LOCUS       Exported                4233 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Gateway Donor vector containing AKT1 , part of the Target 
            Accelerator Plasmid Collection..
ACCESSION   .
VERSION     .
KEYWORDS    pDONR223_AKT1_p.E267G
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 May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4233
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     43..65
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     57..74
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     58..74
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    90..189
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             192..1631
                     /codon_start=1
                     /gene="AKT1"
                     /product="human serine/threonine protein kinase activated 
                     by growth factors"
                     /label=Akt1
                     /translation="MSDVAIVKEGWLHKRGEYIKTWRPRYFLLKNDGTFIGYKERPQDV
                     DQREAPLNNFSVAQCQLMKTERPRPNTFIIRCLQWTTVIERTFHVETPEEREEWTTAIQ
                     TVADGLKKQEEEEMDFRSGSPSDNSGAEEMEVSLAKPKHRVTMNEFEYLKLLGKGTFGK
                     VILVKEKATGRYYAMKILKKEVIVAKDEVAHTLTENRVLQNSRHPFLTALKYSFQTHDR
                     LCFVMEYANGGELFFHLSRERVFSEDRARFYGAEIVSALDYLHSGKNVVYRDLKLENLM
                     LDKDGHIKITDFGLCKEGIKDGATMKTFCGTPEYLAPEVLEDNDYGRAVDWWGLGVVMY
                     EMMCGRLPFYNQDHEKLFELILMEEIRFPRTLGPEAKSLLSGLLKKDPKQRLGGGSEDA
                     KEIMQHRFFAGIVWQHVYEKKLSPPFKPQVTSETDTRYFDEEFTAQMITITPPDQDDSM
                     ECVDSERRPHFPQFSYSASGTA"
     protein_bind    complement(1633..1732)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1749..1768)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1750..1768)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1773..1789)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1773..1789)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2220..3011
                     /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      3104..3692
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3593..3612
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3846..3863
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      4022..4049
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      4141..4227
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ggcagttccc tactctcgcg ttaacgctag catggatgtt ttcccagtca cgacgttgta
       61 aaacgacggc cagtcttaag ctcgggcccc aaataatgat tttattttga ctgatagtga
      121 cctgttcgtt gcaacaaatt gatgagcaat gcttttttat aatgccaact ttgtacaaaa
      181 aagttggcac catgagcgac gtggctattg tgaaggaggg ttggctgcac aaacgagggg
      241 agtacatcaa gacctggcgg ccacgctact tcctcctcaa gaatgatggc accttcattg
      301 gctacaagga gcggccgcag gatgtggacc aacgtgaggc tcccctcaac aacttctctg
      361 tggcgcagtg ccagctgatg aagacggagc ggccccggcc caacaccttc atcatccgct
      421 gcctgcagtg gaccactgtc atcgaacgca ccttccatgt ggagactcct gaggagcggg
      481 aggagtggac aaccgccatc cagactgtgg ctgacggcct caagaagcag gaggaggagg
      541 agatggactt ccggtcgggc tcacccagtg acaactcagg ggctgaagag atggaggtgt
      601 ccctggccaa gcccaagcac cgcgtgacca tgaacgagtt tgagtacctg aagctgctgg
      661 gcaagggcac tttcggcaag gtgatcctgg tgaaggagaa ggccacaggc cgctactacg
      721 ccatgaagat cctcaagaag gaagtcatcg tggccaagga cgaggtggcc cacacactca
      781 ccgagaaccg cgtcctgcag aactccaggc accccttcct cacagccctg aagtactctt
      841 tccagaccca cgaccgcctc tgctttgtca tggagtacgc caacgggggc gagctgttct
      901 tccacctgtc ccgggagcgt gtgttctccg aggaccgggc ccgcttctat ggcgctgaga
      961 ttgtgtcagc cctggactac ctgcactcgg ggaagaacgt ggtgtaccgg gacctcaagc
     1021 tggagaacct catgctggac aaggacgggc acattaagat cacagacttc gggctgtgca
     1081 aggaggggat caaggacggt gccaccatga agaccttttg cggcacacct gagtacctgg
     1141 cccccgaggt gctggaggac aatgactacg gccgtgcagt ggactggtgg gggctgggcg
     1201 tggtcatgta cgagatgatg tgcggtcgcc tgcccttcta caaccaggac catgagaagc
     1261 tttttgagct catcctcatg gaggagatcc gcttcccgcg cacgcttggt cccgaggcca
     1321 agtccttgct ttcagggctg ctcaagaagg accccaagca gaggcttggc gggggctccg
     1381 aggacgccaa ggagatcatg cagcatcgct tctttgccgg tatcgtgtgg cagcacgtgt
     1441 acgagaagaa gctcagccca cccttcaagc cccaggtcac gtcggagact gacaccaggt
     1501 attttgatga ggagttcacg gcccagatga tcaccatcac accacctgac caagatgaca
     1561 gcatggagtg tgtggacagc gagcgcaggc cccacttccc ccagttctcc tactcggcca
     1621 gcggcacggc ctgcccaact ttcttgtaca aagttggcat tataagaaag cattgcttat
     1681 caatttgttg caacgaacag gtcactatca gtcaaaataa aatcattatt tgccatccag
     1741 ctgatatccc ctatagtgag tcgtattaca tggtcatagc tgtttcctgg cagctctggc
     1801 ccgtgtctca aaatctctga tgttacattg cacaagataa aaatatatca tcatgcctcc
     1861 tctagaccag ccaggacaga aatgcctcga cttcgctgct acccaaggtt gccgggtgac
     1921 gcacaccgtg gaaacggatg aaggcacgaa cccagtggac ataagcctgt tcggttcgta
     1981 agctgtaatg caagtagcgt atgcgctcac gcaactggtc cagaaccttg accgaacgca
     2041 gcggtggtaa cggcgcagtg gcggttttca tggcttgtta tgactgtttt tttggggtac
     2101 agtctatgcc tcgggcatcc aagcagcaag cgcgttacgc cgtgggtcga tgtttgatgt
     2161 tatggagcag caacgatgtt acgcagcagg gcagtcgccc taaaacaaag ttaaacatta
     2221 tgagggaagc ggtgatcgcc gaagtatcga ctcaactatc agaggtagtt ggcgtcatcg
     2281 agcgccatct cgaaccgacg ttgctggccg tacatttgta cggctccgca gtggatggcg
     2341 gcctgaagcc acacagtgat attgatttgc tggttacggt gaccgtaagg cttgatgaaa
     2401 caacgcggcg agctttgatc aacgaccttt tggaaacttc ggcttcccct ggagagagcg
     2461 agattctccg cgctgtagaa gtcaccattg ttgtgcacga cgacatcatt ccgtggcgtt
     2521 atccagctaa gcgcgaactg caatttggag aatggcagcg caatgacatt cttgcaggta
     2581 tcttcgagcc agccacgatc gacattgatc tggctatctt gctgacaaaa gcaagagaac
     2641 atagcgttgc cttggtaggt ccagcggcgg aggaactctt tgatccggtt cctgaacagg
     2701 atctatttga ggcgctaaat gaaaccttaa cgctatggaa ctcgccgccc gactgggctg
     2761 gcgatgagcg aaatgtagtg cttacgttgt cccgcatttg gtacagcgca gtaaccggca
     2821 aaatcgcgcc gaaggatgtc gctgccgact gggcaatgga gcgcctgccg gcccagtatc
     2881 agcccgtcat acttgaagct agacaggctt atcttggaca agaagaagat cgcttggcct
     2941 cgcgcgcaga tcagttggaa gaatttgtcc actacgtgaa aggcgagatc accaaggtag
     3001 tcggcaaata accctcgagc cacccatgac caaaatccct taacgtgagt tacgcgtcgt
     3061 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     3121 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     3181 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     3241 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     3301 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     3361 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     3421 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     3481 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     3541 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     3601 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     3661 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     3721 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     3781 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     3841 agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc
     3901 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg
     3961 gcagtgagcg caacgcaatt aatacgcgta ccgctagcca ggaagagttt gtagaaacgc
     4021 aaaaaggcca tccgtcagga tggccttctg cttagtttga tgcctggcag tttatggcgg
     4081 gcgtcctgcc cgccaccctc cgggccgttg cttcacaacg ttcaaatccg ctcccggcgg
     4141 atttgtccta ctcaggagag cgttcaccga caaacaacag ataaaacgaa aggcccagtc
     4201 ttccgactga gcctttcgtt ttatttgatg cct
//
