LOCUS       sequence-168327-        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.K519N
FEATURES             Location/Qualifiers
     source          1..4449
                     /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"
     protein_bind    complement(2328..2427)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2444..2463)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2445..2463)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2468..2484)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2468..2484)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2915..3706
                     /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      3799..4387
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4288..4307
                     /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 agttggcatg gacaatatgt ctattacgaa tacaccaaca agtaatgatg cctgtctgag
      721 cattgtgcat agtttgatgt gccatagaca aggtggagag agtgaaacat ttgcaaaaag
      781 agcaattgaa agtttggtaa agaagctgaa ggagaaaaaa gatgaattgg attctttaat
      841 aacagctata actacaaatg gagctcatcc tagtaaatgt gttaccatac agagaacatt
      901 ggatgggagg cttcaggtgg ctggtcggaa aggatttcct catgtgatct atgcccgtct
      961 ctggaggtgg cctgatcttc acaaaaatga actaaaacat gttaaatatt gtcagtatgc
     1021 gtttgactta aaatgtgata gtgtctgtgt gaatccatat cactacgaac gagttgtatc
     1081 acctggaatt gatctctcag gattaacact gcagagtaat gctccatcaa gtatgatggt
     1141 gaaggatgaa tatgtgcatg actttgaggg acagccatcg ttgtccactg aaggacattc
     1201 aattcaaacc atccagcatc caccaagtaa tcgtgcatcg acagagacat acagcacccc
     1261 agctctgtta gccccatctg agtctaatgc taccagcact gccaactttc ccaacattcc
     1321 tgtggcttcc acaagtcagc ctgccagtat actggggggc agccatagtg aaggactgtt
     1381 gcagatagca tcagggcctc agccaggaca gcagcagaat ggatttactg gtcagccagc
     1441 tacttaccat cataacagca ctaccacctg gactggaagt aggactgcac catacacacc
     1501 taatttgcct caccaccaaa acggccatct tcagcaccac ccgcctatgc cgccccatcc
     1561 cggacattac tggcctgttc acaatgagct tgcattccag cctcccattt ccaatcatcc
     1621 tgctcctgag tattggtgtt ccattgctta ctttgaaatg gatgttcagg taggagagac
     1681 atttaaggtt ccttcaagct gccctattgt tactgttgat ggatacgtgg acccttctgg
     1741 aggagatcgc ttttgtttgg gtcaactctc caatgtccac aggacagaag ccattgagag
     1801 agcaaggttg cacataggca aaggtgtgca gttggaatgt aaaggtgaag gtgatgtttg
     1861 ggtcaggtgc cttagtgacc acgcggtctt tgtacagagt tactacttag acagagaagc
     1921 tgggcgtgca cctggagatg ctgttcataa gatctaccca agtgcatata taaaggtctt
     1981 tgatttgcgt cagtgtcatc gacagatgca gcagcaggcg gctactgcac aagctgcagc
     2041 agctgcccag gcagcagccg tggcaggaaa catccctggc ccaggatcag taggtggaat
     2101 agctccagct atcagtctgt cagctgctgc tggaattggt gttgatgacc ttcgtcgctt
     2161 atgcatactc aggatgagtt ttgtgaaagg ctggggaccg gattacccaa gacagagcat
     2221 caacgaaaca ccttgctgga ttgaaattca cttacaccgg gccctccagc tcctagacga
     2281 agtacttcat accatgccga ttgcagaccc acaaccttta gactgattgc caactttctt
     2341 gtacaaagtt ggcattataa gaaagcattg cttatcaatt tgttgcaacg aacaggtcac
     2401 tatcagtcaa aataaaatca ttatttgcca tccagctgat atcccctata gtgagtcgta
     2461 ttacatggtc atagctgttt cctggcagct ctggcccgtg tctcaaaatc tctgatgtta
     2521 cattgcacaa gataaaaata tatcatcatg cctcctctag accagccagg acagaaatgc
     2581 ctcgacttcg ctgctaccca aggttgccgg gtgacgcaca ccgtggaaac ggatgaaggc
     2641 acgaacccag tggacataag cctgttcggt tcgtaagctg taatgcaagt agcgtatgcg
     2701 ctcacgcaac tggtccagaa ccttgaccga acgcagcggt ggtaacggcg cagtggcggt
     2761 tttcatggct tgttatgact gtttttttgg ggtacagtct atgcctcggg catccaagca
     2821 gcaagcgcgt tacgccgtgg gtcgatgttt gatgttatgg agcagcaacg atgttacgca
     2881 gcagggcagt cgccctaaaa caaagttaaa cattatgagg gaagcggtga tcgccgaagt
     2941 atcgactcaa ctatcagagg tagttggcgt catcgagcgc catctcgaac cgacgttgct
     3001 ggccgtacat ttgtacggct ccgcagtgga tggcggcctg aagccacaca gtgatattga
     3061 tttgctggtt acggtgaccg taaggcttga tgaaacaacg cggcgagctt tgatcaacga
     3121 ccttttggaa acttcggctt cccctggaga gagcgagatt ctccgcgctg tagaagtcac
     3181 cattgttgtg cacgacgaca tcattccgtg gcgttatcca gctaagcgcg aactgcaatt
     3241 tggagaatgg cagcgcaatg acattcttgc aggtatcttc gagccagcca cgatcgacat
     3301 tgatctggct atcttgctga caaaagcaag agaacatagc gttgccttgg taggtccagc
     3361 ggcggaggaa ctctttgatc cggttcctga acaggatcta tttgaggcgc taaatgaaac
     3421 cttaacgcta tggaactcgc cgcccgactg ggctggcgat gagcgaaatg tagtgcttac
     3481 gttgtcccgc atttggtaca gcgcagtaac cggcaaaatc gcgccgaagg atgtcgctgc
     3541 cgactgggca atggagcgcc tgccggccca gtatcagccc gtcatacttg aagctagaca
     3601 ggcttatctt ggacaagaag aagatcgctt ggcctcgcgc gcagatcagt tggaagaatt
     3661 tgtccactac gtgaaaggcg agatcaccaa ggtagtcggc aaataaccct cgagccaccc
     3721 atgaccaaaa tcccttaacg tgagttacgc gtcgttccac tgagcgtcag accccgtaga
     3781 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     3841 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     3901 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     3961 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     4021 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     4081 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     4141 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     4201 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     4261 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     4321 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     4381 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     4441 tcacatgtt
//
