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 53..356 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 357..560 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 511..531 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 3251..3260 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 3257..3976 /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 3257..3973 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(3302..3323) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(3563..3582) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 3910..3931 /label=EGFP-C /note="EGFP, forward primer" polyA_signal 4099..4220 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4136..4155) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4190..4209 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(4227..4682) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4364..4385) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 4576..4595 /label=F1ori-R /note="F1 origin, reverse primer" promoter 4709..4813 /gene="bla" /label=AmpR promoter promoter 4815..5172 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(4838..4858) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 5023..5158 /label=SV40 ori /note="SV40 origin of replication" primer_bind 5085..5104 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 5207..6001 /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(5261..5280) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 5871..5890 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(6189..6208) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 6233..6280 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 6609..7197 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 7098..7117 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat 61 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa 121 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg 181 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc 241 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct 301 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga 361 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa 421 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc 481 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg 541 aggtctatat aagcagagct ggtttagtga accgtcagat ccgctagcgc taccggactc 601 agatcctgcc atgggcaacc gcggcatgga agatctcatc ccgctggtca accggctgca 661 agacgccttc tctgccatcg gccagaacgc ggacctcgac ctgccgcaga tcgctgtggt 721 gggcggccag agcgccggca agagctcggt gctcgagaat ttcgtaggca gggacttctt 781 gcctcgagga tctggcattg ttacccgacg tcccctggtc ttgcagctgg tcaatgcaac 841 cacagaatat gccgagttcc tgcactgcaa gggaaagaaa ttcaccgact tcgaggaggt 901 gcgccttgag atcgaggccg agaccgacag ggtcaccggc accaacaagg gcatctcgcc 961 ggtgcctatc aacctccgcg tctactcgcc gcacgtgctg aacctgaccc tggtggacct 1021 gcccggaatg accaaggtcc cggtggggga ccaacctccc gacatcgagt tccagatccg 1081 agacatgctt atgcagtttg tcaccaagga gaactgcctc atcctggccg tgtcccccgc 1141 caactctgac ctggccaatt ctgacgccct caaggtcgcc aaggaggtgg acccccaggg 1201 ccagcgcacc atcggggtca tcaccaagct ggacctgatg gacgagggca cagatgcccg 1261 tgatgtgctg gagaacaagc tgctccccct gcgcagaggc tacattggag tggtgaaccg 1321 gagccagaag gacattgatg gcaagaagga cattaccgcc gccttggctg ctgaacgaaa 1381 gttcttcctc tcccatccat cttatcgcca cttggctgac cgtatgggca cgccctacct 1441 gcagaaggtc ctcaatcagc aactgacgaa ccacatcgcc gccgccctgc cggggctgcg 1501 gaacaagctg cagagccagc tactgtccat tgagaaggag gtggaggaat acaagaactt 1561 ccgccctgat gacccagctc gcaagaccaa ggccctgctg cagatggtcc agcagtttgc 1621 cgtagacttt gagaagcgca ttgagggctc aggagatcag atcgacacct acgaactgtc 1681 agggggagcc cgcattaacc gaatcttcca cgagcgcttc cctttcgagc tggtcaagat 1741 ggagtttgat gagaaggaac tccgaaggga gatcagctat gctatcaaga atatccatgg 1801 cattagaacg gggctgttta ccccagacat ggcctttgag accattgtga aaaagcaggt 1861 gaagaagatc cgagaaccgt gtctcaagtg tgtggacatg gttatctcgg agctaatcag 1921 caccgttaga cagtgcacca agaagctcca gcagtacccg cggctacggg aggagatgga 1981 gcgcatcgtg accacccaca tccgggagcg cgagggccgc actaaggagc aggtcatgct 2041 tctcatcgat atcgagctgg cttacatgaa caccaaccat gaggacttca taggctttgc 2101 caatgctcag cagaggagca accagatgaa caagaagaag acttcaggga accaggatga 2161 gattctggtc atccgcaagg gctggctgac tatcaataat attggcatca tgaaaggggg 2221 ctccaaggag tactggtttg tgctgactgc tgagaatctg tcctggtaca aggatgatga 2281 ggagaaagag aagaaataca tgctgtctgt ggacaacctc aagctgcggg acgtggagaa 2341 gggctttatg tcgagcaagc atatctttgc cctctttaac acggagcaga ggaatgtcta 2401 caaggattat cggcagctgg agctagcctg tgagacacag gaggaggtgg acagctggaa 2461 ggcctccttc ctgagggctg gcgtgtaccc tgagcgtgtt ggggacaaag agaaagccag 2521 cgagaccgag gagaatggct ccgacagctt catgcattcc atggacccac agctggaacg 2581 gcaagtggag accatccgga atcttgtgga ctcatacatg gccattgtca acaagaccgt 2641 gagggacctc atgcccaaga ccatcatgca cctcatgatt aacaatacca aggagttcat 2701 cttctcggag ctgctggcca acctgtactc gtgtggggac cagaacacgc tgatggagga 2761 gtcggcggag caggcacagc ggcgcgacga gatgctgcgc atgtaccacg cactgaagga 2821 ggcgctcagc atcatcggca acatcaacac gaccaccgtc agcacgccca tgcccccgcc 2881 cgtggacgac tcctggctgc aggtgcagag cgtaccggcc ggacgcaggt cgcccacgtc 2941 cagccccacg ccgcagcgcc gagcccccgc cgtgccccca gcccggcccg ggtcgcgggg 3001 ccctgctcct gggcctccgc ctgctgggtc cgccctgggg ggggcgcccc ccgtgccctc 3061 caggccgggg gcttcccctg accctttcgg ccctccccct caggtgccct cgcgccccaa 3121 ccgcgccccg cccggggtcc ccagccgatc gggtcaggca agtccatccc gtcctgagag 3181 ccccaggccc cccttcgaca agcttcgaat tctgcagtcg acggtaccgc gggcccggga 3241 tccaccggtc gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat 3301 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga 3361 gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc 3421 cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta 3481 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca 3541 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt 3601 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg 3661 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc 3721 cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg 3781 cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct 3841 gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa 3901 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga 3961 cgagctgtac aagtaaagcg gccgcgactc tagatcataa tcagccatac cacatttgta 4021 gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg 4081 aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat 4141 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc 4201 aaactcatca atgtatctta aggcgtaaat tgtaagcgtt aatattttgt taaaattcgc 4261 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc 4321 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag 4381 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga 4441 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc 4501 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa 4561 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt 4621 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcaccgc tacagggcgc 4681 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat 4741 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg 4801 aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt 4861 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca 4921 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat 4981 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg 5041 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc 5101 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta 5161 ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga 5221 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc 5281 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc 5341 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc 5401 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac 5461 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc 5521 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac 5581 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg 5641 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct 5701 cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt 5761 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg 5821 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac 5881 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg 5941 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg 6001 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat 6061 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc 6121 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg 6181 aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata 6241 aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc 6301 cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt 6361 tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac 6421 gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta gattgattta 6481 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc 6541 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa 6601 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca 6661 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta 6721 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc 6781 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca 6841 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta 6901 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag 6961 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt 7021 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc 7081 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac 7141 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac 7201 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc 7261 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgccatgca ttagttatt //