LOCUS       sequence-168732-        5520 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway Donor vector containing GEN1 , part of the Target 
            Accelerator Plasmid Collection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5520)
  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 5520)
  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_GEN1_WT
FEATURES             Location/Qualifiers
     source          1..5520
                     /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(2920..3019)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3036..3055)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3037..3055)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3060..3076)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3060..3076)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3507..4298
                     /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      4391..4979
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4880..4899
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5133..5150
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(5309..5336)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(5428..5514)
                     /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 catgggagtg aatgacttgt ggcaaatttt ggagcctgtt aagcaacaca
      241 tccccttgcg taatcttggt gggaaaacca ttgcagttga tctgagtctc tgggtgtgtg
      301 aggcacagac agtcaaaaaa atgatgggca gcgtcatgaa gccccacctc aggaacttat
      361 tttttcgtat ctcatattta acacaaatgg atgtaaaact ggtatttgtt atggaagggg
      421 aaccaccaaa gctgaaagct gatgtcataa gcaagaggaa tcagactcgg tatgggtctt
      481 ctggaaaatc gtggtctcag aaaacaggga gatcacattt taaatcagtc ttaagagagt
      541 gcctccatat gctcgaatgc ttaggaatcc cctgggttca ggctgctggg gaagctgaag
      601 ccatgtgtgc ttatctcaat gctggtggtc atgtcgatgg ctgcctcacc aatgatggag
      661 atactttcct ttatggggcc cagactgttt acaggaattt cactatgaat acaaaggacc
      721 cacatgttga ctgttacaca atgtcatcta tcaagagtaa actaggtttg gatagagatg
      781 ctctggttgg attagcaata cttcttggct gtgattatct cccaaaggga gtccctggag
      841 ttggaaaaga gcaagcatta aaacttatac agattttgaa agggcaaagt ttacttcaga
      901 ggtttaatcg gtggaatgaa acatcttgta actctagtcc acaactgcta gtcactaaaa
      961 aactggctca ttgttccgta tgttcccatc caggttcacc taaggatcat gaacgtaatg
     1021 gatgcagatt atgtaaaagt gataaatatt gtgagccaca tgactatgaa tactgctgtc
     1081 cttgtgagtg gcaccgtaca gaacatgata ggcaactcaa tgaagtagag aacaatatta
     1141 agaagaaagc ttgctgttgt gagggattcc cattccatga ggttattcaa gaattccttt
     1201 taaacaagga taaattggtg aaggttatca ggtaccaaag acctgatttg ttattgtttc
     1261 agagatttac tcttgaaaaa atggagtggc ccaatcacta tgcatgtgag aaattgctgg
     1321 tacttttgac ccattatgac atgatagaaa gaaagcttgg tagcagaaac tctaatcaac
     1381 tacagccaat tcgaattgtt aagactcgaa tcagaaatgg agttcattgt tttgaaatag
     1441 aatgggaaaa gcctgaacat tatgctatgg aagataaaca acatggagaa tttgctttat
     1501 taacaattga ggaagaatca ttgtttgaag cagcatatcc tgagatcgtt gctgtttacc
     1561 aaaaacaaaa gttagaaatt aaagggaaga aacaaaaacg tattaagcct aaagaaaaca
     1621 atttgccaga accagatgaa gtaatgagct ttcagtcaca catgacttta aaacccacat
     1681 gtgaaatctt tcataagcag aattccaagt taaattcggg gatttcccct gatcctacat
     1741 taccacagga atctatttct gcctcattga atagcttgct tttacctaaa aatactccat
     1801 gtttgaatgc acaagaacag ttcatgtctt ctctaagacc tttggctata cagcaaatta
     1861 aagctgtcag taagtctcta atttcagaat ctagtcaacc caatacctca tctcataata
     1921 tatccgtgat tgctgatcta cacttgagca ctattgactg ggaaggtact tcttttagta
     1981 attctccagc tattcaaagg aatacttttt ctcatgattt aaaatcagaa gttgaatcag
     2041 agctatcagc catccctgat ggctttgaaa atatcccaga acaactgtcc tgtgaatcag
     2101 aaaggtacac tgcaaacata aagaaagtgt tggatgagga ttctgatggg attagtcctg
     2161 aagagcatct actttctggc attactgatt tatgtcttca ggatttgcct ttaaaggaac
     2221 gaatatttac aaaattatca tatcctcagg ataatctaca accagatgtc aacctgaaaa
     2281 ctttgtccat acttagtgta aaagaatctt gtattgctaa cagtggttct gattgtacat
     2341 cacatctttc aaaggatctt ccaggaattc ccttgcaaaa tgaatccaga gactctaaaa
     2401 ttctaaaagg agaccagctg cttcaagaag actataaagt caatacttct gtcccttatt
     2461 ctgtcagtaa cacagtggta aagacctgca atgttagacc accaaatact gctttagatc
     2521 atagtagaaa agttgatatg caaaccactc ggaaaatttt aatgaagaag agtgtttgcc
     2581 ttgacagaca ttcctctgat gaacaaagtg ccccagtgtt tgggaaagct aagtacacaa
     2641 ctcaaagaat gaagcacagt tctcaaaagc ataattcatc ccatttcaaa gaaagtggcc
     2701 ataacaagtt gagtagccct aagatacata ttaaagaaac tgaacagtgt gtcagatctt
     2761 atgaaacagc tgaaaatgaa gaaagctgtt tcccagattc aacaaaaagt tctctgagtt
     2821 ctctacaatg tcataagaaa gaaaacaact ctggtacttg tttggatagc cctcttcctt
     2881 tacgccagag attaaaacta agattccaaa gcacttgatt gccaactttc ttgtacaaag
     2941 ttggcattat aagaaagcat tgcttatcaa tttgttgcaa cgaacaggtc actatcagtc
     3001 aaaataaaat cattatttgc catccagctg atatccccta tagtgagtcg tattacatgg
     3061 tcatagctgt ttcctggcag ctctggcccg tgtctcaaaa tctctgatgt tacattgcac
     3121 aagataaaaa tatatcatca tgcctcctct agaccagcca ggacagaaat gcctcgactt
     3181 cgctgctacc caaggttgcc gggtgacgca caccgtggaa acggatgaag gcacgaaccc
     3241 agtggacata agcctgttcg gttcgtaagc tgtaatgcaa gtagcgtatg cgctcacgca
     3301 actggtccag aaccttgacc gaacgcagcg gtggtaacgg cgcagtggcg gttttcatgg
     3361 cttgttatga ctgttttttt ggggtacagt ctatgcctcg ggcatccaag cagcaagcgc
     3421 gttacgccgt gggtcgatgt ttgatgttat ggagcagcaa cgatgttacg cagcagggca
     3481 gtcgccctaa aacaaagtta aacattatga gggaagcggt gatcgccgaa gtatcgactc
     3541 aactatcaga ggtagttggc gtcatcgagc gccatctcga accgacgttg ctggccgtac
     3601 atttgtacgg ctccgcagtg gatggcggcc tgaagccaca cagtgatatt gatttgctgg
     3661 ttacggtgac cgtaaggctt gatgaaacaa cgcggcgagc tttgatcaac gaccttttgg
     3721 aaacttcggc ttcccctgga gagagcgaga ttctccgcgc tgtagaagtc accattgttg
     3781 tgcacgacga catcattccg tggcgttatc cagctaagcg cgaactgcaa tttggagaat
     3841 ggcagcgcaa tgacattctt gcaggtatct tcgagccagc cacgatcgac attgatctgg
     3901 ctatcttgct gacaaaagca agagaacata gcgttgcctt ggtaggtcca gcggcggagg
     3961 aactctttga tccggttcct gaacaggatc tatttgaggc gctaaatgaa accttaacgc
     4021 tatggaactc gccgcccgac tgggctggcg atgagcgaaa tgtagtgctt acgttgtccc
     4081 gcatttggta cagcgcagta accggcaaaa tcgcgccgaa ggatgtcgct gccgactggg
     4141 caatggagcg cctgccggcc cagtatcagc ccgtcatact tgaagctaga caggcttatc
     4201 ttggacaaga agaagatcgc ttggcctcgc gcgcagatca gttggaagaa tttgtccact
     4261 acgtgaaagg cgagatcacc aaggtagtcg gcaaataacc ctcgagccac ccatgaccaa
     4321 aatcccttaa cgtgagttac gcgtcgttcc actgagcgtc agaccccgta gaaaagatca
     4381 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     4441 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     4501 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     4561 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     4621 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     4681 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     4741 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     4801 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     4861 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     4921 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     4981 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     5041 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     5101 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     5161 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
     5221 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat acgcgtaccg
     5281 ctagccagga agagtttgta gaaacgcaaa aaggccatcc gtcaggatgg ccttctgctt
     5341 agtttgatgc ctggcagttt atggcgggcg tcctgcccgc caccctccgg gccgttgctt
     5401 cacaacgttc aaatccgctc ccggcggatt tgtcctactc aggagagcgt tcaccgacaa
     5461 acaacagata aaacgaaagg cccagtcttc cgactgagcc tttcgtttta tttgatgcct
//
