LOCUS       sequence-168756-        4578 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway Donor vector containing EYS ,part of the Target Accelerator 
            Plasmid Collection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4578)
  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 4578)
  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_EYS_WT
FEATURES             Location/Qualifiers
     source          1..4578
                     /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(1978..2077)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2094..2113)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2095..2113)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2118..2134)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2118..2134)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2565..3356
                     /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      3449..4037
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3938..3957
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4191..4208
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4367..4394)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4486..4572)
                     /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 aagttggcac catgactgac aaatcaatcg tcattctgag cctgatggtt tttcacagct
      241 ctttcataaa tggaaaaaca tgtagacggc aattggtgga agaatggcat ccacaaccct
      301 catcatatgt ggtaaattgg acactaacag aaaacatctg cttggacttc tacagagatt
      361 gctggttttt gggtgtaaac actaaaatag atacttcagg caatcaagct gttccccaga
      421 tttgcccttt gcaaattcaa ttaggagata tccttgtaat ttcttctgaa ccatctcttc
      481 aatttccaga aataaatttg atgaatgttt ctgaaacatc tttcgttggc tgtgtgcaaa
      541 ataccacaac ggaagatcag ttactttttg gctgcagact aaaaggaatg cacactgtta
      601 attctaagtg gctgagtgtt gggacacatt attttatcac agttatggca agtggtccat
      661 caccttgtcc actgggactt cgactaaatg tgacagtgaa acagcagttc tgccaggaat
      721 ctctgagttc agaattttgc tctggtcatg gtaaatgtct tagtgaagct tggagcaaga
      781 catatagctg ccattgccag cctccatttt ctggaaaata ctgccaggaa cttgatgcat
      841 gttcttttaa accatgtaaa aataatggca gttgcattaa taaaagagaa aattgggatg
      901 agcaagcata tgaatgtgtc tgtcacccac catttacagg aaagaattgc tcagaaataa
      961 ttggccagtg tcaaccacat gtctgtttcc atggaaactg cagcaatatt acttcaaata
     1021 gtttcatttg tgaatgtgat gagcaatttt caggtccatt ctgtgaggtg tcagcaaaac
     1081 cttgtgtttc tctgcttttt tggaaaagag gaatttgccc aaatagcagt tctgcttata
     1141 cttatgaatg cccaaaagga tcttccagcc aaaatggtga aactgatgtc agtgagtttt
     1201 cattagtacc atgtcagaat ggtactgact gcatcaaaat atcaaatgat gttatgtgca
     1261 tctgttcacc aatatttaca gatttgcttt gtaagagcat tcaaacatca tgtgagtcat
     1321 ttcctttgag gaataatgct acatgtaaga aatgtgagaa agattatcct tgcagctgta
     1381 tatcaggatt tactgaaaaa aactgtgaga aagcaattga ccactgtaaa ctgctcagca
     1441 tcaactgtct gaatgaagaa tggtgtttca atataattgg aagattcaaa tatgtatgca
     1501 ttccagggtg cacaaaaaat ccatgttggt ttttgaagaa tgtttaccta attcatcaac
     1561 acctctgcta ctgtggagtc accttccatg gtatttgcca agataaaggt cctgctcaat
     1621 ttgaatatgt gtggcaattg ggatttgcag gatctgaagg cgaaaagtgc caaggggtta
     1681 ttgatgccta tttctttctg gctgcaaact gcactgaaga tgcaacctat gtgaacgatc
     1741 ctgaagataa taattcttca tgttggttcc cacatgaagg cacaaaagag atttgtgcaa
     1801 atggatgcag ttgtttgagt gaagaagaca gtcaggaata tcggtatcta tgttttctca
     1861 gatgggctgg caacatgtat ctggaaaata caactgatga tcaagaaaat gagtgtcaac
     1921 atgaagctgt ttgtaaagat gaaattaata gacccaggta tttatgtatc atttgattgc
     1981 caactttctt gtacaaagtt ggcattataa gaaagcattg cttatcaatt tgttgcaacg
     2041 aacaggtcac tatcagtcaa aataaaatca ttatttgcca tccagctgat atcccctata
     2101 gtgagtcgta ttacatggtc atagctgttt cctggcagct ctggcccgtg tctcaaaatc
     2161 tctgatgtta cattgcacaa gataaaaata tatcatcatg cctcctctag accagccagg
     2221 acagaaatgc ctcgacttcg ctgctaccca aggttgccgg gtgacgcaca ccgtggaaac
     2281 ggatgaaggc acgaacccag tggacataag cctgttcggt tcgtaagctg taatgcaagt
     2341 agcgtatgcg ctcacgcaac tggtccagaa ccttgaccga acgcagcggt ggtaacggcg
     2401 cagtggcggt tttcatggct tgttatgact gtttttttgg ggtacagtct atgcctcggg
     2461 catccaagca gcaagcgcgt tacgccgtgg gtcgatgttt gatgttatgg agcagcaacg
     2521 atgttacgca gcagggcagt cgccctaaaa caaagttaaa cattatgagg gaagcggtga
     2581 tcgccgaagt atcgactcaa ctatcagagg tagttggcgt catcgagcgc catctcgaac
     2641 cgacgttgct ggccgtacat ttgtacggct ccgcagtgga tggcggcctg aagccacaca
     2701 gtgatattga tttgctggtt acggtgaccg taaggcttga tgaaacaacg cggcgagctt
     2761 tgatcaacga ccttttggaa acttcggctt cccctggaga gagcgagatt ctccgcgctg
     2821 tagaagtcac cattgttgtg cacgacgaca tcattccgtg gcgttatcca gctaagcgcg
     2881 aactgcaatt tggagaatgg cagcgcaatg acattcttgc aggtatcttc gagccagcca
     2941 cgatcgacat tgatctggct atcttgctga caaaagcaag agaacatagc gttgccttgg
     3001 taggtccagc ggcggaggaa ctctttgatc cggttcctga acaggatcta tttgaggcgc
     3061 taaatgaaac cttaacgcta tggaactcgc cgcccgactg ggctggcgat gagcgaaatg
     3121 tagtgcttac gttgtcccgc atttggtaca gcgcagtaac cggcaaaatc gcgccgaagg
     3181 atgtcgctgc cgactgggca atggagcgcc tgccggccca gtatcagccc gtcatacttg
     3241 aagctagaca ggcttatctt ggacaagaag aagatcgctt ggcctcgcgc gcagatcagt
     3301 tggaagaatt tgtccactac gtgaaaggcg agatcaccaa ggtagtcggc aaataaccct
     3361 cgagccaccc atgaccaaaa tcccttaacg tgagttacgc gtcgttccac tgagcgtcag
     3421 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     3481 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     3541 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     3601 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     3661 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     3721 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     3781 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     3841 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     3901 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     3961 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     4021 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     4081 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     4141 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     4201 tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
     4261 ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg
     4321 caattaatac gcgtaccgct agccaggaag agtttgtaga aacgcaaaaa ggccatccgt
     4381 caggatggcc ttctgcttag tttgatgcct ggcagtttat ggcgggcgtc ctgcccgcca
     4441 ccctccgggc cgttgcttca caacgttcaa atccgctccc ggcggatttg tcctactcag
     4501 gagagcgttc accgacaaac aacagataaa acgaaaggcc cagtcttccg actgagcctt
     4561 tcgttttatt tgatgcct
//
