LOCUS       sequence-166776-        4230 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 4230)
  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 4230)
  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_p.V201I
FEATURES             Location/Qualifiers
     source          1..4230
                     /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..2107
                     /codon_start=1
                     /gene="AKT1"
                     /product="human serine/threonine protein kinase activated 
                     by growth factors"
                     /label=Akt1
                     /translation="MSDVAIVKEGWLHKRGEYIKTWRPRYFLLKNDGTFIGYKERPQDV
                     DQREAPLNNFSVAQCQLMKTERPRPNTFIIRCLQWTTVIERTFHVETPEEREEWTTAIQ
                     TVADGLKKQEEEMDFRSGSPSDNSGAEEMEVSLAKPKHRVTMNEFEYLKLLGKGTFGKV
                     ILVKEKATGRYYAMKILKKEVIVAKDEVAHTLTENRILQNSRHPFLTALKYSFQTHDRL
                     CFVMEYANGGELFFHLSRERVFSEDRARFYGAEIVSALDYLHSEKNVVYRDLKLENLML
                     DKDGHIKITDFGLCKEGIKDGATMKTFCGTPEYLAPEVLEDNDYGRAVDWWGLGVVMYE
                     MMCGRLPFYNQDHEKLFELILMEEIRFPRTLGPEAKSLLSGLLKKDPKQRLGGGSEDAK
                     EIMQHRFFAGIVWQHVYEKKLSPPFKPQVTSETDTRYFDEEFTAQMITITPPDQDDSME
                     CVDSERRPHFPQFSYSASGTA"
     protein_bind    complement(2109..2208)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2225..2244)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2226..2244)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2249..2265)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2249..2265)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2696..3487
                     /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      3580..4168
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4069..4088
                     /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 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 aggaggagat
     1021 ggacttccgg tcgggctcac ccagtgacaa ctcaggggct gaagagatgg aggtgtccct
     1081 ggccaagccc aagcaccgcg tgaccatgaa cgagtttgag tacctgaagc tgctgggcaa
     1141 gggcactttc ggcaaggtga tcctggtgaa ggagaaggcc acaggccgct actacgccat
     1201 gaagatcctc aagaaggaag tcatcgtggc caaggacgag gtggcccaca cactcaccga
     1261 gaaccgcatc ctgcagaact ccaggcaccc cttcctcaca gccctgaagt actctttcca
     1321 gacccacgac cgcctctgct ttgtcatgga gtacgccaac gggggcgagc tgttcttcca
     1381 cctgtcccgg gagcgtgtgt tctccgagga ccgggcccgc ttctatggcg ctgagattgt
     1441 gtcagccctg gactacctgc actcggagaa gaacgtggtg taccgggacc tcaagctgga
     1501 gaacctcatg ctggacaagg acgggcacat taagatcaca gacttcgggc tgtgcaagga
     1561 ggggatcaag gacggtgcca ccatgaagac cttttgcggc acacctgagt acctggcccc
     1621 cgaggtgctg gaggacaatg actacggccg tgcagtggac tggtgggggc tgggcgtggt
     1681 catgtacgag atgatgtgcg gtcgcctgcc cttctacaac caggaccatg agaagctttt
     1741 tgagctcatc ctcatggagg agatccgctt cccgcgcacg cttggtcccg aggccaagtc
     1801 cttgctttca gggctgctca agaaggaccc caagcagagg cttggcgggg gctccgagga
     1861 cgccaaggag atcatgcagc atcgcttctt tgccggtatc gtgtggcagc acgtgtacga
     1921 gaagaagctc agcccaccct tcaagcccca ggtcacgtcg gagactgaca ccaggtattt
     1981 tgatgaggag ttcacggccc agatgatcac catcacacca cctgaccaag atgacagcat
     2041 ggagtgtgtg gacagcgagc gcaggcccca cttcccccag ttctcctact cggccagcgg
     2101 cacggcctgc ccaactttct tgtacaaagt tggcattata agaaagcatt gcttatcaat
     2161 ttgttgcaac gaacaggtca ctatcagtca aaataaaatc attatttgcc atccagctga
     2221 tatcccctat agtgagtcgt attacatggt catagctgtt tcctggcagc tctggcccgt
     2281 gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat gcctcctcta
     2341 gaccagccag gacagaaatg cctcgacttc gctgctaccc aaggttgccg ggtgacgcac
     2401 accgtggaaa cggatgaagg cacgaaccca gtggacataa gcctgttcgg ttcgtaagct
     2461 gtaatgcaag tagcgtatgc gctcacgcaa ctggtccaga accttgaccg aacgcagcgg
     2521 tggtaacggc gcagtggcgg ttttcatggc ttgttatgac tgtttttttg gggtacagtc
     2581 tatgcctcgg gcatccaagc agcaagcgcg ttacgccgtg ggtcgatgtt tgatgttatg
     2641 gagcagcaac gatgttacgc agcagggcag tcgccctaaa acaaagttaa acattatgag
     2701 ggaagcggtg atcgccgaag tatcgactca actatcagag gtagttggcg tcatcgagcg
     2761 ccatctcgaa ccgacgttgc tggccgtaca tttgtacggc tccgcagtgg atggcggcct
     2821 gaagccacac agtgatattg atttgctggt tacggtgacc gtaaggcttg atgaaacaac
     2881 gcggcgagct ttgatcaacg accttttgga aacttcggct tcccctggag agagcgagat
     2941 tctccgcgct gtagaagtca ccattgttgt gcacgacgac atcattccgt ggcgttatcc
     3001 agctaagcgc gaactgcaat ttggagaatg gcagcgcaat gacattcttg caggtatctt
     3061 cgagccagcc acgatcgaca ttgatctggc tatcttgctg acaaaagcaa gagaacatag
     3121 cgttgccttg gtaggtccag cggcggagga actctttgat ccggttcctg aacaggatct
     3181 atttgaggcg ctaaatgaaa ccttaacgct atggaactcg ccgcccgact gggctggcga
     3241 tgagcgaaat gtagtgctta cgttgtcccg catttggtac agcgcagtaa ccggcaaaat
     3301 cgcgccgaag gatgtcgctg ccgactgggc aatggagcgc ctgccggccc agtatcagcc
     3361 cgtcatactt gaagctagac aggcttatct tggacaagaa gaagatcgct tggcctcgcg
     3421 cgcagatcag ttggaagaat ttgtccacta cgtgaaaggc gagatcacca aggtagtcgg
     3481 caaataaccc tcgagccacc catgaccaaa atcccttaac gtgagttacg cgtcgttcca
     3541 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     3601 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     3661 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     3721 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     3781 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     3841 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     3901 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     3961 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     4021 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     4081 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     4141 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     4201 ggccttttgc tggccttttg ctcacatgtt
//
