LOCUS Exported 7319 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Mammalian expression plasmid of GFP-tagged Dynamin protein.. ACCESSION . VERSION . KEYWORDS pEGFP-N1 Dynamin1 D406A M407A 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 78..381 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 382..585 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 536..556 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 3276..3285 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 3282..4001 /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 3282..3998 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(3327..3348) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(3588..3607) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 3935..3956 /label=EGFP-C /note="EGFP, forward primer" polyA_signal 4124..4245 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4161..4180) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4215..4234 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(4252..4707) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4389..4410) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 4601..4620 /label=F1ori-R /note="F1 origin, reverse primer" promoter 4734..4838 /gene="bla" /label=AmpR promoter promoter 4840..5197 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(4863..4883) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 5048..5183 /label=SV40 ori /note="SV40 origin of replication" primer_bind 5110..5129 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 5232..6026 /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(5286..5305) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 5896..5915 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(6214..6233) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 6258..6305 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 6634..7222 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 7123..7142 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 gtattaccgc catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc 61 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc 121 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg 181 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat 241 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc 301 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta 361 ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag 421 cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt 481 tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa 541 atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt 601 cagatccgct agcgctaccg gactcagatc ctgccatggg caaccgcggc atggaagatc 661 tcatcccgct ggtcaaccgg ctgcaagacg ccttctctgc catcggccag aacgcggacc 721 tcgacctgcc gcagatcgct gtggtgggcg gccagagcgc cggcaagagc tcggtgctcg 781 agaatttcgt aggcagggac ttcttgcctc gaggatctgg cattgttacc cgacgtcccc 841 tggtcttgca gctggtcaat gcaaccacag aatatgccga gttcctgcac tgcaagggaa 901 agaaattcac cgacttcgag gaggtgcgcc ttgagatcga ggccgagacc gacagggtca 961 ccggcaccaa caagggcatc tcgccggtgc ctatcaacct ccgcgtctac tcgccgcacg 1021 tgctgaacct gaccctggtg gacctgcccg gaatgaccaa ggtcccggtg ggggaccaac 1081 ctcccgacat cgagttccag atccgagaca tgcttatgca gtttgtcacc aaggagaact 1141 gcctcatcct ggccgtgtcc cccgccaact ctgacctggc caattctgac gccctcaagg 1201 tcgccaagga ggtggacccc cagggccagc gcaccatcgg ggtcatcacc aagctggacc 1261 tgatggacga gggcacagat gcccgtgatg tgctggagaa caagctgctc cccctgcgca 1321 gaggctacat tggagtggtg aaccggagcc agaaggacat tgatggcaag aaggacatta 1381 ccgccgcctt ggctgctgaa cgaaagttct tcctctccca tccatcttat cgccacttgg 1441 ctgaccgtat gggcacgccc tacctgcaga aggtcctcaa tcagcaactg acgaaccaca 1501 tccgggacac actgccgggg ctgcggaaca agctgcagag ccagctactg tccattgaga 1561 aggaggtgga ggaatacaag aacttccgcc ctgatgaccc agctcgcaag accaaggccc 1621 tgctgcagat ggtccagcag tttgccgtag actttgagaa gcgcattgag ggctcaggag 1681 atcagatcga cacctacgaa ctgtcagggg gagcccgcat taaccgaatc ttccacgagc 1741 gcttcccttt cgagctggtc aagatggagt ttgatgagaa ggaactccga agggagatca 1801 gctatgctat caagaatatc catggcatta gaacggggct gtttacccca gccgccgcct 1861 ttgagaccat tgtgaaaaag caggtgaaga agatccgaga accgtgtctc aagtgtgtgg 1921 acatggttat ctcggagcta atcagcaccg ttagacagtg caccaagaag ctccagcagt 1981 acccgcggct acgggaggag atggagcgca tcgtgaccac ccacatccgg gagcgcgagg 2041 gccgcactaa ggagcaggtc atgcttctca tcgatatcga gctggcttac atgaacacca 2101 accatgagga cttcataggc tttgccaatg ctcagcagag gagcaaccag atgaacaaga 2161 agaagacttc agggaaccag gatgagattc tggtcatccg caagggctgg ctgactatca 2221 ataatattgg catcatgaaa gggggctcca aggagtactg gtttgtgctg actgctgaga 2281 atctgtcctg gtacaaggat gatgaggaga aagagaagaa atacatgctg tctgtggaca 2341 acctcaagct gcgggacgtg gagaagggct ttatgtcgag caagcatatc tttgccctct 2401 ttaacacgga gcagaggaat gtctacaagg attatcggca gctggagcta gcctgtgaga 2461 cacaggagga ggtggacagc tggaaggcct ccttcctgag ggctggcgtg taccctgagc 2521 gtgttgggga caaagagaaa gccagcgaga ccgaggagaa tggctccgac agcttcatgc 2581 attccatgga cccacagctg gaacggcaag tggagaccat ccggaatctt gtggactcat 2641 acatggccat tgtcaacaag accgtgaggg acctcatgcc caagaccatc atgcacctca 2701 tgattaacaa taccaaggag ttcatcttct cggagctgct ggccaacctg tactcgtgtg 2761 gggaccagaa cacgctgatg gaggagtcgg cggagcaggc acagcggcgc gacgagatgc 2821 tgcgcatgta ccacgcactg aaggaggcgc tcagcatcat cggcaacatc aacacgacca 2881 ccgtcagcac gcccatgccc ccgcccgtgg acgactcctg gctgcaggtg cagagcgtac 2941 cggccggacg caggtcgccc acgtccagcc ccacgccgca gcgccgagcc cccgccgtgc 3001 ccccagcccg gcccgggtcg cggggccctg ctcctgggcc tccgcctgct gggtccgccc 3061 tggggggggc gccccccgtg ccctccaggc cgggggcttc ccctgaccct ttcggccctc 3121 cccctcaggt gccctcgcgc cccaaccgcg ccccgcccgg ggtccccagc cgatcgggtc 3181 aggcaagtcc atcccgtcct gagagcccca ggcccccctt cgacaagctt cgaattctgc 3241 agtcgacggt accgcgggcc cgggatccac cggtcgccac catggtgagc aagggcgagg 3301 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca 3361 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt 3421 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct 3481 acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt 3541 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact 3601 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga 3661 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca 3721 acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca 3781 agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca 3841 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg 3901 ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg 3961 ccgccgggat cactctcggc atggacgagc tgtacaagta aagcggccgc gactctagat 4021 cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct 4081 ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc 4141 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc 4201 actgcattct agttgtggtt tgtccaaact catcaatgta tcttaaggcg taaattgtaa 4261 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc 4321 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga 4381 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag 4441 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt 4501 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta 4561 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag 4621 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg 4681 cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa 4741 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac 4801 cctgataaat gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct 4861 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat 4921 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc 4981 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac 5041 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact 5101 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta 5161 gtgaggaggc ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag 5221 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg 5281 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt 5341 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc 5401 tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt 5461 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag 5521 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg 5581 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag 5641 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg 5701 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga 5761 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca 5821 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc 5881 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg 5941 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct 6001 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc 6061 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg 6121 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct 6181 ggagttcttc gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga 6241 aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt 6301 tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc 6361 ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg 6421 tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta 6481 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa 6541 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc 6601 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat 6661 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga 6721 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt 6781 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata 6841 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac 6901 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg 6961 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg 7021 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag 7081 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct 7141 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc 7201 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt 7261 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataacc //