag gtggctggtc ggaaaggatt tcctcatgtg atctatgccc
      481 atctctggag gtggcctgat cttcacaaaa atgaactaaa acatgttaaa tattgtcagt
      541 atgcgtttga cttaaaatgt gatagtgtct gtgtgaatcc atatcactac gaacgagttg
      601 tatcacctgg aattgatctc tcaggattaa cactgcagag taatgctcca tcaagtatga
      661 tggtgaagga tgaatatgtg catgactttg agggacagcc atcgttgtcc actgaaggac
      721 attcaattca aaccatccag catccaccaa gtaatcgtgc atcgacagag acatacagca
      781 ccccagctct gttagcccca tctgagtcta atgctaccag cactgccaac tttcccaaca
      841 ttcctgtggc ttccacaagt cagcctgcca gtatactggg gggcagccat agtgaaggac
      901 tgttgcagat agcatcaggg cctcagccag gacagcagca gaatggattt actggtcagc
      961 cagctactta ccatcataac agcactacca cctggactgg aagtaggact gcaccataca
     1021 cacctaattt gcctcaccac caaaacggcc atcttcagca ccacccgcct atgccgcccc
     1081 atcccggaca ttactggcct gttcacaatg agcttgcatt ccagcctccc atttccaatc
     1141 atcctgctcc tgagtattgg tgttccattg cttactttga aatggatgtt caggtaggag
     1201 agacatttaa ggttccttca agctgcccta ttgttactgt tgatggatac gtggaccctt
     1261 ctggaggaga tcgcttttgt ttgggtcaac tctccaatgt ccacaggaca gaagccattg
     1321 agagagcaag gttgcacata ggcaaaggtg tgcagttgga atgtaaaggt gaaggtgatg
     1381 tttgggtcag gtgccttagt gaccacgcgg tctttgtaca gagttactac ttagacagag
     1441 aagctgggcg tgcacctgga gatgctgttc ataagatcta cccaagtgca tatataaagg
     1501 tctttgattt gcgtcagtgt catcgacaga tgcagcagca ggcggctact gcacaagctg
     1561 cagcagctgc ccaggcagca gccgtggcag gaaacatccc tggcccagga tcagtaggtg
     1621 gaatagctcc agctatcagt ctgtcagctg ctgctggaat tggtgttgat gaccttcgtc
     1681 gcttatgcat actcaggatg agttttgtga aaggctgggg accggattac ccaagacaga
     1741 gcatcaaaga aacaccttgc tggattgaaa ttcacttaca ccgggccctc cagctcctag
     1801 acgaagtact tcataccatg ccgattgcag acccacaacc tttagactgc ccaactttct
     1861 tgtacaaagt tggcattata agaaagcatt gcttatcaat ttgttgcaac gaacaggtca
     1921 ctatcagtca aaataaaatc attatttgcc atccagctga tatcccctat agtgagtcgt
     1981 attacatggt catagctgtt tcctggcagc tctggcccgt gtctcaaaat ctctgatgtt
     2041 acattgcaca agataaaaat atatcatcat gcctcctcta gaccagccag gacagaaatg
     2101 cctcgacttc gctgctaccc aaggttgccg ggtgacgcac accgtggaaa cggatgaagg
     2161 cacgaaccca gtggacataa gcctgttcgg ttcgtaagct gtaatgcaag tagcgtatgc
     2221 gctcacgcaa ctggtccaga accttgaccg aacgcagcgg tggtaacggc gcagtggcgg
     2281 ttttcatggc ttgttatgac tgtttttttg gggtacagtc tatgcctcgg gcatccaagc
     2341 agcaagcgcg ttacgccgtg ggtcgatgtt tgatgttatg gagcagcaac gatgttacgc
     2401 agcagggcag tcgccctaaa acaaagttaa acattatgag ggaagcggtg atcgccgaag
     2461 tatcgactca actatcagag gtagttggcg tcatcgagcg ccatctcgaa ccgacgttgc
     2521 tggccgtaca tttgtacggc tccgcagtgg atggcggcct gaagccacac agtgatattg
     2581 atttgctggt tacggtgacc gtaaggcttg atgaaacaac gcggcgagct ttgatcaacg
     2641 accttttgga aacttcggct tcccctggag agagcgagat tctccgcgct gtagaagtca
     2701 ccattgttgt gcacgacgac atcattccgt ggcgttatcc agctaagcgc gaactgcaat
     2761 ttggagaatg gcagcgcaat gacattcttg caggtatctt cgagccagcc acgatcgaca
     2821 ttgatctggc tatcttgctg acaaaagcaa gagaacatag cgttgccttg gtaggtccag
     2881 cggcggagga actctttgat ccggttcctg aacaggatct atttgaggcg ctaaatgaaa
     2941 ccttaacgct atggaactcg ccgcccgact gggctggcga tgagcgaaat gtagtgctta
     3001 cgttgtcccg catttggtac agcgcagtaa ccggcaaaat cgcgccgaag gatgtcgctg
     3061 ccgactgggc aatggagcgc ctgccggccc agtatcagcc cgtcatactt gaagctagac
     3121 aggcttatct tggacaagaa gaagatcgct tggcctcgcg cgcagatcag ttggaagaat
     3181 ttgtccacta cgtgaaaggc gagatcacca aggtagtcgg caaataaccc tcgagccacc
     3241 catgaccaaa atcccttaac gtgagttacg cgtcgttcca ctgagcgtca gaccccgtag
     3301 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     3361 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     3421 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     3481 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     3541 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     3601 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     3661 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     3721 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     3781 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     3841 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     3901 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     3961 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     4021 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     4081 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     4141 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata
     4201 cgcgtaccgc tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc
     4261 cttctgctta gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg
     4321 ccgttgcttc acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt
     4381 caccgacaaa caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat
     4441 ttgatgcct
//
