LOCUS       sequence-168876-        4236 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway Donor vector containing AKT1 , part of the Target 
            Accelerator Plasmid Collection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4236)
  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 4236)
  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_AKT1_WT
FEATURES             Location/Qualifiers
     source          1..4236
                     /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             671..2110
                     /codon_start=1
                     /gene="AKT1"
                     /product="human serine/threonine protein kinase activated 
                     by growth factors"
                     /label=Akt1
                     /translation="MSDVAIVKEGWLHKRGEYIKTWRPRYFLLKNDGTFIGYKERPQDV
                     DQREAPLNNFSVAQCQLMKTERPRPNTFIIRCLQWTTVIERTFHVETPEEREEWTTAIQ
                     TVADGLKKQEEEEMDFRSGSPSDNSGAEEMEVSLAKPKHRVTMNEFEYLKLLGKGTFGK
                     VILVKEKATGRYYAMKILKKEVIVAKDEVAHTLTENRVLQNSRHPFLTALKYSFQTHDR
                     LCFVMEYANGGELFFHLSRERVFSEDRARFYGAEIVSALDYLHSEKNVVYRDLKLENLM
                     LDKDGHIKITDFGLCKEGIKDGATMKTFCGTPEYLAPEVLEDNDYGRAVDWWGLGVVMY
                     EMMCGRLPFYNQDHEKLFELILMEEIRFPRTLGPEAKSLLSGLLKKDPKQRLGGGSEDA
                     KEIMQHRFFAGIVWQHVYEKKLSPPFKPQVTSETDTRYFDEEFTAQMITITPPDQDDSM
                     ECVDSERRPHFPQFSYSASGTA"
     protein_bind    complement(2115..2214)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2231..2250)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2232..2250)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2255..2271)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2255..2271)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2702..3493
                     /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      3586..4174
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4075..4094
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 tcgcccaata 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 atgagcgacg tggctattgt gaaggagggt tggctgcaca aacgagggga
      721 gtacatcaag acctggcggc cacgctactt cctcctcaag aatgatggca ccttcattgg
      781 ctacaaggag cggccgcagg atgtggacca acgtgaggct cccctcaaca acttctctgt
      841 ggcgcagtgc cagctgatga agacggagcg gccccggccc aacaccttca tcatccgctg
      901 cctgcagtgg accactgtca tcgaacgcac cttccatgtg gagactcctg aggagcggga
      961 ggagtggaca accgccatcc agactgtggc tgacggcctc aagaagcagg aggaggagga
     1021 gatggacttc cggtcgggct cacccagtga caactcaggg gctgaagaga tggaggtgtc
     1081 cctggccaag cccaagcacc gcgtgaccat gaacgagttt gagtacctga agctgctggg
     1141 caagggcact ttcggcaagg tgatcctggt gaaggagaag gccacaggcc gctactacgc
     1201 catgaagatc ctcaagaagg aagtcatcgt ggccaaggac gaggtggccc acacactcac
     1261 cgagaaccgc gtcctgcaga actccaggca ccccttcctc acagccctga agtactcttt
     1321 ccagacccac gaccgcctct gctttgtcat ggagtacgcc aacgggggcg agctgttctt
     1381 ccacctgtcc cgggagcgtg tgttctccga ggaccgggcc cgcttctatg gcgctgagat
     1441 tgtgtcagcc ctggactacc tgcactcgga gaagaacgtg gtgtaccggg acctcaagct
     1501 ggagaacctc atgctggaca aggacgggca cattaagatc acagacttcg ggctgtgcaa
     1561 ggaggggatc aaggacggtg ccaccatgaa gaccttttgc ggcacacctg agtacctggc
     1621 ccccgaggtg ctggaggaca atgactacgg ccgtgcagtg gactggtggg ggctgggcgt
     1681 ggtcatgtac gagatgatgt gcggtcgcct gcccttctac aaccaggacc atgagaagct
     1741 ttttgagctc atcctcatgg aggagatccg cttcccgcgc acgcttggtc ccgaggccaa
     1801 gtccttgctt tcagggctgc tcaagaagga ccccaagcag aggcttggcg ggggctccga
     1861 ggacgccaag gagatcatgc agcatcgctt ctttgccggt atcgtgtggc agcacgtgta
     1921 cgagaagaag ctcagcccac ccttcaagcc ccaggtcacg tcggagactg acaccaggta
     1981 ttttgatgag gagttcacgg cccagatgat caccatcaca ccacctgacc aagatgacag
     2041 catggagtgt gtggacagcg agcgcaggcc ccacttcccc cagttctcct actcggccag
     2101 cggcacggcc tgattgccaa ctttcttgta caaagttggc attataagaa agcattgctt
     2161 atcaatttgt tgcaacgaac aggtcactat cagtcaaaat aaaatcatta tttgccatcc
     2221 agctgatatc ccctatagtg agtcgtatta catggtcata gctgtttcct ggcagctctg
     2281 gcccgtgtct caaaatctct gatgttacat tgcacaagat aaaaatatat catcatgcct
     2341 cctctagacc agccaggaca gaaatgcctc gacttcgctg ctacccaagg ttgccgggtg
     2401 acgcacaccg tggaaacgga tgaaggcacg aacccagtgg acataagcct gttcggttcg
     2461 taagctgtaa tgcaagtagc gtatgcgctc acgcaactgg tccagaacct tgaccgaacg
     2521 cagcggtggt aacggcgcag tggcggtttt catggcttgt tatgactgtt tttttggggt
     2581 acagtctatg cctcgggcat ccaagcagca agcgcgttac gccgtgggtc gatgtttgat
     2641 gttatggagc agcaacgatg ttacgcagca gggcagtcgc cctaaaacaa agttaaacat
     2701 tatgagggaa gcggtgatcg ccgaagtatc gactcaacta tcagaggtag ttggcgtcat
     2761 cgagcgccat ctcgaaccga cgttgctggc cgtacatttg tacggctccg cagtggatgg
     2821 cggcctgaag ccacacagtg atattgattt gctggttacg gtgaccgtaa ggcttgatga
     2881 aacaacgcgg cgagctttga tcaacgacct tttggaaact tcggcttccc ctggagagag
     2941 cgagattctc cgcgctgtag aagtcaccat tgttgtgcac gacgacatca ttccgtggcg
     3001 ttatccagct aagcgcgaac tgcaatttgg agaatggcag cgcaatgaca ttcttgcagg
     3061 tatcttcgag ccagccacga tcgacattga tctggctatc ttgctgacaa aagcaagaga
     3121 acatagcgtt gccttggtag gtccagcggc ggaggaactc tttgatccgg ttcctgaaca
     3181 ggatctattt gaggcgctaa atgaaacctt aacgctatgg aactcgccgc ccgactgggc
     3241 tggcgatgag cgaaatgtag tgcttacgtt gtcccgcatt tggtacagcg cagtaaccgg
     3301 caaaatcgcg ccgaaggatg tcgctgccga ctgggcaatg gagcgcctgc cggcccagta
     3361 tcagcccgtc atacttgaag ctagacaggc ttatcttgga caagaagaag atcgcttggc
     3421 ctcgcgcgca gatcagttgg aagaatttgt ccactacgtg aaaggcgaga tcaccaaggt
     3481 agtcggcaaa taaccctcga gccacccatg accaaaatcc cttaacgtga gttacgcgtc
     3541 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     3601 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     3661 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     3721 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     3781 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     3841 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     3901 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     3961 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     4021 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     4081 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     4141 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     4201 gttcctggcc ttttgctggc cttttgctca catgtt
//
