LOCUS       sequence-164899-        4446 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 4446)
  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 4446)
  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_WT
FEATURES             Location/Qualifiers
     source          1..4446
                     /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(1846..1945)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1962..1981)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1963..1981)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1986..2002)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1986..2002)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2433..3224
                     /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      3317..3905
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3806..3825
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4059..4076
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4235..4262)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4354..4440)
                     /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 ggacaatatg tctattacga atacaccaac aagtaatgat gcctgtctga
      241 gcattgtgca tagtttgatg tgccatagac aaggtggaga gagtgaaaca tttgcaaaaa
      301 gagcaattga aagtttggta aagaagctga aggagaaaaa agatgaattg gattctttaa
      361 taacagctat aactacaaat ggagctcatc ctagtaaatg tgttaccata cagagaacat
      421 tggatgggag gcttcaggtg gctggtcgga aaggatttcc tcatgtgatc tatgcccgtc
      481 tctggaggtg gcctgatctt cacaaaaatg aactaaaaca tgttaaatat tgtcagtatg
      541 cgtttgactt aaaatgtgat agtgtctgtg tgaatccata tcactacgaa cgagttgtat
      601 cacctggaat tgatctctca ggattaacac tgcagagtaa tgctccatca agtatgatgg
      661 tgaaggatga atatgtgcat gactttgagg gacagccatc gttgtccact gaaggacatt
      721 caattcaaac catccagcat ccaccaagta atcgtgcatc gacagagaca tacagcaccc
      781 cagctctgtt agccccatct gagtctaatg ctaccagcac tgccaacttt cccaacattc
      841 ctgtggcttc cacaagtcag cctgccagta tactgggggg cagccatagt gaaggactgt
      901 tgcagatagc atcagggcct cagccaggac agcagcagaa tggatttact ggtcagccag
      961 ctacttacca tcataacagc actaccacct ggactggaag taggactgca ccatacacac
     1021 ctaatttgcc tcaccaccaa aacggccatc ttcagcacca cccgcctatg ccgccccatc
     1081 ccggacatta ctggcctgtt cacaatgagc ttgcattcca gcctcccatt tccaatcatc
     1141 ctgctcctga gtattggtgt tccattgctt actttgaaat ggatgttcag gtaggagaga
     1201 catttaaggt tccttcaagc tgccctattg ttactgttga tggatacgtg gacccttctg
     1261 gaggagatcg cttttgtttg ggtcaactct ccaatgtcca caggacagaa gccattgaga
     1321 gagcaaggtt gcacataggc aaaggtgtgc agttggaatg taaaggtgaa ggtgatgttt
     1381 gggtcaggtg ccttagtgac cacgcggtct ttgtacagag ttactactta gacagagaag
     1441 ctgggcgtgc acctggagat gctgttcata agatctaccc aagtgcatat ataaaggtct
     1501 ttgatttgcg tcagtgtcat cgacagatgc agcagcaggc ggctactgca caagctgcag
     1561 cagctgccca ggcagcagcc gtggcaggaa acatccctgg cccaggatca gtaggtggaa
     1621 tagctccagc tatcagtctg tcagctgctg ctggaattgg tgttgatgac cttcgtcgct
     1681 tatgcatact caggatgagt tttgtgaaag gctggggacc ggattaccca agacagagca
     1741 tcaaagaaac accttgctgg attgaaattc acttacaccg ggccctccag ctcctagacg
     1801 aagtacttca taccatgccg attgcagacc cacaaccttt agactgccca actttcttgt
     1861 acaaagttgg cattataaga aagcattgct tatcaatttg ttgcaacgaa caggtcacta
     1921 tcagtcaaaa taaaatcatt atttgccatc cagctgatat cccctatagt gagtcgtatt
     1981 acatggtcat agctgtttcc tggcagctct ggcccgtgtc tcaaaatctc tgatgttaca
     2041 ttgcacaaga taaaaatata tcatcatgcc tcctctagac cagccaggac agaaatgcct
     2101 cgacttcgct gctacccaag gttgccgggt gacgcacacc gtggaaacgg atgaaggcac
     2161 gaacccagtg gacataagcc tgttcggttc gtaagctgta atgcaagtag cgtatgcgct
     2221 cacgcaactg gtccagaacc ttgaccgaac gcagcggtgg taacggcgca gtggcggttt
     2281 tcatggcttg ttatgactgt ttttttgggg tacagtctat gcctcgggca tccaagcagc
     2341 aagcgcgtta cgccgtgggt cgatgtttga tgttatggag cagcaacgat gttacgcagc
     2401 agggcagtcg ccctaaaaca aagttaaaca ttatgaggga agcggtgatc gccgaagtat
     2461 cgactcaact atcagaggta gttggcgtca tcgagcgcca tctcgaaccg acgttgctgg
     2521 ccgtacattt gtacggctcc gcagtggatg gcggcctgaa gccacacagt gatattgatt
     2581 tgctggttac ggtgaccgta aggcttgatg aaacaacgcg gcgagctttg atcaacgacc
     2641 ttttggaaac ttcggcttcc cctggagaga gcgagattct ccgcgctgta gaagtcacca
     2701 ttgttgtgca cgacgacatc attccgtggc gttatccagc taagcgcgaa ctgcaatttg
     2761 gagaatggca gcgcaatgac attcttgcag gtatcttcga gccagccacg atcgacattg
     2821 atctggctat cttgctgaca aaagcaagag aacatagcgt tgccttggta ggtccagcgg
     2881 cggaggaact ctttgatccg gttcctgaac aggatctatt tgaggcgcta aatgaaacct
     2941 taacgctatg gaactcgccg cccgactggg ctggcgatga gcgaaatgta gtgcttacgt
     3001 tgtcccgcat ttggtacagc gcagtaaccg gcaaaatcgc gccgaaggat gtcgctgccg
     3061 actgggcaat ggagcgcctg ccggcccagt atcagcccgt catacttgaa gctagacagg
     3121 cttatcttgg acaagaagaa gatcgcttgg cctcgcgcgc agatcagttg gaagaatttg
     3181 tccactacgt gaaaggcgag atcaccaagg tagtcggcaa ataaccctcg agccacccat
     3241 gaccaaaatc ccttaacgtg agttacgcgt cgttccactg agcgtcagac cccgtagaaa
     3301 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     3361 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     3421 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     3481 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     3541 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     3601 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     3661 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     3721 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     3781 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     3841 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     3901 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     3961 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     4021 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     4081 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     4141 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatacgc
     4201 gtaccgctag ccaggaagag tttgtagaaa cgcaaaaagg ccatccgtca ggatggcctt
     4261 ctgcttagtt tgatgcctgg cagtttatgg cgggcgtcct gcccgccacc ctccgggccg
     4321 ttgcttcaca acgttcaaat ccgctcccgg cggatttgtc ctactcagga gagcgttcac
     4381 cgacaaacaa cagataaaac gaaaggccca gtcttccgac tgagcctttc gttttatttg
     4441 atgcct
//
