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 L330R Q334R L702R 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 62..365 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 366..569 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 520..540 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 3260..3269 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 3266..3985 /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 3266..3982 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(3311..3332) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(3572..3591) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 3919..3940 /label=EGFP-C /note="EGFP, forward primer" polyA_signal 4108..4229 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4145..4164) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4199..4218 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(4236..4691) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4373..4394) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 4585..4604 /label=F1ori-R /note="F1 origin, reverse primer" promoter 4718..4822 /gene="bla" /label=AmpR promoter promoter 4824..5181 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(4847..4867) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 5032..5167 /label=SV40 ori /note="SV40 origin of replication" primer_bind 5094..5113 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 5216..6010 /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(5270..5289) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 5880..5899 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(6198..6217) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 6242..6289 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 6618..7206 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 7107..7126 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 ttagttatta atagtaatca attacggggt cattagttca tagcccatat atggagttcc 61 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat 121 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc 181 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc 241 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt 301 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta 361 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg 421 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac 481 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg 541 tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc cgctagcgct 601 accggactca gatcctgcca tgggcaaccg cggcatggaa gatctcatcc cgctggtcaa 661 ccggctgcaa gacgccttct ctgccatcgg ccagaacgcg gacctcgacc tgccgcagat 721 cgctgtggtg ggcggccaga gcgccggcaa gagctcggtg ctcgagaatt tcgtaggcag 781 ggacttcttg cctcgaggat ctggcattgt tacccgacgt cccctggtct tgcagctggt 841 caatgcaacc acagaatatg ccgagttcct gcactgcaag ggaaagaaat tcaccgactt 901 cgaggaggtg cgccttgaga tcgaggccga gaccgacagg gtcaccggca ccaacaaggg 961 catctcgccg gtgcctatca acctccgcgt ctactcgccg cacgtgctga acctgaccct 1021 ggtggacctg cccggaatga ccaaggtccc ggtgggggac caacctcccg acatcgagtt 1081 ccagatccga gacatgctta tgcagtttgt caccaaggag aactgcctca tcctggccgt 1141 gtcccccgcc aactctgacc tggccaattc tgacgccctc aaggtcgcca aggaggtgga 1201 cccccagggc cagcgcacca tcggggtcat caccaagctg gacctgatgg acgagggcac 1261 agatgcccgt gatgtgctgg agaacaagct gctccccctg cgcagaggct acattggagt 1321 ggtgaaccgg agccagaagg acattgatgg caagaaggac attaccgccg ccttggctgc 1381 tgaacgaaag ttcttcctct cccatccatc ttatcgccac ttggctgacc gtatgggcac 1441 gccctacctg cagaaggtcc tcaatcagca actgacgaac cacatccggg acacactgcc 1501 ggggctgcgg aacaagctgc agagccagct actgtccatt gagaaggagg tggaggaata 1561 caagaacttc cgccctgatg acccagctcg caagaccaag gccctgaggc agatggtcag 1621 gcagtttgcc gtagactttg agaagcgcat tgagggctca ggagatcaga tcgacaccta 1681 cgaactgtca gggggagccc gcattaaccg aatcttccac gagcgcttcc ctttcgagct 1741 ggtcaagatg gagtttgatg agaaggaact ccgaagggag atcagctatg ctatcaagaa 1801 tatccatggc attagaacgg ggctgtttac cccagacatg gcctttgaga ccattgtgaa 1861 aaagcaggtg aagaagatcc gagaaccgtg tctcaagtgt gtggacatgg ttatctcgga 1921 gctaatcagc accgttagac agtgcaccaa gaagctccag cagtacccgc ggctacggga 1981 ggagatggag cgcatcgtga ccacccacat ccgggagcgc gagggccgca ctaaggagca 2041 ggtcatgctt ctcatcgata tcgagctggc ttacatgaac accaaccatg aggacttcat 2101 aggctttgcc aatgctcagc agaggagcaa ccagatgaac aagaagaaga cttcagggaa 2161 ccaggatgag attctggtca tccgcaaggg ctggctgact atcaataata ttggcatcat 2221 gaaagggggc tccaaggagt actggtttgt gctgactgct gagaatctgt cctggtacaa 2281 ggatgatgag gagaaagaga agaaatacat gctgtctgtg gacaacctca agctgcggga 2341 cgtggagaag ggctttatgt cgagcaagca tatctttgcc ctctttaaca cggagcagag 2401 gaatgtctac aaggattatc ggcagctgga gctagcctgt gagacacagg aggaggtgga 2461 cagctggaag gcctccttcc tgagggctgg cgtgtaccct gagcgtgttg gggacaaaga 2521 gaaagccagc gagaccgagg agaatggctc cgacagcttc atgcattcca tggacccaca 2581 gctggaacgg caagtggaga ccatccggaa tcttgtggac tcatacatgg ccattgtcaa 2641 caagaccgtg agggacctca tgcccaagac catcatgcac ctcatgatta acaataccaa 2701 ggagttcatc ttctcggagc tgagggccaa cctgtactcg tgtggggacc agaacacgct 2761 gatggaggag tcggcggagc aggcacagcg gcgcgacgag atgctgcgca tgtaccacgc 2821 actgaaggag gcgctcagca tcatcggcaa catcaacacg accaccgtca gcacgcccat 2881 gcccccgccc gtggacgact cctggctgca ggtgcagagc gtaccggccg gacgcaggtc 2941 gcccacgtcc agccccacgc cgcagcgccg agcccccgcc gtgcccccag cccggcccgg 3001 gtcgcggggc cctgctcctg ggcctccgcc tgctgggtcc gccctggggg gggcgccccc 3061 cgtgccctcc aggccggggg cttcccctga ccctttcggc cctccccctc aggtgccctc 3121 gcgccccaac cgcgccccgc ccggggtccc cagccgatcg ggtcaggcaa gtccatcccg 3181 tcctgagagc cccaggcccc ccttcgacaa gcttcgaatt ctgcagtcga cggtaccgcg 3241 ggcccgggat ccaccggtcg ccaccatggt gagcaagggc gaggagctgt tcaccggggt 3301 ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg 3361 cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg 3421 caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt 3481 cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg 3541 ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga 3601 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa 3661 ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta 3721 tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat 3781 cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg 3841 ccccgtgctg ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc 3901 caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct 3961 cggcatggac gagctgtaca agtaaagcgg ccgcgactct agatcataat cagccatacc 4021 acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa 4081 cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa 4141 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt 4201 ggtttgtcca aactcatcaa tgtatcttaa ggcgtaaatt gtaagcgtta atattttgtt 4261 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg 4321 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg 4381 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta 4441 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg 4501 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa 4561 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct 4621 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct 4681 acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt 4741 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca 4801 ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag 4861 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc 4921 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa 4981 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc 5041 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat 5101 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt 5161 ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat 5221 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta 5281 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca 5341 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga 5401 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga 5461 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct 5521 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg 5581 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga 5641 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca 5701 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga 5761 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg 5821 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc 5881 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt 5941 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga 6001 gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca 6061 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc 6121 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac 6181 cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg 6241 acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg 6301 gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac 6361 gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc 6421 gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag 6481 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat 6541 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa 6601 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca 6661 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt 6721 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg 6781 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc 6841 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga 6901 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc 6961 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc 7021 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca 7081 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg 7141 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta 7201 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct 7261 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgccatgca //