LOCUS       sequence-168318-        4233 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 4233)
  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 4233)
  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.Q79K
FEATURES             Location/Qualifiers
     source          1..4233
                     /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
                     DQREAPLNNFSVAQCQLMKTERPRPNTFIIRCLKWTTVIERTFHVETPEEREEWTTAIQ
                     TVADGLKKQEEEEMDFRSGSPSDNSGAEEMEVSLAKPKHRVTMNEFEYLKLLGKGTFGK
                     VILVKEKATGRYYAMKILKKEVIVAKDEVAHTLTENRVLQNSRHPFLTALKYSFQTHDR
                     LCFVMEYANGGELFFHLSRERVFSEDRARFYGAEIVSALDYLHSEKNVVYRDLKLENLM
                     LDKDGHIKITDFGLCKEGIKDGATMKTFCGTPEYLAPEVLEDNDYGRAVDWWGLGVVMY
                     EMMCGRLPFYNQDHEKLFELILMEEIRFPRTLGPEAKSLLSGLLKKDPKQRLGGGSEDA
                     KEIMQHRFFAGIVWQHVYEKKLSPPFKPQVTSETDTRYFDEEFTAQMITITPPDQDDSM
                     ECVDSERRPHFPQFSYSASGTA"
     protein_bind    complement(2112..2211)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2228..2247)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2229..2247)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2252..2268)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2252..2268)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2699..3490
                     /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      3583..4171
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4072..4091
                     /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 atgacaactt 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 cctgaagtgg 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 tgcccaactt tcttgtacaa agttggcatt ataagaaagc attgcttatc
     2161 aatttgttgc aacgaacagg tcactatcag tcaaaataaa atcattattt gccatccagc
     2221 tgatatcccc tatagtgagt cgtattacat ggtcatagct gtttcctggc agctctggcc
     2281 cgtgtctcaa aatctctgat gttacattgc acaagataaa aatatatcat catgcctcct
     2341 ctagaccagc caggacagaa atgcctcgac ttcgctgcta cccaaggttg ccgggtgacg
     2401 cacaccgtgg aaacggatga aggcacgaac ccagtggaca taagcctgtt cggttcgtaa
     2461 gctgtaatgc aagtagcgta tgcgctcacg caactggtcc agaaccttga ccgaacgcag
     2521 cggtggtaac ggcgcagtgg cggttttcat ggcttgttat gactgttttt ttggggtaca
     2581 gtctatgcct cgggcatcca agcagcaagc gcgttacgcc gtgggtcgat gtttgatgtt
     2641 atggagcagc aacgatgtta cgcagcaggg cagtcgccct aaaacaaagt taaacattat
     2701 gagggaagcg gtgatcgccg aagtatcgac tcaactatca gaggtagttg gcgtcatcga
     2761 gcgccatctc gaaccgacgt tgctggccgt acatttgtac ggctccgcag tggatggcgg
     2821 cctgaagcca cacagtgata ttgatttgct ggttacggtg accgtaaggc ttgatgaaac
     2881 aacgcggcga gctttgatca acgacctttt ggaaacttcg gcttcccctg gagagagcga
     2941 gattctccgc gctgtagaag tcaccattgt tgtgcacgac gacatcattc cgtggcgtta
     3001 tccagctaag cgcgaactgc aatttggaga atggcagcgc aatgacattc ttgcaggtat
     3061 cttcgagcca gccacgatcg acattgatct ggctatcttg ctgacaaaag caagagaaca
     3121 tagcgttgcc ttggtaggtc cagcggcgga ggaactcttt gatccggttc ctgaacagga
     3181 tctatttgag gcgctaaatg aaaccttaac gctatggaac tcgccgcccg actgggctgg
     3241 cgatgagcga aatgtagtgc ttacgttgtc ccgcatttgg tacagcgcag taaccggcaa
     3301 aatcgcgccg aaggatgtcg ctgccgactg ggcaatggag cgcctgccgg cccagtatca
     3361 gcccgtcata cttgaagcta gacaggctta tcttggacaa gaagaagatc gcttggcctc
     3421 gcgcgcagat cagttggaag aatttgtcca ctacgtgaaa ggcgagatca ccaaggtagt
     3481 cggcaaataa ccctcgagcc acccatgacc aaaatccctt aacgtgagtt acgcgtcgtt
     3541 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     3601 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     3661 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     3721 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     3781 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     3841 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     3901 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     3961 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     4021 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     4081 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     4141 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     4201 cctggccttt tgctggcctt ttgctcacat gtt
//
