LOCUS       Exported                4230 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.R370C
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 13, 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"
     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             189..1625
                     /codon_start=1
                     /gene="AKT1"
                     /product="human serine/threonine protein kinase activated 
                     by growth factors"
                     /label=Akt1
                     /translation="MSDVAIVKEGWLHKRGEYIKTWRPRYFLLKNDGTFIGYKERPQDV
                     DQREAPLNNFSVAQCQLMKTERPRPNTFIIRCLQWTTVIERTFHVETPEEREEWTTAIQ
                     TVADGLKKQEEEMDFRSGSPSDNSGAEEMEVSLAKPKHRVTMNEFEYLKLLGKGTFGKV
                     ILVKEKATGRYYAMKILKKEVIVAKDEVAHTLTENRVLQNSRHPFLTALKYSFQTHDRL
                     CFVMEYANGGELFFHLSRERVFSEDRARFYGAEIVSALDYLHSEKNVVYRDLKLENLML
                     DKDGHIKITDFGLCKEGIKDGATMKTFCGTPEYLAPEVLEDNDYGRAVDWWGLGVVMYE
                     MMCGRLPFYNQDHEKLFELILMEEIRFPCTLGPEAKSLLSGLLKKDPKQRLGGGSEDAK
                     EIMQHRFFAGIVWQHVYEKKLSPPFKPQVTSETDTRYFDEEFTAQMITITPPDQDDSME
                     CVDSERRPHFPQFSYSASGTA"
     protein_bind    complement(1630..1729)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1746..1765)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1747..1765)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1770..1786)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1770..1786)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2217..3008
                     /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      3101..3689
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3590..3609
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3843..3860
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      4019..4046
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      4138..4224
                     /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 aagttggcat gagcgacgtg gctattgtga aggagggttg gctgcacaaa cgaggggagt
      241 acatcaagac ctggcggcca cgctacttcc tcctcaagaa tgatggcacc ttcattggct
      301 acaaggagcg gccgcaggat gtggaccaac gtgaggctcc cctcaacaac ttctctgtgg
      361 cgcagtgcca gctgatgaag acggagcggc cccggcccaa caccttcatc atccgctgcc
      421 tgcagtggac cactgtcatc gaacgcacct tccatgtgga gactcctgag gagcgggagg
      481 agtggacaac cgccatccag actgtggctg acggcctcaa gaagcaggag gaggagatgg
      541 acttccggtc gggctcaccc agtgacaact caggggctga agagatggag gtgtccctgg
      601 ccaagcccaa gcaccgcgtg accatgaacg agtttgagta cctgaagctg ctgggcaagg
      661 gcactttcgg caaggtgatc ctggtgaagg agaaggccac aggccgctac tacgccatga
      721 agatcctcaa gaaggaagtc atcgtggcca aggacgaggt ggcccacaca ctcaccgaga
      781 accgcgtcct gcagaactcc aggcacccct tcctcacagc cctgaagtac tctttccaga
      841 cccacgaccg cctctgcttt gtcatggagt acgccaacgg gggcgagctg ttcttccacc
      901 tgtcccggga gcgtgtgttc tccgaggacc gggcccgctt ctatggcgct gagattgtgt
      961 cagccctgga ctacctgcac tcggagaaga acgtggtgta ccgggacctc aagctggaga
     1021 acctcatgct ggacaaggac gggcacatta agatcacaga cttcgggctg tgcaaggagg
     1081 ggatcaagga cggtgccacc atgaagacct tttgcggcac acctgagtac ctggcccccg
     1141 aggtgctgga ggacaatgac tacggccgtg cagtggactg gtgggggctg ggcgtggtca
     1201 tgtacgagat gatgtgcggt cgcctgccct tctacaacca ggaccatgag aagctttttg
     1261 agctcatcct catggaggag atccgcttcc cgtgcacgct tggtcccgag gccaagtcct
     1321 tgctttcagg gctgctcaag aaggacccca agcagaggct tggcgggggc tccgaggacg
     1381 ccaaggagat catgcagcat cgcttctttg ccggtatcgt gtggcagcac gtgtacgaga
     1441 agaagctcag cccacccttc aagccccagg tcacgtcgga gactgacacc aggtattttg
     1501 atgaggagtt cacggcccag atgatcacca tcacaccacc tgaccaagat gacagcatgg
     1561 agtgtgtgga cagcgagcgc aggccccact tcccccagtt ctcctactcg gccagcggca
     1621 cggcctgatt gccaactttc ttgtacaaag ttggcattat aagaaagcat tgcttatcaa
     1681 tttgttgcaa cgaacaggtc actatcagtc aaaataaaat cattatttgc catccagctg
     1741 atatccccta tagtgagtcg tattacatgg tcatagctgt ttcctggcag ctctggcccg
     1801 tgtctcaaaa tctctgatgt tacattgcac aagataaaaa tatatcatca tgcctcctct
     1861 agaccagcca ggacagaaat gcctcgactt cgctgctacc caaggttgcc gggtgacgca
     1921 caccgtggaa acggatgaag gcacgaaccc agtggacata agcctgttcg gttcgtaagc
     1981 tgtaatgcaa gtagcgtatg cgctcacgca actggtccag aaccttgacc gaacgcagcg
     2041 gtggtaacgg cgcagtggcg gttttcatgg cttgttatga ctgttttttt ggggtacagt
     2101 ctatgcctcg ggcatccaag cagcaagcgc gttacgccgt gggtcgatgt ttgatgttat
     2161 ggagcagcaa cgatgttacg cagcagggca gtcgccctaa aacaaagtta aacattatga
     2221 gggaagcggt gatcgccgaa gtatcgactc aactatcaga ggtagttggc gtcatcgagc
     2281 gccatctcga accgacgttg ctggccgtac atttgtacgg ctccgcagtg gatggcggcc
     2341 tgaagccaca cagtgatatt gatttgctgg ttacggtgac cgtaaggctt gatgaaacaa
     2401 cgcggcgagc tttgatcaac gaccttttgg aaacttcggc ttcccctgga gagagcgaga
     2461 ttctccgcgc tgtagaagtc accattgttg tgcacgacga catcattccg tggcgttatc
     2521 cagctaagcg cgaactgcaa tttggagaat ggcagcgcaa tgacattctt gcaggtatct
     2581 tcgagccagc cacgatcgac attgatctgg ctatcttgct gacaaaagca agagaacata
     2641 gcgttgcctt ggtaggtcca gcggcggagg aactctttga tccggttcct gaacaggatc
     2701 tatttgaggc gctaaatgaa accttaacgc tatggaactc gccgcccgac tgggctggcg
     2761 atgagcgaaa tgtagtgctt acgttgtccc gcatttggta cagcgcagta accggcaaaa
     2821 tcgcgccgaa ggatgtcgct gccgactggg caatggagcg cctgccggcc cagtatcagc
     2881 ccgtcatact tgaagctaga caggcttatc ttggacaaga agaagatcgc ttggcctcgc
     2941 gcgcagatca gttggaagaa tttgtccact acgtgaaagg cgagatcacc aaggtagtcg
     3001 gcaaataacc ctcgagccac ccatgaccaa aatcccttaa cgtgagttac gcgtcgttcc
     3061 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     3121 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     3181 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     3241 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     3301 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     3361 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     3421 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     3481 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     3541 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     3601 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     3661 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     3721 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     3781 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     3841 gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg
     3901 cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca
     3961 gtgagcgcaa cgcaattaat acgcgtaccg ctagccagga agagtttgta gaaacgcaaa
     4021 aaggccatcc gtcaggatgg ccttctgctt agtttgatgc ctggcagttt atggcgggcg
     4081 tcctgcccgc caccctccgg gccgttgctt cacaacgttc aaatccgctc ccggcggatt
     4141 tgtcctactc aggagagcgt tcaccgacaa acaacagata aaacgaaagg cccagtcttc
     4201 cgactgagcc tttcgtttta tttgatgcct
//
