LOCUS Exported 6126 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses eGFP-tagged human Akt2(WT). ACCESSION . VERSION . KEYWORDS pEGFP-Akt2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6126) AUTHORS Ebner M, Lucic I, Leonard TA, Yudushkin I TITLE PI(3,4,5)P3 Engagement Restricts Akt Activity to Cellular Membranes. JOURNAL Mol Cell. 2017 Feb 2;65(3):416-431.e6. doi: 10.1016/j.molcel.2016.12.028. PUBMED 28157504 REFERENCE 2 (bases 1 to 6126) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6126 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 15..34 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 137..931 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(191..210) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 801..820 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(1119..1138) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 1163..1210 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 1539..2127 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2028..2047 /label=pBR322ori-F /note="pBR322 origin, forward primer" enhancer 2302..2605 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 2606..2809 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 2760..2780 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 2848..2857 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 2854..3570 /codon_start=1 /product="enhanced GFP with monomerizing A206K mutation (Zacharias et al., 2002)" /label=mEGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(2899..2920) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(3160..3179) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 3507..3528 /label=EGFP-C /note="EGFP, forward primer" polyA_signal 5155..5276 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5192..5211) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5246..5265 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(5283..5738) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5420..5441) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 5632..5651 /label=F1ori-R /note="F1 origin, reverse primer" promoter 5765..5869 /gene="bla" /label=AmpR promoter promoter 5871..102 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(5894..5914) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 6079..88 /label=SV40 ori /note="SV40 origin of replication" ORIGIN 1 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag 61 aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg 121 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg 181 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc 241 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg 301 tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt 361 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg 421 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat 481 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca 541 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca 601 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa 661 ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa 721 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc 781 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga 841 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc 901 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac 961 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg 1021 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc 1081 atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata 1141 ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg 1201 tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg 1261 agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt 1321 tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca 1381 ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag 1441 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac 1501 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc 1561 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat 1621 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat 1681 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct 1741 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt 1801 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg 1861 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta 1921 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg 1981 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg 2041 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc 2101 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg 2161 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat 2221 aaccgtatta ccgccatgca ttagttatta atagtaatca attacggggt cattagttca 2281 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc 2341 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat 2401 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt 2461 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc 2521 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta 2581 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg 2641 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt 2701 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac 2761 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctg gtttagtgaa 2821 ccgtcagatc cgctagcgct accggtcgcc accatggtga gcaagggcga ggagctgttc 2881 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc 2941 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc 3001 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg 3061 cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg 3121 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc 3181 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc 3241 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac 3301 aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc 3361 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc 3421 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc caagctgagc 3481 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg 3541 atcactctcg gcatggacga gctgtacaag tccggactca gatctcgagc tatgaatgag 3601 gtgtctgtca tcaaagaagg ctggctccac aagcgtggtg aatacatcaa gacctggagg 3661 ccacggtact tcctgctgaa gagcgacggc tccttcattg ggtacaagga gaggcccgag 3721 gcccctgatc agactctacc ccccttaaac aacttctccg tagcagaatg ccagctgatg 3781 aagaccgaga ggccgcgacc caacaccttt gtcatacgct gcctgcagtg gaccacagtc 3841 atcgagagga ccttccacgt ggattctcca gacgagaggg aggagtggat gcgggccatc 3901 cagatggtcg ccaacagcct caagcagcgg gccccaggcg aggaccccat ggactacaag 3961 tgtggctccc ccagtgactc ctccacgact gaggagatgg aagtggcggt cagcaaggca 4021 cgggctaaag tgaccatgaa tgacttcgac tatctcaaac tccttggcaa gggaaccttt 4081 ggcaaagtca tcctggtgcg ggagaaggcc actggccgct actacgccat gaagatcctg 4141 cggaaggaag tcatcattgc caaggatgaa gtcgctcaca cagtcaccga gagccgggtc 4201 ctccagaaca ccaggcaccc gttcctcact gcgctgaagt atgccttcca gacccacgac 4261 cgcctgtgct ttgtgatgga gtatgccaac gggggtgagc tgttcttcca cctgtcccgg 4321 gagcgtgtct tcacagagga gcgggcccgg ttttatggtg cagagattgt ctcggctctt 4381 gagtacttgc actcgcggga cgtggtatac cgcgacatca agctggaaaa cctcatgctg 4441 gacaaagatg gccacatcaa gatcactgac tttggcctct gcaaagaggg catcagtgac 4501 ggggccacca tgaaaacctt ctgtgggacc ccggagtacc tggcgcctga ggtgctggag 4561 gacaatgact atggccgggc cgtggactgg tgggggctgg gtgtggtcat gtacgagatg 4621 atgtgcggcc gcctgccctt ctacaaccag gaccacgagc gcctcttcga gctcatcctc 4681 atggaagaga tccgcttccc gcgcacgctc agccccgagg ccaagtccct gcttgctggg 4741 ctgcttaaga aggaccccaa gcagaggctt ggtggggggc ccagcgatgc caaggaggtc 4801 atggagcaca ggttcttcct cagcatcaac tggcaggacg tggtccagaa gaagctcctg 4861 ccacccttca aacctcaggt cacgtccgag gtcgacacaa ggtacttcga tgatgaattt 4921 accgcccagt ccatcacaat cacaccccct gaccgctatg acagcctggg cttactggag 4981 ctggaccagc ggacccactt cccccagttc tcctactcgg ccagcatccg cgagtgatct 5041 agataactga tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac 5101 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg 5161 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa 5221 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaacgc 5281 gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc 5341 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga 5401 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc 5461 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc 5521 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag 5581 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa 5641 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac 5701 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa 5761 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat 5821 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga 5881 aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca 5941 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca 6001 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc 6061 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc 6121 catggc //