LOCUS       sequence-164661-        4056 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway Donor vector containing GSK3B , part of the Target 
            Accelerator Plasmid Collection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4056)
  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 4056)
  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_GSK3B_WT
FEATURES             Location/Qualifiers
     source          1..4056
                     /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"
     CDS             1488..1499
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     protein_bind    complement(1935..2034)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2051..2070)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2052..2070)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2075..2091)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2075..2091)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2522..3313
                     /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      3406..3994
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3895..3914
                     /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 agttggcacc atgtcagggc ggcccagaac cacctccttt gcggagagct gcaagccggt
      721 gcagcagcct tcagcttttg gcagcatgaa agttagcaga gacaaggacg gcagcaaggt
      781 gacaacagtg gtggcaactc ctgggcaggg tccagacagg ccacaagaag tcagctatac
      841 agacactaaa gtgattggaa atggatcatt tggtgtggta tatcaagcca aactttgtga
      901 ttcaggagaa ctggtcgcca tcaagaaagt attgcaggac aagagattta agaatcgaga
      961 gctccagatc atgagaaagc tagatcactg taacatagtc cgattgcgtt atttcttcta
     1021 ctccagtggt gagaagaaag atgaggtcta tcttaatctg gtgctggact atgttccgga
     1081 aacagtatac agagttgcca gacactatag tcgagccaaa cagacgctcc ctgtgattta
     1141 tgtcaagttg tatatgtatc agctgttccg aagtttagcc tatatccatt cctttggaat
     1201 ctgccatcgg gatattaaac cgcagaacct cttgttggat cctgatactg ctgtattaaa
     1261 actctgtgac tttggaagtg caaagcagct ggtccgagga gaacccaatg tttcgtatat
     1321 ctgttctcgg tactataggg caccagagtt gatctttgga gccactgatt atacctctag
     1381 tatagatgta tggtctgctg gctgtgtgtt ggctgagctg ttactaggac aaccaatatt
     1441 tccaggggat agtggtgtgg atcagttggt agaaataatc aaggtcctgg gaactccaac
     1501 aagggagcaa atcagagaaa tgaacccaaa ctacacagaa tttaaattcc ctcaaattaa
     1561 ggcacatcct tggactaagg tcttccgacc ccgaactcca ccggaggcaa ttgcactgtg
     1621 tagccgtctg ctggagtata caccaactgc ccgactaaca ccactggaag cttgtgcaca
     1681 ttcatttttt gatgaattac gggacccaaa tgtcaaacta ccaaatgggc gagacacacc
     1741 tgcactcttc aacttcacca ctcaagaact gtcaagtaat ccacctctgg ctaccatcct
     1801 tattcctcct catgctcgga ttcaagcagc tgcttcaacc cccacaaatg ccacagcagc
     1861 gtcagatgct aatactggag accgtggaca gaccaataat gctgcttctg catcagcttc
     1921 caactccacc tgcttgccaa ctttcttgta caaagttggc attataagaa agcattgctt
     1981 atcaatttgt tgcaacgaac aggtcactat cagtcaaaat aaaatcatta tttgccatcc
     2041 agctgatatc ccctatagtg agtcgtatta catggtcata gctgtttcct ggcagctctg
     2101 gcccgtgtct caaaatctct gatgttacat tgcacaagat aaaaatatat catcatgcct
     2161 cctctagacc agccaggaca gaaatgcctc gacttcgctg ctacccaagg ttgccgggtg
     2221 acgcacaccg tggaaacgga tgaaggcacg aacccagtgg acataagcct gttcggttcg
     2281 taagctgtaa tgcaagtagc gtatgcgctc acgcaactgg tccagaacct tgaccgaacg
     2341 cagcggtggt aacggcgcag tggcggtttt catggcttgt tatgactgtt tttttggggt
     2401 acagtctatg cctcgggcat ccaagcagca agcgcgttac gccgtgggtc gatgtttgat
     2461 gttatggagc agcaacgatg ttacgcagca gggcagtcgc cctaaaacaa agttaaacat
     2521 tatgagggaa gcggtgatcg ccgaagtatc gactcaacta tcagaggtag ttggcgtcat
     2581 cgagcgccat ctcgaaccga cgttgctggc cgtacatttg tacggctccg cagtggatgg
     2641 cggcctgaag ccacacagtg atattgattt gctggttacg gtgaccgtaa ggcttgatga
     2701 aacaacgcgg cgagctttga tcaacgacct tttggaaact tcggcttccc ctggagagag
     2761 cgagattctc cgcgctgtag aagtcaccat tgttgtgcac gacgacatca ttccgtggcg
     2821 ttatccagct aagcgcgaac tgcaatttgg agaatggcag cgcaatgaca ttcttgcagg
     2881 tatcttcgag ccagccacga tcgacattga tctggctatc ttgctgacaa aagcaagaga
     2941 acatagcgtt gccttggtag gtccagcggc ggaggaactc tttgatccgg ttcctgaaca
     3001 ggatctattt gaggcgctaa atgaaacctt aacgctatgg aactcgccgc ccgactgggc
     3061 tggcgatgag cgaaatgtag tgcttacgtt gtcccgcatt tggtacagcg cagtaaccgg
     3121 caaaatcgcg ccgaaggatg tcgctgccga ctgggcaatg gagcgcctgc cggcccagta
     3181 tcagcccgtc atacttgaag ctagacaggc ttatcttgga caagaagaag atcgcttggc
     3241 ctcgcgcgca gatcagttgg aagaatttgt ccactacgtg aaaggcgaga tcaccaaggt
     3301 agtcggcaaa taaccctcga gccacccatg accaaaatcc cttaacgtga gttacgcgtc
     3361 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     3421 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     3481 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     3541 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     3601 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     3661 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     3721 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     3781 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     3841 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     3901 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     3961 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     4021 gttcctggcc ttttgctggc cttttgctca catgtt
//
