LOCUS       sequence-165301-        5655 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway Donor vector containing CYLD , part of the Target 
            Accelerator Plasmid Collection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5655)
  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 5655)
  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_CYLD_WT
FEATURES             Location/Qualifiers
     source          1..5655
                     /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(3055..3154)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3171..3190)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3172..3190)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3195..3211)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3195..3211)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3642..4433
                     /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      4526..5114
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5015..5034
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5268..5285
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(5444..5471)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(5563..5649)
                     /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 catgagttca ggcttatgga gccaagaaaa agtcacttca ccctactggg
      241 aagagcggat tttttacttg cttcttcaag aatgcagcgt tacagacaaa caaacacaaa
      301 agctccttaa agtaccgaag ggaagtatag gacagtatat tcaagatcgt tctgtggggc
      361 attcaaggat tccttctgca aaaggcaaga aaaatcagat tggattaaaa attctagagc
      421 aacctcatgc agttctcttt gttgatgaaa aggatgttgt agagataaat gaaaagttca
      481 cagagttact tttggcaatt accaattgtg aggagaggtt cagcctgttt aaaaacagaa
      541 acagactaag taaaggcctc caaatagacg tgggctgtcc tgtgaaagta cagctgagat
      601 ctggggaaga aaaatttcct ggagttgtac gcttcagagg acccctgtta gcagagagga
      661 cagtctccgg aatattcttt ggagttgaat tgctggaaga aggtcgtggt caaggtttca
      721 ctgacggggt gtaccaaggg aaacagcttt ttcagtgtga tgaagattgt ggcgtgtttg
      781 ttgcattgga caagctagaa ctcatagaag atgatgacac tgcattggaa agtgattacg
      841 caggtcctgg ggacacaatg caggtcgaac ttcctccttt ggaaataaac tccagagttt
      901 ctttgaaggt tggagaaaca atagaatctg gaacagttat attctgtgat gttttgccag
      961 gaaaagaaag cttaggatat tttgttggtg tggacatgga taaccctatt ggcaactggg
     1021 atggaagatt tgatggagtg cagctttgta gttttgcgtg tgttgaaagt acaattctat
     1081 tgcacatcaa tgatatcatc ccagagagtg tgacgcagga aaggaggcct cccaaacttg
     1141 cctttatgtc aagaggtgtt ggggacaaag gttcatccag tcataataaa ccaaaggcta
     1201 caggatctac ctcagaccct ggaaatagaa acagatctga attattttat accttaaatg
     1261 ggtcttctgt tgactcacaa ccacaatcca aatcaaaaaa tacatggtac attgatgaag
     1321 ttgcagaaga ccctgcaaaa tctcttacag agatatctac agactttgac cgttcttcac
     1381 caccactcca gcctcctcct gtgaactcac tgaccaccga gaacagattc cactctttac
     1441 cattcagtct caccaagatg cccaatacca atggaagtat tggccacagt ccactttctc
     1501 tgtcagccca gtctgtaatg gaagagctaa acactgcacc cgtccaagag agtccaccct
     1561 tggccatgcc tcctgggaac tcacatggtc tagaagtggg ctcattggct gaagttaagg
     1621 agaaccctcc tttctatggg gtaatccgtt ggatcggtca gccaccagga ctgaatgaag
     1681 tgctcgctgg actggaactg gaagatgagt gtgcaggctg tacggatgga accttcagag
     1741 gcactcggta tttcacctgt gccctgaaga aggcgctgtt tgtgaaactg aagagctgca
     1801 ggcctgactc taggtttgca tcattgcagc cggtttccaa tcagattgag cgctgtaact
     1861 ctttagcatt tggaggctac ttaagtgaag tagtagaaga aaatactcca ccaaaaatgg
     1921 aaaaagaagg cttggagata atgattggga agaagaaagg catccagggt cattacaatt
     1981 cttgttactt agactcaacc ttattctgct tatttgcttt tagttctgtt ctggacactg
     2041 tgttacttag acccaaagaa aagaacgatg tagaatatta tagtgaaacc caagagctac
     2101 tgaggacaga aattgttaat cctctgagaa tatatggata tgtgtgtgcc acaaaaatta
     2161 tgaaactgag gaaaatactt gaaaaggtgg aggctgcatc aggatttacc tctgaagaaa
     2221 aagatcctga ggaattcttg aatattctgt ttcatcatat tttaagggta gaacctttgc
     2281 taaaaataag atcagcaggt caaaaggtac aagattgtta cttctatcaa atttttatgg
     2341 aaaaaaatga gaaagttggc gttcccacaa ttcagcagtt gttagaatgg tcttttatca
     2401 acagtaacct gaaatttgca gaggcaccat catgtctgat tattcagatg cctcgatttg
     2461 gaaaagactt taaactattt aaaaaaattt ttccttctct ggaattaaat ataacagatt
     2521 tacttgaaga cactcccaga cagtgccgga tatgtggagg gcttgcaatg tatgagtgta
     2581 gagaatgcta cgacgatccg gacatctcag ctggaaaaat caagcagttt tgtaaaacct
     2641 gcaacactca agtccacctt catccgaaga ggctgaatca taaatataac ccagtgtcac
     2701 ttcccaaaga cttacccgac tgggactgga gacacggctg catcccttgc cagaatatgg
     2761 agttatttgc tgttctctgc atagaaacaa gccactatgt tgcttttgtg aagtatggga
     2821 aggacgattc tgcctggctc ttctttgaca gcatggccga tcgggatggt ggtcagaatg
     2881 gcttcaacat tcctcaagtc accccatgcc cagaagtagg agagtacttg aagatgtctc
     2941 tggaagacct gcattccttg gactccagga gaatccaagg ctgtgcacga agactgcttt
     3001 gtgatgcata tatgtgcatg taccagagtc caacaatgag tttgtacaaa taattgccaa
     3061 ctttcttgta caaagttggc attataagaa agcattgctt atcaatttgt tgcaacgaac
     3121 aggtcactat cagtcaaaat aaaatcatta tttgccatcc agctgatatc ccctatagtg
     3181 agtcgtatta catggtcata gctgtttcct ggcagctctg gcccgtgtct caaaatctct
     3241 gatgttacat tgcacaagat aaaaatatat catcatgcct cctctagacc agccaggaca
     3301 gaaatgcctc gacttcgctg ctacccaagg ttgccgggtg acgcacaccg tggaaacgga
     3361 tgaaggcacg aacccagtgg acataagcct gttcggttcg taagctgtaa tgcaagtagc
     3421 gtatgcgctc acgcaactgg tccagaacct tgaccgaacg cagcggtggt aacggcgcag
     3481 tggcggtttt catggcttgt tatgactgtt tttttggggt acagtctatg cctcgggcat
     3541 ccaagcagca agcgcgttac gccgtgggtc gatgtttgat gttatggagc agcaacgatg
     3601 ttacgcagca gggcagtcgc cctaaaacaa agttaaacat tatgagggaa gcggtgatcg
     3661 ccgaagtatc gactcaacta tcagaggtag ttggcgtcat cgagcgccat ctcgaaccga
     3721 cgttgctggc cgtacatttg tacggctccg cagtggatgg cggcctgaag ccacacagtg
     3781 atattgattt gctggttacg gtgaccgtaa ggcttgatga aacaacgcgg cgagctttga
     3841 tcaacgacct tttggaaact tcggcttccc ctggagagag cgagattctc cgcgctgtag
     3901 aagtcaccat tgttgtgcac gacgacatca ttccgtggcg ttatccagct aagcgcgaac
     3961 tgcaatttgg agaatggcag cgcaatgaca ttcttgcagg tatcttcgag ccagccacga
     4021 tcgacattga tctggctatc ttgctgacaa aagcaagaga acatagcgtt gccttggtag
     4081 gtccagcggc ggaggaactc tttgatccgg ttcctgaaca ggatctattt gaggcgctaa
     4141 atgaaacctt aacgctatgg aactcgccgc ccgactgggc tggcgatgag cgaaatgtag
     4201 tgcttacgtt gtcccgcatt tggtacagcg cagtaaccgg caaaatcgcg ccgaaggatg
     4261 tcgctgccga ctgggcaatg gagcgcctgc cggcccagta tcagcccgtc atacttgaag
     4321 ctagacaggc ttatcttgga caagaagaag atcgcttggc ctcgcgcgca gatcagttgg
     4381 aagaatttgt ccactacgtg aaaggcgaga tcaccaaggt agtcggcaaa taaccctcga
     4441 gccacccatg accaaaatcc cttaacgtga gttacgcgtc gttccactga gcgtcagacc
     4501 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     4561 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     4621 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     4681 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     4741 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     4801 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     4861 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     4921 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     4981 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     5041 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     5101 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     5161 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     5221 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     5281 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     5341 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
     5401 ttaatacgcg taccgctagc caggaagagt ttgtagaaac gcaaaaaggc catccgtcag
     5461 gatggccttc tgcttagttt gatgcctggc agtttatggc gggcgtcctg cccgccaccc
     5521 tccgggccgt tgcttcacaa cgttcaaatc cgctcccggc ggatttgtcc tactcaggag
     5581 agcgttcacc gacaaacaac agataaaacg aaaggcccag tcttccgact gagcctttcg
     5641 ttttatttga tgcct
//
