LOCUS       sequence-166518-        3435 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway Donor vector containing VHL, part of the Target Accelerator 
            Plasmid Collection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3435)
  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 3435)
  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_VHL_p.E186K
FEATURES             Location/Qualifiers
     source          1..3435
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      40..628
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     529..548
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     782..799
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(958..985)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(1077..1163)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     1212..1234
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1226..1243
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1227..1243
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1259..1358
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(2004..2103)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2120..2139)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2121..2139)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2144..2160)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2144..2160)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2591..3382
                     /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"
ORIGIN
        1 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
       61 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
      121 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
      181 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
      241 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
      301 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
      361 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
      421 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
      481 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
      541 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
      601 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
      661 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
      721 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
      781 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
      841 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
      901 tgagcgcaac gcaattaata cgcgtaccgc tagccaggaa gagtttgtag aaacgcaaaa
      961 aggccatccg tcaggatggc cttctgctta gtttgatgcc tggcagttta tggcgggcgt
     1021 cctgcccgcc accctccggg ccgttgcttc acaacgttca aatccgctcc cggcggattt
     1081 gtcctactca ggagagcgtt caccgacaaa caacagataa aacgaaaggc ccagtcttcc
     1141 gactgagcct ttcgttttat ttgatgcctg gcagttccct actctcgcgt taacgctagc
     1201 atggatgttt tcccagtcac gacgttgtaa aacgacggcc agtcttaagc tcgggcccca
     1261 aataatgatt ttattttgac tgatagtgac ctgttcgttg caacaaattg atgagcaatg
     1321 cttttttata atgccaactt tgtacaaaaa agttggcacc atgccccgga gggcggagaa
     1381 ctgggacgag gccgaggtag gcgcggagga ggcaggcgtc gaagagtacg gccctgaaga
     1441 agacggcggg gaggagtcgg gcgccgagga gtccggcccg gaagagtccg gcccggagga
     1501 actgggcgcc gaggaggaga tggaggccgg gcggccgcgg cccgtgctgc gctcggtgaa
     1561 ctcgcgcgag ccctcccagg tcatcttctg caatcgcagt ccgcgcgtcg tgctgcccgt
     1621 atggctcaac ttcgacggcg agccgcagcc ctacccaacg ctgccgcctg gcacgggccg
     1681 ccgcatccac agctaccgag gtcacctttg gctcttcaga gatgcaggga cacacgatgg
     1741 gcttctggtt aaccaaactg aattatttgt gccatctctc aatgttgacg gacagcctat
     1801 ttttgccaat atcacactgc cagtgtatac tctgaaagag cgatgcctcc aggttgtccg
     1861 gagcctagtc aagcctgaga attacaggag actggacatc gtcaggtcgc tctacaaaga
     1921 tctggaagac cacccaaatg tgcagaaaga cctggagcgg ctgacacagg agcgcattgc
     1981 acatcaacgg atgggagatt gattgccaac tttcttgtac aaagttgtca ttataagaaa
     2041 gcattgctta tcaatttgtt gcaacgaaca ggtcactatc agtcaaaata aaatcattat
     2101 ttgccatcca gctgatatcc cctatagtga gtcgtattac atggtcatag ctgtttcctg
     2161 gcagctctgg cccgtgtctc aaaatctctg atgttacatt gcacaagata aaaatatatc
     2221 atcatgcctc ctctagacca gccaggacag aaatgcctcg acttcgctgc tacccaaggt
     2281 tgccgggtga cgcacaccgt ggaaacggat gaaggcacga acccagtgga cataagcctg
     2341 ttcggttcgt aagctgtaat gcaagtagcg tatgcgctca cgcaactggt ccagaacctt
     2401 gaccgaacgc agcggtggta acggcgcagt ggcggttttc atggcttgtt atgactgttt
     2461 ttttggggta cagtctatgc ctcgggcatc caagcagcaa gcgcgttacg ccgtgggtcg
     2521 atgtttgatg ttatggagca gcaacgatgt tacgcagcag ggcagtcgcc ctaaaacaaa
     2581 gttaaacatt atgagggaag cggtgatcgc cgaagtatcg actcaactat cagaggtagt
     2641 tggcgtcatc gagcgccatc tcgaaccgac gttgctggcc gtacatttgt acggctccgc
     2701 agtggatggc ggcctgaagc cacacagtga tattgatttg ctggttacgg tgaccgtaag
     2761 gcttgatgaa acaacgcggc gagctttgat caacgacctt ttggaaactt cggcttcccc
     2821 tggagagagc gagattctcc gcgctgtaga agtcaccatt gttgtgcacg acgacatcat
     2881 tccgtggcgt tatccagcta agcgcgaact gcaatttgga gaatggcagc gcaatgacat
     2941 tcttgcaggt atcttcgagc cagccacgat cgacattgat ctggctatct tgctgacaaa
     3001 agcaagagaa catagcgttg ccttggtagg tccagcggcg gaggaactct ttgatccggt
     3061 tcctgaacag gatctatttg aggcgctaaa tgaaacctta acgctatgga actcgccgcc
     3121 cgactgggct ggcgatgagc gaaatgtagt gcttacgttg tcccgcattt ggtacagcgc
     3181 agtaaccggc aaaatcgcgc cgaaggatgt cgctgccgac tgggcaatgg agcgcctgcc
     3241 ggcccagtat cagcccgtca tacttgaagc tagacaggct tatcttggac aagaagaaga
     3301 tcgcttggcc tcgcgcgcag atcagttgga agaatttgtc cactacgtga aaggcgagat
     3361 caccaaggta gtcggcaaat aaccctcgag ccacccatga ccaaaatccc ttaacgtgag
     3421 ttacgcgtcg ttcca
//
