LOCUS Exported 7319 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Mammalian expression of GFP-tagged Dynamin protein.. ACCESSION . VERSION . KEYWORDS pEGFP-N1 Dynamin1 R290A D291A T292A SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7319) AUTHORS Kong L, Sochacki KA, Wang H, Fang S, Canagarajah B, Kehr AD, Rice WJ, Strub MP, Taraska JW, Hinshaw JE TITLE Cryo-EM of the dynamin polymer assembled on lipid membrane. JOURNAL Nature. 2018 Aug;560(7717):258-262. doi: 10.1038/s41586-018-0378-6. Epub 2018 Aug 1. PUBMED 30069048 REFERENCE 2 (bases 1 to 7319) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7319 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 61..364 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 365..568 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 519..539 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 3259..3268 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 3265..3984 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 3265..3981 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(3310..3331) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(3571..3590) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 3918..3939 /label=EGFP-C /note="EGFP, forward primer" polyA_signal 4107..4228 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4144..4163) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4198..4217 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(4235..4690) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4372..4393) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 4584..4603 /label=F1ori-R /note="F1 origin, reverse primer" promoter 4717..4821 /gene="bla" /label=AmpR promoter promoter 4823..5180 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(4846..4866) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 5031..5166 /label=SV40 ori /note="SV40 origin of replication" primer_bind 5093..5112 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 5215..6009 /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(5269..5288) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 5879..5898 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(6197..6216) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 6241..6288 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 6617..7205 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 7106..7125 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg 61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt 121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca 181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc 241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta 301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac 361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg 421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg 481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt 541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta 601 ccggactcag atcctgccat gggcaaccgc ggcatggaag atctcatccc gctggtcaac 661 cggctgcaag acgccttctc tgccatcggc cagaacgcgg acctcgacct gccgcagatc 721 gctgtggtgg gcggccagag cgccggcaag agctcggtgc tcgagaattt cgtaggcagg 781 gacttcttgc ctcgaggatc tggcattgtt acccgacgtc ccctggtctt gcagctggtc 841 aatgcaacca cagaatatgc cgagttcctg cactgcaagg gaaagaaatt caccgacttc 901 gaggaggtgc gccttgagat cgaggccgag accgacaggg tcaccggcac caacaagggc 961 atctcgccgg tgcctatcaa cctccgcgtc tactcgccgc acgtgctgaa cctgaccctg 1021 gtggacctgc ccggaatgac caaggtcccg gtgggggacc aacctcccga catcgagttc 1081 cagatccgag acatgcttat gcagtttgtc accaaggaga actgcctcat cctggccgtg 1141 tcccccgcca actctgacct ggccaattct gacgccctca aggtcgccaa ggaggtggac 1201 ccccagggcc agcgcaccat cggggtcatc accaagctgg acctgatgga cgagggcaca 1261 gatgcccgtg atgtgctgga gaacaagctg ctccccctgc gcagaggcta cattggagtg 1321 gtgaaccgga gccagaagga cattgatggc aagaaggaca ttaccgccgc cttggctgct 1381 gaacgaaagt tcttcctctc ccatccatct tatcgccact tggctgaccg tatgggcacg 1441 ccctacctgc agaaggtcct caatcagcaa ctgacgaacc acatcgccgc cgccctgccg 1501 gggctgcgga acaagctgca gagccagcta ctgtccattg agaaggaggt ggaggaatac 1561 aagaacttcc gccctgatga cccagctcgc aagaccaagg ccctgctgca gatggtccag 1621 cagtttgccg tagactttga gaagcgcatt gagggctcag gagatcagat cgacacctac 1681 gaactgtcag ggggagcccg cattaaccga atcttccacg agcgcttccc tttcgagctg 1741 gtcaagatgg agtttgatga gaaggaactc cgaagggaga tcagctatgc tatcaagaat 1801 atccatggca ttagaacggg gctgtttacc ccagacatgg cctttgagac cattgtgaaa 1861 aagcaggtga agaagatccg agaaccgtgt ctcaagtgtg tggacatggt tatctcggag 1921 ctaatcagca ccgttagaca gtgcaccaag aagctccagc agtacccgcg gctacgggag 1981 gagatggagc gcatcgtgac cacccacatc cgggagcgcg agggccgcac taaggagcag 2041 gtcatgcttc tcatcgatat cgagctggct tacatgaaca ccaaccatga ggacttcata 2101 ggctttgcca atgctcagca gaggagcaac cagatgaaca agaagaagac ttcagggaac 2161 caggatgaga ttctggtcat ccgcaagggc tggctgacta tcaataatat tggcatcatg 2221 aaagggggct ccaaggagta ctggtttgtg ctgactgctg agaatctgtc ctggtacaag 2281 gatgatgagg agaaagagaa gaaatacatg ctgtctgtgg acaacctcaa gctgcgggac 2341 gtggagaagg gctttatgtc gagcaagcat atctttgccc tctttaacac ggagcagagg 2401 aatgtctaca aggattatcg gcagctggag ctagcctgtg agacacagga ggaggtggac 2461 agctggaagg cctccttcct gagggctggc gtgtaccctg agcgtgttgg ggacaaagag 2521 aaagccagcg agaccgagga gaatggctcc gacagcttca tgcattccat ggacccacag 2581 ctggaacggc aagtggagac catccggaat cttgtggact catacatggc cattgtcaac 2641 aagaccgtga gggacctcat gcccaagacc atcatgcacc tcatgattaa caataccaag 2701 gagttcatct tctcggagct gctggccaac ctgtactcgt gtggggacca gaacacgctg 2761 atggaggagt cggcggagca ggcacagcgg cgcgacgaga tgctgcgcat gtaccacgca 2821 ctgaaggagg cgctcagcat catcggcaac atcaacacga ccaccgtcag cacgcccatg 2881 cccccgcccg tggacgactc ctggctgcag gtgcagagcg taccggccgg acgcaggtcg 2941 cccacgtcca gccccacgcc gcagcgccga gcccccgccg tgcccccagc ccggcccggg 3001 tcgcggggcc ctgctcctgg gcctccgcct gctgggtccg ccctgggggg ggcgcccccc 3061 gtgccctcca ggccgggggc ttcccctgac cctttcggcc ctccccctca ggtgccctcg 3121 cgccccaacc gcgccccgcc cggggtcccc agccgatcgg gtcaggcaag tccatcccgt 3181 cctgagagcc ccaggccccc cttcgacaag cttcgaattc tgcagtcgac ggtaccgcgg 3241 gcccgggatc caccggtcgc caccatggtg agcaagggcg aggagctgtt caccggggtg 3301 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc 3361 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc 3421 aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc 3481 agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc 3541 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag 3601 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag 3661 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat 3721 atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc 3781 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc 3841 cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc 3901 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc 3961 ggcatggacg agctgtacaa gtaaagcggc cgcgactcta gatcataatc agccatacca 4021 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac 4081 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat 4141 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg 4201 gtttgtccaa actcatcaat gtatcttaag gcgtaaattg taagcgttaa tattttgtta 4261 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc 4321 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg 4381 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat 4441 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc 4501 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag 4561 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg 4621 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta 4681 cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt 4741 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa 4801 taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt 4861 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca 4921 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa 4981 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc 5041 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg 5101 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg 5161 gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt 5221 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat 5281 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag 5341 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac 5401 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac 5461 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc 5521 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg 5581 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag 5641 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat 5701 caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag 5761 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc 5821 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg 5881 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg 5941 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag 6001 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat 6061 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc 6121 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc 6181 ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga 6241 cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg 6301 ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg 6361 cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg 6421 cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga 6481 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc 6541 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa 6601 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa 6661 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc 6721 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt 6781 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc 6841 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac 6901 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca 6961 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg 7021 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag 7081 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt 7141 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat 7201 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc 7261 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gccatgcat //