LOCUS       sequence-166738-        4452 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 4452)
  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 4452)
  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.R361P
FEATURES             Location/Qualifiers
     source          1..4452
                     /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(2331..2430)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2447..2466)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2448..2466)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2471..2487)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2471..2487)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2918..3709
                     /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      3802..4390
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4291..4310
                     /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 agttggcacc atggacaata tgtctattac gaatacacca acaagtaatg atgcctgtct
      721 gagcattgtg catagtttga tgtgccatag acaaggtgga gagagtgaaa catttgcaaa
      781 aagagcaatt gaaagtttgg taaagaagct gaaggagaaa aaagatgaat tggattcttt
      841 aataacagct ataactacaa atggagctca tcctagtaaa tgtgttacca tacagagaac
      901 attggatggg aggcttcagg tggctggtcg gaaaggattt cctcatgtga tctatgcccg
      961 tctctggagg tggcctgatc ttcacaaaaa tgaactaaaa catgttaaat attgtcagta
     1021 tgcgtttgac ttaaaatgtg atagtgtctg tgtgaatcca tatcactacg aacgagttgt
     1081 atcacctgga attgatctct caggattaac actgcagagt aatgctccat caagtatgat
     1141 ggtgaaggat gaatatgtgc atgactttga gggacagcca tcgttgtcca ctgaaggaca
     1201 ttcaattcaa accatccagc atccaccaag taatcgtgca tcgacagaga catacagcac
     1261 cccagctctg ttagccccat ctgagtctaa tgctaccagc actgccaact ttcccaacat
     1321 tcctgtggct tccacaagtc agcctgccag tatactgggg ggcagccata gtgaaggact
     1381 gttgcagata gcatcagggc ctcagccagg acagcagcag aatggattta ctggtcagcc
     1441 agctacttac catcataaca gcactaccac ctggactgga agtaggactg caccatacac
     1501 acctaatttg cctcaccacc aaaacggcca tcttcagcac cacccgccta tgccgcccca
     1561 tcccggacat tactggcctg ttcacaatga gcttgcattc cagcctccca tttccaatca
     1621 tcctgctcct gagtattggt gttccattgc ttactttgaa atggatgttc aggtaggaga
     1681 gacatttaag gttccttcaa gctgccctat tgttactgtt gatggatacg tggacccttc
     1741 tggaggagat cccttttgtt tgggtcaact ctccaatgtc cacaggacag aagccattga
     1801 gagagcaagg ttgcacatag gcaaaggtgt gcagttggaa tgtaaaggtg aaggtgatgt
     1861 ttgggtcagg tgccttagtg accacgcggt ctttgtacag agttactact tagacagaga
     1921 agctgggcgt gcacctggag atgctgttca taagatctac ccaagtgcat atataaaggt
     1981 ctttgatttg cgtcagtgtc atcgacagat gcagcagcag gcggctactg cacaagctgc
     2041 agcagctgcc caggcagcag ccgtggcagg aaacatccct ggcccaggat cagtaggtgg
     2101 aatagctcca gctatcagtc tgtcagctgc tgctggaatt ggtgttgatg accttcgtcg
     2161 cttatgcata ctcaggatga gttttgtgaa aggctgggga ccggattacc caagacagag
     2221 catcaaagaa acaccttgct ggattgaaat tcacttacac cgggccctcc agctcctaga
     2281 cgaagtactt cataccatgc cgattgcaga cccacaacct ttagactgat tgccaacttt
     2341 cttgtacaaa gttggcatta taagaaagca ttgcttatca atttgttgca acgaacaggt
     2401 cactatcagt caaaataaaa tcattatttg ccatccagct gatatcccct atagtgagtc
     2461 gtattacatg gtcatagctg tttcctggca gctctggccc gtgtctcaaa atctctgatg
     2521 ttacattgca caagataaaa atatatcatc atgcctcctc tagaccagcc aggacagaaa
     2581 tgcctcgact tcgctgctac ccaaggttgc cgggtgacgc acaccgtgga aacggatgaa
     2641 ggcacgaacc cagtggacat aagcctgttc ggttcgtaag ctgtaatgca agtagcgtat
     2701 gcgctcacgc aactggtcca gaaccttgac cgaacgcagc ggtggtaacg gcgcagtggc
     2761 ggttttcatg gcttgttatg actgtttttt tggggtacag tctatgcctc gggcatccaa
     2821 gcagcaagcg cgttacgccg tgggtcgatg tttgatgtta tggagcagca acgatgttac
     2881 gcagcagggc agtcgcccta aaacaaagtt aaacattatg agggaagcgg tgatcgccga
     2941 agtatcgact caactatcag aggtagttgg cgtcatcgag cgccatctcg aaccgacgtt
     3001 gctggccgta catttgtacg gctccgcagt ggatggcggc ctgaagccac acagtgatat
     3061 tgatttgctg gttacggtga ccgtaaggct tgatgaaaca acgcggcgag ctttgatcaa
     3121 cgaccttttg gaaacttcgg cttcccctgg agagagcgag attctccgcg ctgtagaagt
     3181 caccattgtt gtgcacgacg acatcattcc gtggcgttat ccagctaagc gcgaactgca
     3241 atttggagaa tggcagcgca atgacattct tgcaggtatc ttcgagccag ccacgatcga
     3301 cattgatctg gctatcttgc tgacaaaagc aagagaacat agcgttgcct tggtaggtcc
     3361 agcggcggag gaactctttg atccggttcc tgaacaggat ctatttgagg cgctaaatga
     3421 aaccttaacg ctatggaact cgccgcccga ctgggctggc gatgagcgaa atgtagtgct
     3481 tacgttgtcc cgcatttggt acagcgcagt aaccggcaaa atcgcgccga aggatgtcgc
     3541 tgccgactgg gcaatggagc gcctgccggc ccagtatcag cccgtcatac ttgaagctag
     3601 acaggcttat cttggacaag aagaagatcg cttggcctcg cgcgcagatc agttggaaga
     3661 atttgtccac tacgtgaaag gcgagatcac caaggtagtc ggcaaataac cctcgagcca
     3721 cccatgacca aaatccctta acgtgagtta cgcgtcgttc cactgagcgt cagaccccgt
     3781 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     3841 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     3901 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     3961 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     4021 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     4081 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     4141 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     4201 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     4261 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     4321 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     4381 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     4441 tgctcacatg tt
//
