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.D323Y
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4233)
  AUTHORS   Berger AH, Brooks AN, Wu X, Shrestha Y, Chouinard C, Piccioni F, 
            Bagul M, Kamburov A, Imielinski M, Hogstrom L, Zhu C, Yang X, Pantel
            S, Sakai R, Watson J, Kaplan N, Campbell JD, Singh S, Root DE, 
            Narayan R, Natoli T, Lahr DL, Tirosh I, Tamayo P, Getz G, Wong B, 
            Doench J, Subramanian A, Golub TR, Meyerson M, Boehm JS
  TITLE     High-throughput Phenotyping of Lung Cancer Somatic Mutations.
  JOURNAL   Cancer Cell. 2016 Aug 8;30(2):214-28. doi: 
            10.1016/j.ccell.2016.06.022. Epub 2016 Jul 28.
  PUBMED    27478040
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             189..1628
                     /codon_start=1
                     /gene="AKT1"
                     /product="human serine/threonine protein kinase activated 
                     by growth factors"
                     /label=Akt1
                     /translation="MSDVAIVKEGWLHKRGEYIKTWRPRYFLLKNDGTFIGYKERPQDV
                     DQREAPLNNFSVAQCQLMKTERPRPNTFIIRCLQWTTVIERTFHVETPEEREEWTTAIQ
                     TVADGLKKQEEEEMDFRSGSPSDNSGAEEMEVSLAKPKHRVTMNEFEYLKLLGKGTFGK
                     VILVKEKATGRYYAMKILKKEVIVAKDEVAHTLTENRVLQNSRHPFLTALKYSFQTHDR
                     LCFVMEYANGGELFFHLSRERVFSEDRARFYGAEIVSALDYLHSEKNVVYRDLKLENLM
                     LDKDGHIKITDFGLCKEGIKDGATMKTFCGTPEYLAPEVLEYNDYGRAVDWWGLGVVMY
                     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 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 gaggaggaga
      541 tggacttccg gtcgggctca cccagtgaca actcaggggc tgaagagatg gaggtgtccc
      601 tggccaagcc caagcaccgc gtgaccatga acgagtttga gtacctgaag ctgctgggca
      661 agggcacttt cggcaaggtg atcctggtga aggagaaggc cacaggccgc tactacgcca
      721 tgaagatcct caagaaggaa gtcatcgtgg ccaaggacga ggtggcccac acactcaccg
      781 agaaccgcgt cctgcagaac tccaggcacc ccttcctcac agccctgaag tactctttcc
      841 agacccacga ccgcctctgc tttgtcatgg agtacgccaa cgggggcgag ctgttcttcc
      901 acctgtcccg ggagcgtgtg ttctccgagg accgggcccg cttctatggc gctgagattg
      961 tgtcagccct ggactacctg cactcggaga agaacgtggt gtaccgggac ctcaagctgg
     1021 agaacctcat gctggacaag gacgggcaca ttaagatcac agacttcggg ctgtgcaagg
     1081 aggggatcaa ggacggtgcc accatgaaga ccttttgcgg cacacctgag tacctggccc
     1141 ccgaggtgct ggagtacaat gactacggcc gtgcagtgga ctggtggggg ctgggcgtgg
     1201 tcatgtacga gatgatgtgc ggtcgcctgc ccttctacaa ccaggaccat gagaagcttt
     1261 ttgagctcat cctcatggag gagatccgct tcccgcgcac gcttggtccc gaggccaagt
     1321 ccttgctttc agggctgctc aagaaggacc ccaagcagag gcttggcggg ggctccgagg
     1381 acgccaagga gatcatgcag catcgcttct ttgccggtat cgtgtggcag cacgtgtacg
     1441 agaagaagct cagcccaccc ttcaagcccc aggtcacgtc ggagactgac accaggtatt
     1501 ttgatgagga gttcacggcc cagatgatca ccatcacacc acctgaccaa gatgacagca
     1561 tggagtgtgt ggacagcgag cgcaggcccc acttccccca gttctcctac tcggccagcg
     1621 gcacggcctg attgccaact 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
//
