LOCUS       sequence-168822-        3699 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway Donor vector containing CDK4 , part of the Target 
            Accelerator Plasmid Collection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3699)
  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 3699)
  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_CDK4_WT
FEATURES             Location/Qualifiers
     source          1..3699
                     /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(1578..1677)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1694..1713)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1695..1713)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1718..1734)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1718..1734)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2165..2956
                     /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      3049..3637
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3538..3557
                     /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 gctacctctc gatatgagcc agtggctgaa attggtgtcg gtgcctatgg
      721 gacagtgtac aaggcccgtg atccccacag tggccacttt gtggccctca agagtgtgag
      781 agtccccaat ggaggaggag gtggaggagg ccttcccatc agcacagttc gtgaggtggc
      841 tttactgagg cgactggagg cttttgagca tcccaatgtt gtccggctga tggacgtctg
      901 tgccacatcc cgaactgacc gggagatcaa ggtaaccctg gtgtttgagc atgtagacca
      961 ggacctaagg acatatctgg acaaggcacc cccaccaggc ttgccagccg aaacgatcaa
     1021 ggatctgatg cgccagtttc taagaggcct agatttcctt catgccaatt gcatcgttca
     1081 ccgagatctg aagccagaga acattctggt gacaagtggt ggaacagtca agctggctga
     1141 ctttggcctg gccagaatct acagctacca gatggcactt acacccgtgg ttgttacact
     1201 ctggtaccga gctcccgaag ttcttctgca gtccacatat gcaacacctg tggacatgtg
     1261 gagtgttggc tgtatctttg cagagatgtt tcgtcgaaag cctctcttct gtggaaactc
     1321 tgaagccgac cagttgggca aaatctttga cctgattggg ctgcctccag aggatgactg
     1381 gcctcgagat gtatccctgc cccgtggagc ctttcccccc agagggcccc gcccagtgca
     1441 gtcggtggta cctgagatgg aggagtcggg agcacagctg ctgctggaaa tgctgacttt
     1501 taacccacac aagcgaatct ctgcctttcg agctctgcag cactcttatc tacataagga
     1561 tgaaggtaat ccggagtgcc caactttctt gtacaaagtt ggcattataa gaaagcattg
     1621 cttatcaatt tgttgcaacg aacaggtcac tatcagtcaa aataaaatca ttatttgcca
     1681 tccagctgat atcccctata gtgagtcgta ttacatggtc atagctgttt cctggcagct
     1741 ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa gataaaaata tatcatcatg
     1801 cctcctctag accagccagg acagaaatgc ctcgacttcg ctgctaccca aggttgccgg
     1861 gtgacgcaca ccgtggaaac ggatgaaggc acgaacccag tggacataag cctgttcggt
     1921 tcgtaagctg taatgcaagt agcgtatgcg ctcacgcaac tggtccagaa ccttgaccga
     1981 acgcagcggt ggtaacggcg cagtggcggt tttcatggct tgttatgact gtttttttgg
     2041 ggtacagtct atgcctcggg catccaagca gcaagcgcgt tacgccgtgg gtcgatgttt
     2101 gatgttatgg agcagcaacg atgttacgca gcagggcagt cgccctaaaa caaagttaaa
     2161 cattatgagg gaagcggtga tcgccgaagt atcgactcaa ctatcagagg tagttggcgt
     2221 catcgagcgc catctcgaac cgacgttgct ggccgtacat ttgtacggct ccgcagtgga
     2281 tggcggcctg aagccacaca gtgatattga tttgctggtt acggtgaccg taaggcttga
     2341 tgaaacaacg cggcgagctt tgatcaacga ccttttggaa acttcggctt cccctggaga
     2401 gagcgagatt ctccgcgctg tagaagtcac cattgttgtg cacgacgaca tcattccgtg
     2461 gcgttatcca gctaagcgcg aactgcaatt tggagaatgg cagcgcaatg acattcttgc
     2521 aggtatcttc gagccagcca cgatcgacat tgatctggct atcttgctga caaaagcaag
     2581 agaacatagc gttgccttgg taggtccagc ggcggaggaa ctctttgatc cggttcctga
     2641 acaggatcta tttgaggcgc taaatgaaac cttaacgcta tggaactcgc cgcccgactg
     2701 ggctggcgat gagcgaaatg tagtgcttac gttgtcccgc atttggtaca gcgcagtaac
     2761 cggcaaaatc gcgccgaagg atgtcgctgc cgactgggca atggagcgcc tgccggccca
     2821 gtatcagccc gtcatacttg aagctagaca ggcttatctt ggacaagaag aagatcgctt
     2881 ggcctcgcgc gcagatcagt tggaagaatt tgtccactac gtgaaaggcg agatcaccaa
     2941 ggtagtcggc aaataaccct cgagccaccc atgaccaaaa tcccttaacg tgagttacgc
     3001 gtcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     3061 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     3121 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     3181 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     3241 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     3301 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     3361 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     3421 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     3481 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     3541 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     3601 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     3661 acggttcctg gccttttgct ggccttttgc tcacatgtt
//
