LOCUS       sequence-164543-        4609 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway Donor vector containing ARAF , part of the Target 
            Accelerator Plasmid Collection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4609)
  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 4609)
  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_ARAF_WT
FEATURES             Location/Qualifiers
     source          1..4609
                     /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(2488..2587)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2604..2623)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2605..2623)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2628..2644)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2628..2644)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3075..3866
                     /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      3959..4547
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4448..4467
                     /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 gagccaccac ggggcccccc tgccaatggg gccgagccat cccgggcagt
      721 gggcaccgtc aaagtatacc tgcccaacaa gcaacgcacg gtggtgactg tccgggatgg
      781 catgagtgtc tacgactctc tagacaaggc cctgaaggtg cggggtctaa atcaggactg
      841 ctgtgtggtc taccgactca tcaagggacg aaagacggtc actgcctggg acacagccat
      901 tgctcccctg gatggcgagg agctcattgt cgaggtcctt gaagatgtcc cgctgaccat
      961 gcacaatttt gtacggaaga ccttcttcag cctggcgttc tgtgacttct gccttaagtt
     1021 tctgttccat ggcttccgtt gccaaacctg tggctacaag ttccaccagc attgttcctc
     1081 caaggtcccc acagtctgtg ttgacatgag taccaaccgc caacagttct accacagtgt
     1141 ccaggatttg tccggaggct ccagacagca tgaggctccc tcgaaccgcc ccctgaatga
     1201 gttgctaacc ccccagggtc ccagcccccg cacccagcac tgtgacccgg agcacttccc
     1261 cttccctgcc ccagccaatg cccccctaca gcgcatccgc tccacgtcca ctcccaacgt
     1321 ccatatggtc agcaccacgg cccccatgga ctccaacctc atccagctca ctggccagag
     1381 tttcagcact gatgctgccg gtagtagagg aggtagtgat ggaacccccc gggggagccc
     1441 cagcccagcc agcgtgtcct cggggaggaa gtccccacat tccaagtcac cagcagagca
     1501 gcgcgagcgg aagtccttgg ccgatgacaa gaagaaagtg aagaacctgg ggtaccggga
     1561 ctcaggctat tactgggagg taccacccag tgaggtgcag ctgctgaaga ggatcgggac
     1621 gggctcgttt ggcaccgtgt ttcgagggcg gtggcatggc gatgtggccg tgaaggtgct
     1681 caaggtgtcc cagcccacag ctgagcaggc ccaggctttc aagaatgaga tgcaggtgct
     1741 caggaagacg cgacatgtca acatcttgct gtttatgggc ttcatgaccc ggccgggatt
     1801 tgccatcatc acacagtggt gtgagggctc cagcctctac catcacctgc atgtggccga
     1861 cacacgcttc gacatggtcc agctcatcga cgtggcccgg cagactgccc agggcatgga
     1921 ctacctccat gccaagaaca tcatccaccg agatctcaag tctaacaaca tcttcctaca
     1981 tgaggggctc acggtgaaga tcggtgactt tggcttggcc acagtgaaga ctcgatggag
     2041 cggggcccag cccttggagc agccctcagg atctgtgctg tggatggcag ctgaggtgat
     2101 ccgtatgcag gacccgaacc cctacagctt ccagtcagac gtctatgcct acggggttgt
     2161 gctctacgag cttatgactg gctcactgcc ttacagccac attggctgcc gtgaccagat
     2221 tatctttatg gtgggccgtg gctatctgtc cccggacctc agcaaaatct ccagcaactg
     2281 ccccaaggcc atgcggcgcc tgctgtctga ctgcctcaag ttccagcggg aggagcggcc
     2341 cctcttcccc cagatcctgg ccacaattga gctgctgcaa cggtcactcc ccaagattga
     2401 gcggagtgcc tcggaaccct ccttgcaccg cacccaggcc gatgagttgc ctgcctgcct
     2461 actcagcgca gcccgccttg tgccttagcc caactttctt gtacaaagtt ggcattataa
     2521 gaaagcattg cttatcaatt tgttgcaacg aacaggtcac tatcagtcaa aataaaatca
     2581 ttatttgcca tccagctgat atcccctata gtgagtcgta ttacatggtc atagctgttt
     2641 cctggcagct ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa gataaaaata
     2701 tatcatcatg cctcctctag accagccagg acagaaatgc ctcgacttcg ctgctaccca
     2761 aggttgccgg gtgacgcaca ccgtggaaac ggatgaaggc acgaacccag tggacataag
     2821 cctgttcggt tcgtaagctg taatgcaagt agcgtatgcg ctcacgcaac tggtccagaa
     2881 ccttgaccga acgcagcggt ggtaacggcg cagtggcggt tttcatggct tgttatgact
     2941 gtttttttgg ggtacagtct atgcctcggg catccaagca gcaagcgcgt tacgccgtgg
     3001 gtcgatgttt gatgttatgg agcagcaacg atgttacgca gcagggcagt cgccctaaaa
     3061 caaagttaaa cattatgagg gaagcggtga tcgccgaagt atcgactcaa ctatcagagg
     3121 tagttggcgt catcgagcgc catctcgaac cgacgttgct ggccgtacat ttgtacggct
     3181 ccgcagtgga tggcggcctg aagccacaca gtgatattga tttgctggtt acggtgaccg
     3241 taaggcttga tgaaacaacg cggcgagctt tgatcaacga ccttttggaa acttcggctt
     3301 cccctggaga gagcgagatt ctccgcgctg tagaagtcac cattgttgtg cacgacgaca
     3361 tcattccgtg gcgttatcca gctaagcgcg aactgcaatt tggagaatgg cagcgcaatg
     3421 acattcttgc aggtatcttc gagccagcca cgatcgacat tgatctggct atcttgctga
     3481 caaaagcaag agaacatagc gttgccttgg taggtccagc ggcggaggaa ctctttgatc
     3541 cggttcctga acaggatcta tttgaggcgc taaatgaaac cttaacgcta tggaactcgc
     3601 cgcccgactg ggctggcgat gagcgaaatg tagtgcttac gttgtcccgc atttggtaca
     3661 gcgcagtaac cggcaaaatc gcgccgaagg atgtcgctgc cgactgggca atggagcgcc
     3721 tgccggccca gtatcagccc gtcatacttg aagctagaca ggcttatctt ggacaagaag
     3781 aagatcgctt ggcctcgcgc gcagatcagt tggaagaatt tgtccactac gtgaaaggcg
     3841 agatcaccaa ggtagtcggc aaataaccct cgagccaccc atgaccaaaa tcccttaacg
     3901 tgagttacgc gtcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     3961 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     4021 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     4081 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     4141 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     4201 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     4261 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     4321 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     4381 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     4441 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     4501 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     4561 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgtt
//
