LOCUS Exported 4233 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_p.E17K 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 12, 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 92..109 /label=L4440 /note="L4440 vector, forward primer" terminator 268..295 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" terminator 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="MSDVAIVKEGWLHKRGKYIKTWRPRYFLLKNDGTFIGYKERPQDV DQREAPLNNFSVAQCQLMKTERPRPNTFIIRCLQWTTVIERTFHVETPEEREEWTTAIQ 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 atgccaactt tgtacaaaaa 661 agttggcacc atgagcgacg tggctattgt gaaggagggt tggctgcaca aacgagggaa 721 gtacatcaag acctggcggc cacgctactt cctcctcaag aatgatggca ccttcattgg 781 ctacaaggag cggccgcagg atgtggacca acgtgaggct cccctcaaca acttctctgt 841 ggcgcagtgc cagctgatga agacggagcg gccccggccc aacaccttca tcatccgctg 901 cctgcagtgg 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 //