LOCUS       Exported                4230 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Gateway Donor vector containing AKT1, part of the Target Accelerator
            Plasmid Collection..
ACCESSION   .
VERSION     .
KEYWORDS    pDONR223_AKT1_WT
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 May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4230
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      34..622
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     523..542
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     776..793
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      952..979
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      1071..1157
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     1206..1228
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1220..1237
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1221..1237
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1253..1352
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             1352..2791
                     /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(2793..2892)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2909..2928)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2910..2928)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2933..2949)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2933..2949)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3380..4171
                     /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 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
       61 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
      121 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
      181 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
      241 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
      301 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
      361 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
      421 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
      481 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
      541 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
      601 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
      661 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
      721 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
      781 gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
      841 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg
      901 caacgcaatt aatacgcgta ccgctagcca ggaagagttt gtagaaacgc aaaaaggcca
      961 tccgtcagga tggccttctg cttagtttga tgcctggcag tttatggcgg gcgtcctgcc
     1021 cgccaccctc cgggccgttg cttcacaacg ttcaaatccg ctcccggcgg atttgtccta
     1081 ctcaggagag cgttcaccga caaacaacag ataaaacgaa aggcccagtc ttccgactga
     1141 gcctttcgtt ttatttgatg cctggcagtt ccctactctc gcgttaacgc tagcatggat
     1201 gttttcccag tcacgacgtt gtaaaacgac ggccagtctt aagctcgggc cccaaataat
     1261 gattttattt tgactgatag tgacctgttc gttgcaacaa attgatgagc aatgcttttt
     1321 tataatgcca actttgtaca aaaaagttgg catgagcgac gtggctattg tgaaggaggg
     1381 ttggctgcac aaacgagggg agtacatcaa gacctggcgg ccacgctact tcctcctcaa
     1441 gaatgatggc accttcattg gctacaagga gcggccgcag gatgtggacc aacgtgaggc
     1501 tcccctcaac aacttctctg tggcgcagtg ccagctgatg aagacggagc ggccccggcc
     1561 caacaccttc atcatccgct gcctgcagtg gaccactgtc atcgaacgca ccttccatgt
     1621 ggagactcct gaggagcggg aggagtggac aaccgccatc cagactgtgg ctgacggcct
     1681 caagaagcag gaggaggagg agatggactt ccggtcgggc tcacccagtg acaactcagg
     1741 ggctgaagag atggaggtgt ccctggccaa gcccaagcac cgcgtgacca tgaacgagtt
     1801 tgagtacctg aagctgctgg gcaagggcac tttcggcaag gtgatcctgg tgaaggagaa
     1861 ggccacaggc cgctactacg ccatgaagat cctcaagaag gaagtcatcg tggccaagga
     1921 cgaggtggcc cacacactca ccgagaaccg cgtcctgcag aactccaggc accccttcct
     1981 cacagccctg aagtactctt tccagaccca cgaccgcctc tgctttgtca tggagtacgc
     2041 caacgggggc gagctgttct tccacctgtc ccgggagcgt gtgttctccg aggaccgggc
     2101 ccgcttctat ggcgctgaga ttgtgtcagc cctggactac ctgcactcgg agaagaacgt
     2161 ggtgtaccgg gacctcaagc tggagaacct catgctggac aaggacgggc acattaagat
     2221 cacagacttc gggctgtgca aggaggggat caaggacggt gccaccatga agaccttttg
     2281 cggcacacct gagtacctgg cccccgaggt gctggaggac aatgactacg gccgtgcagt
     2341 ggactggtgg gggctgggcg tggtcatgta cgagatgatg tgcggtcgcc tgcccttcta
     2401 caaccaggac catgagaagc tttttgagct catcctcatg gaggagatcc gcttcccgcg
     2461 cacgcttggt cccgaggcca agtccttgct ttcagggctg ctcaagaagg accccaagca
     2521 gaggcttggc gggggctccg aggacgccaa ggagatcatg cagcatcgct tctttgccgg
     2581 tatcgtgtgg cagcacgtgt acgagaagaa gctcagccca cccttcaagc cccaggtcac
     2641 gtcggagact gacaccaggt attttgatga ggagttcacg gcccagatga tcaccatcac
     2701 accacctgac caagatgaca gcatggagtg tgtggacagc gagcgcaggc cccacttccc
     2761 ccagttctcc tactcggcca gcggcacggc ctgcccaact ttcttgtaca aagttggcat
     2821 tataagaaag cattgcttat caatttgttg caacgaacag gtcactatca gtcaaaataa
     2881 aatcattatt tgccatccag ctgatatccc ctatagtgag tcgtattaca tggtcatagc
     2941 tgtttcctgg cagctctggc ccgtgtctca aaatctctga tgttacattg cacaagataa
     3001 aaatatatca tcatgcctcc tctagaccag ccaggacaga aatgcctcga cttcgctgct
     3061 acccaaggtt gccgggtgac gcacaccgtg gaaacggatg aaggcacgaa cccagtggac
     3121 ataagcctgt tcggttcgta agctgtaatg caagtagcgt atgcgctcac gcaactggtc
     3181 cagaaccttg accgaacgca gcggtggtaa cggcgcagtg gcggttttca tggcttgtta
     3241 tgactgtttt tttggggtac agtctatgcc tcgggcatcc aagcagcaag cgcgttacgc
     3301 cgtgggtcga tgtttgatgt tatggagcag caacgatgtt acgcagcagg gcagtcgccc
     3361 taaaacaaag ttaaacatta tgagggaagc ggtgatcgcc gaagtatcga ctcaactatc
     3421 agaggtagtt ggcgtcatcg agcgccatct cgaaccgacg ttgctggccg tacatttgta
     3481 cggctccgca gtggatggcg gcctgaagcc acacagtgat attgatttgc tggttacggt
     3541 gaccgtaagg cttgatgaaa caacgcggcg agctttgatc aacgaccttt tggaaacttc
     3601 ggcttcccct ggagagagcg agattctccg cgctgtagaa gtcaccattg ttgtgcacga
     3661 cgacatcatt ccgtggcgtt atccagctaa gcgcgaactg caatttggag aatggcagcg
     3721 caatgacatt cttgcaggta tcttcgagcc agccacgatc gacattgatc tggctatctt
     3781 gctgacaaaa gcaagagaac atagcgttgc cttggtaggt ccagcggcgg aggaactctt
     3841 tgatccggtt cctgaacagg atctatttga ggcgctaaat gaaaccttaa cgctatggaa
     3901 ctcgccgccc gactgggctg gcgatgagcg aaatgtagtg cttacgttgt cccgcatttg
     3961 gtacagcgca gtaaccggca aaatcgcgcc gaaggatgtc gctgccgact gggcaatgga
     4021 gcgcctgccg gcccagtatc agcccgtcat acttgaagct agacaggctt atcttggaca
     4081 agaagaagat cgcttggcct cgcgcgcaga tcagttggaa gaatttgtcc actacgtgaa
     4141 aggcgagatc accaaggtag tcggcaaata accctcgagc cacccatgac caaaatccct
     4201 taacgtgagt tacgcgtcgt tccactgagc
//
