LOCUS Exported 9811 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Mammalian expression vector containing PIK3C2A tagged with GFP. ACCESSION . VERSION . KEYWORDS GFP-PIK3C2A SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9811) AUTHORS Posor Y, Eichhorn-Gruenig M, Puchkov D, Schoneberg J, Ullrich A, Lampe A, Muller R, Zarbakhsh S, Gulluni F, Hirsch E, Krauss M, Schultz C, Schmoranzer J, Noe F, Haucke V TITLE Spatiotemporal control of endocytosis by phosphatidylinositol-3,4-bisphosphate. JOURNAL Nature. 2013 Jul 11;499(7457):233-7. doi: 10.1038/nature12360. Epub 2013 Jul 3. PUBMED 23823722 REFERENCE 2 (bases 1 to 9811) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9811 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 134..437 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 438..641 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 592..612 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 680..689 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 686..1402 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 686..1402 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(731..752) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(992..1011) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1339..1360 /label=EGFP-C /note="EGFP, forward primer" polyA_signal 6672..6793 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(6709..6728) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 6763..6782 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(6800..7255) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(6937..6958) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 7149..7168 /label=F1ori-R /note="F1 origin, reverse primer" promoter 7282..7386 /gene="bla" /label=AmpR promoter promoter 7388..7745 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(7411..7431) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 7596..7731 /label=SV40 ori /note="SV40 origin of replication" primer_bind 7658..7677 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 7780..8574 /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(7834..7853) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 8444..8463 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(8762..8781) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 8806..8853 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 9182..9770 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 9671..9690 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat 61 taccgccatg cattagttat taatagtaat caattacggg gtcattagtt catagcccat 121 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg 181 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt 241 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag 301 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc 361 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag 421 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt 481 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc 541 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg 601 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tggtttagtg aaccgtcaga 661 tccgctagcg ctaccggtcg ccaccatggt gagcaagggc gaggagctgt tcaccggggt 721 ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg 781 cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg 841 caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt 901 cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg 961 ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga 1021 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa 1081 ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta 1141 tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat 1201 cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg 1261 ccccgtgctg ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc 1321 caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct 1381 cggcatggac gagctgtaca agtactcaga tctcgagctc caccgcggtg gcggccgctc 1441 tagaactagt atggctcaga tatttagcaa cagcggattt aaagaatgtc catcttcaca 1501 tccggaacca acaagagcaa aagatgtgga caaagaagaa gcattacaga tggaagcaga 1561 ggctttagca aaactgcaaa aggatagaca agtgactgac aatcagagag gctttgagtt 1621 gtcaagcagc accagaaaaa aagcacaggt ttataacaag caggattatg atctcatggt 1681 gtttcctgaa tcagattccc aaaaaagagc attagatatt gatgtagaaa agctcaccca 1741 agctgaactt gagaaactat tgctggatga cagtttcgag actaaaaaaa cacctgtatt 1801 accagttact cctattctga gcccttcctt ttcagcacag ctctatttta gacctactat 1861 tcagagagga cagtggccac ctggattacc tgggccttcc acttatgctt taccttctat 1921 ttatccttct acttacagta aacaggctgc attccaaaat ggcttcaatc caagaatgcc 1981 cacttttcca tctacagaac ctatatattt aagtcttccg ggacaatctc catatttctc 2041 atatcctttg acacctgcca caccctttca tccacaagga agcttaccta tctatcgtcc 2101 agtagtcagt actgacatgg caaaactatt tgacaaaata gctagtacat cagaattttt 2161 aaaaaatggg aaagcaagga ctgatttgga gataacagat tcaaaagtca gcaatctaca 2221 ggtatctcca aagtctgagg atatcagtaa atttgactgg ttagacttgg atcctctaag 2281 taagcctaag gtggataatg tggaggtatt agaccatgag gaagagaaaa atgtttcaag 2341 tttgctagca aaggatcctt gggatgctgt tcttcttgaa gagagatcga cagcaaattg 2401 tcatcttgaa agaaaggtga atggaaaatc cctttctgtg gcaactgtta caagaagcca 2461 gtctttaaat attcgaacaa ctcagcttgc aaaagcccag ggccatatat ctcagaaaga 2521 cccaaatggg accagtagtt tgccaactgg aagttctctt cttcaagaag ttgaagtaca 2581 gaatgaggag atggcagctt tttgtcgatc cattacaaaa ttgaagacca aatttccata 2641 taccaatcac cgcacaaacc caggctattt gttaagtcca gtcacagcgc aaagaaacat 2701 atgcggagaa aatgctagtg tgaaggtctc cattgacatt gaaggatttc agctaccagt 2761 tacttttacg tgtgatgtga gttctactgt agaaatcatt ataatgcaag ccctttgctg 2821 ggtacatgat gacttgaatc aagtagatgt tggcagctat gttctaaaag tttgtggtca 2881 agaggaagtg ctgcagaata atcattgcct tggaagtcat gagcatattc aaaactgtcg 2941 aaaatgggac acagaaatta gactacaact cttgaccttc agtgcaatgt gtcaaaatct 3001 ggcccgaaca gcagaagatg atgaaacacc cgtggattta aacaaacacc tgtatcaaat 3061 agaaaaacct tgcaaagaag ccatgacgag acaccctgtt gaagaactct tagattctta 3121 tcacaaccaa gtagaactgg ctcttcaaat tgaaaaccaa caccgagcag tagatcaagt 3181 aattaaagct gtaagaaaaa tctgtagtgc tttagatggt gtcgagactc ttgccattac 3241 agaatcagta aagaagctaa agagagcagt taatcttcca aggagtaaaa ctgctgatgt 3301 gacttctttg tttggaggag aagacactag caggagttca actaggggct cacttaatcc 3361 tgaaaatcct gttcaagtaa gcataaacca attaactgca gcaatttatg atcttctcag 3421 actccatgca aattctggta ggagtcctac agactgtgcc caaagtagca agagtgtcaa 3481 ggaagcatgg actacaacag agcagctcca gtttactatt tttgctgctc atggaatttc 3541 aagtaattgg gtatcaaatt atgaaaaata ctacttgata tgttcactgt ctcacaatgg 3601 aaaggatctt tttaaaccta ttcaatcaaa gaaggttggc acttacaaga atttcttcta 3661 tcttattaaa tgggatgaac taatcatttt tcctatccag atatcacaat tgccattaga 3721 atcagttctt caccttactc tttttggaat tttaaatcag agcagtggaa gttcccctga 3781 ttctaataag cagagaaagg gaccagaagc tttgggcaaa gtttctttac ctctttttga 3841 ctttaaacgg tttttaacat gtggaactaa acttctatat ctttggactt catcacatac 3901 aaattctgtt cctggaacag ttaccaaaaa aggatatgtc atggaaagaa tagtgctaca 3961 ggttgatttt ccttctcctg catttgatat tatttataca actcctcaag ttgacagaag 4021 cattatacag caacataact tagaaacact agagaatgat ataaaaggga aacttcttga 4081 tattcttcat aaagactcat cacttggact ttctaaagaa gataaagctt ttttatggga 4141 gaaacgttat tattgcttca aacacccaaa ttgtcttcct aaaatattag caagcgcccc 4201 aaactggaaa tgggttaatc ttgccaaaac ttactcattg cttcaccagt ggcctgcatt 4261 gtacccacta attgcattgg aacttcttga ttcaaaattt gctgatcagg aagtaagatc 4321 cctagctgtg acctggattg aggccattag tgatgatgag ctaacagatc ttcttccaca 4381 gtttgtacaa gctttgaaat atgaaattta cttgaatagt tcattagtgc aattcctttt 4441 gtccagggca ttgggaaata tccagatagc acacaattta tattggcttc tcaaagatgc 4501 cctgcatgat gtacagttta gtacccgata cgaacatgtt ttgggtgctc tcctgtcagt 4561 aggaggaaaa cgacttagag aagaacttct aaaacagacg aaacttgtac agcttttagg 4621 aggagtagca gaaaaagtaa ggcaggctag tggatcagcc agacaggttg ttctccaaag 4681 aagtatggaa cgagtacagt ccttttttca gaaaaataaa tgccgtctcc ctctcaagcc 4741 aagtctagtg gcaaaagaat taaatattaa gtcgtgttcc ttcttcagtt ctaatgctgt 4801 ccccctaaaa gtcacaatgg tgaatgctga ccctatggga gaagaaatta atgtcatgtt 4861 taaggttggt gaagatcttc ggcaagatat gttagcttta cagatgataa agattatgga 4921 taagatctgg cttaaagaag gactagatct gaggatggta attttcaaat gtctctcaac 4981 tggcagagat cgaggcatgg tggagctggt tcctgcttcc gataccctca ggaaaatcca 5041 agtggaatat ggtgtgacag gatcctttaa agataaacca cttgcagagt ggctaaggaa 5101 atacaatccc tctgaagaag aatatgaaaa ggcttcagag aactttatct attcctgtgc 5161 tggatgctgt gtagccacct atgttttagg catctgtgat cgacacaatg acaatataat 5221 gcttcgaagc acgggacaca tgtttcacat tgactttgga aagtttttgg gacatgcaca 5281 gatgtttggc agcttcaaaa gggatcgggc tccttttgtg ctgacctctg atatggcata 5341 tgtcattaat gggggtgaaa agcccaccat tcgttttcag ttgtttgtgg acctctgctg 5401 tcaggcctac aacttgataa gaaagcagac aaaccttttt cttaacctcc tttcactgat 5461 gattccttca gggttaccag aacttacaag tattcaagat ttgaaatacg ttagagatgc 5521 acttcaaccc caaactacag acgcagaagc tacaattttc tttactaggc ttattgaatc 5581 aagtttggga agcattgcca caaagtttaa cttcttcatt cacaaccttg ctcagcttcg 5641 tttttctggt cttccttcta atgatgagcc catcctttca ttttcaccta aaacatactc 5701 ctttagacaa gatggtcgaa tcaaggaagt ctctgttttt acatatcata agaaatacaa 5761 cccagataaa cattatattt atgtagtccg aattttgagg gaaggacaga ttgaaccatc 5821 atttgtcttc cgaacatttg acgaatttca ggaacttcac aataagctca gtattatttt 5881 tccactttgg aagttaccag gctttcctaa taggatggtt ctaggaagaa cacacataaa 5941 agatgtagca gccaaaagga aaattgagtt aaacagttac ttacagagtt tgatgaatgc 6001 ttcaacggat gtagcagagt gtgatcttgt ttgtactttc ttccaccctt tacttcgtga 6061 tgagaaagct gaagggatag ctaggtctgc agatgcaggt tccttcagtc ctactccagg 6121 ccaaatagga ggagctgtga aattatccat ctcttaccga aatggtactc ttttcatcat 6181 ggtgatgcat atcaaagatc ttgttactga agatggagct gacccaaatc catatgtcaa 6241 aacataccta cttccagata accacaaaac atccaaacgt aaaaccaaaa tttcacgaaa 6301 aacgaggaat ccgacattca atgaaatgct tgtatacagt ggatatagca aagaaaccct 6361 aagacagcga gaacttcaac taagtgtact cagtgcagaa tctctgcggg agaatttttt 6421 cttgggtgga gtaaccctgc ctttgaaaga tttcaacttg agcaaagaga cggttaaatg 6481 gtatcagctg actgcggcaa catacttgta agaattctgc agtcgacggt accgcgggcc 6541 cgggatccac cggatctaga taactgatca taatcagcca taccacattt gtagaggttt 6601 tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa 6661 ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca 6721 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca 6781 tcaatgtatc ttaacgcgta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat 6841 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa 6901 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta 6961 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca 7021 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat 7081 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg 7141 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc 7201 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt 7261 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca 7321 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg 7381 aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt 7441 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca 7501 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt 7561 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt 7621 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg 7681 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt 7741 gcaaagatcg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg 7801 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag 7861 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt 7921 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc agcgcggcta 7981 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg 8041 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt 8101 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat 8161 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg 8221 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca 8281 gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc 8341 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc 8401 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat 8461 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc 8521 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga 8581 ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt 8641 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga 8701 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccctagg gggaggctaa 8761 ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac 8821 agaataaaac gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct 8881 ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct 8941 tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg 9001 cggcaggccc tgccatagcc tcaggttact catatatact ttagattgat ttaaaacttc 9061 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc 9121 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt 9181 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac 9241 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct 9301 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact 9361 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg 9421 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata 9481 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga 9541 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag 9601 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg 9661 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac 9721 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca 9781 acgcggcctt tttacggttc ctggcctttt g //