LOCUS       sequence-213764-        7319 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid of GFP-tagged Dynamin protein..
ACCESSION   .
VERSION     .
KEYWORDS    .
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 Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/s41586-018-0378-6"; journalName: "Nature"; date: "2018-08";
            volume: "560"; issue: "7717"; pages: "258-262"
COMMENT     Sequence Label: pEGFP-N1 Dynamin1 D406R M407R F409A
FEATURES             Location/Qualifiers
     source          1..7319
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        57..360
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        361..564
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     515..535
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3255..3264
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3261..3263,3264..3266,3267..3980)
                     /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             join(3261..3263,3264..3266,3267..3977)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3306..3327)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3567..3586)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3914..3935
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4103..4224
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4140..4159)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4194..4213
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4231..4686)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4368..4389)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4580..4599
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4713..4817
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4819..5176
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4819..5010
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4842..4862)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5027..5162
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5211..6005
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             5211..6005
                     /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(5265..5284)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5875..5894
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6193..6212)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6237..6284
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6613..7201
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7102..7121
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
       61 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
      121 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
      181 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
      241 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
      301 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
      361 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
      421 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      481 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      541 tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta gcgctaccgg
      601 actcagatcc tgccatgggc aaccgcggca tggaagatct catcccgctg gtcaaccggc
      661 tgcaagacgc cttctctgcc atcggccaga acgcggacct cgacctgccg cagatcgctg
      721 tggtgggcgg ccagagcgcc ggcaagagct cggtgctcga gaatttcgta ggcagggact
      781 tcttgcctcg aggatctggc attgttaccc gacgtcccct ggtcttgcag ctggtcaatg
      841 caaccacaga atatgccgag ttcctgcact gcaagggaaa gaaattcacc gacttcgagg
      901 aggtgcgcct tgagatcgag gccgagaccg acagggtcac cggcaccaac aagggcatct
      961 cgccggtgcc tatcaacctc cgcgtctact cgccgcacgt gctgaacctg accctggtgg
     1021 acctgcccgg aatgaccaag gtcccggtgg gggaccaacc tcccgacatc gagttccaga
     1081 tccgagacat gcttatgcag tttgtcacca aggagaactg cctcatcctg gccgtgtccc
     1141 ccgccaactc tgacctggcc aattctgacg ccctcaaggt cgccaaggag gtggaccccc
     1201 agggccagcg caccatcggg gtcatcacca agctggacct gatggacgag ggcacagatg
     1261 cccgtgatgt gctggagaac aagctgctcc ccctgcgcag aggctacatt ggagtggtga
     1321 accggagcca gaaggacatt gatggcaaga aggacattac cgccgccttg gctgctgaac
     1381 gaaagttctt cctctcccat ccatcttatc gccacttggc tgaccgtatg ggcacgccct
     1441 acctgcagaa ggtcctcaat cagcaactga cgaaccacat ccgggacaca ctgccggggc
     1501 tgcggaacaa gctgcagagc cagctactgt ccattgagaa ggaggtggag gaatacaaga
     1561 acttccgccc tgatgaccca gctcgcaaga ccaaggccct gctgcagatg gtccagcagt
     1621 ttgccgtaga ctttgagaag cgcattgagg gctcaggaga tcagatcgac acctacgaac
     1681 tgtcaggggg agcccgcatt aaccgaatct tccacgagcg cttccctttc gagctggtca
     1741 agatggagtt tgatgagaag gaactccgaa gggagatcag ctatgctatc aagaatatcc
     1801 atggcattag aacggggctg tttaccccaa ggagggccgc cgagaccatt gtgaaaaagc
     1861 aggtgaagaa gatccgagaa ccgtgtctca agtgtgtgga catggttatc tcggagctaa
     1921 tcagcaccgt tagacagtgc accaagaagc tccagcagta cccgcggcta cgggaggaga
     1981 tggagcgcat cgtgaccacc cacatccggg agcgcgaggg ccgcactaag gagcaggtca
     2041 tgcttctcat cgatatcgag ctggcttaca tgaacaccaa ccatgaggac ttcataggct
     2101 ttgccaatgc tcagcagagg agcaaccaga tgaacaagaa gaagacttca gggaaccagg
     2161 atgagattct ggtcatccgc aagggctggc tgactatcaa taatattggc atcatgaaag
     2221 ggggctccaa ggagtactgg tttgtgctga ctgctgagaa tctgtcctgg tacaaggatg
     2281 atgaggagaa agagaagaaa tacatgctgt ctgtggacaa cctcaagctg cgggacgtgg
     2341 agaagggctt tatgtcgagc aagcatatct ttgccctctt taacacggag cagaggaatg
     2401 tctacaagga ttatcggcag ctggagctag cctgtgagac acaggaggag gtggacagct
     2461 ggaaggcctc cttcctgagg gctggcgtgt accctgagcg tgttggggac aaagagaaag
     2521 ccagcgagac cgaggagaat ggctccgaca gcttcatgca ttccatggac ccacagctgg
     2581 aacggcaagt ggagaccatc cggaatcttg tggactcata catggccatt gtcaacaaga
     2641 ccgtgaggga cctcatgccc aagaccatca tgcacctcat gattaacaat accaaggagt
     2701 tcatcttctc ggagctgctg gccaacctgt actcgtgtgg ggaccagaac acgctgatgg
     2761 aggagtcggc ggagcaggca cagcggcgcg acgagatgct gcgcatgtac cacgcactga
     2821 aggaggcgct cagcatcatc ggcaacatca acacgaccac cgtcagcacg cccatgcccc
     2881 cgcccgtgga cgactcctgg ctgcaggtgc agagcgtacc ggccggacgc aggtcgccca
     2941 cgtccagccc cacgccgcag cgccgagccc ccgccgtgcc cccagcccgg cccgggtcgc
     3001 ggggccctgc tcctgggcct ccgcctgctg ggtccgccct ggggggggcg ccccccgtgc
     3061 cctccaggcc gggggcttcc cctgaccctt tcggccctcc ccctcaggtg ccctcgcgcc
     3121 ccaaccgcgc cccgcccggg gtccccagcc gatcgggtca ggcaagtcca tcccgtcctg
     3181 agagccccag gccccccttc gacaagcttc gaattctgca gtcgacggta ccgcgggccc
     3241 gggatccacc ggtcgccacc atggtgagca agggcgagga gctgttcacc ggggtggtgc
     3301 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
     3361 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc
     3421 tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc
     3481 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
     3541 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
     3601 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
     3661 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca
     3721 tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg
     3781 acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
     3841 tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg
     3901 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca
     3961 tggacgagct gtacaagtaa agcggccgcg actctagatc ataatcagcc ataccacatt
     4021 tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa
     4081 aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag
     4141 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     4201 gtccaaactc atcaatgtat cttaaggcgt aaattgtaag cgttaatatt ttgttaaaat
     4261 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     4321 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     4381 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     4441 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     4501 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     4561 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     4621 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     4681 gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     4741 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     4801 attgaaaaag gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg
     4861 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     4921 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     4981 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     5041 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     5101 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     5161 cctaggcttt tgcaaagatc gatcaagaga caggatgagg atcgtttcgc atgattgaac
     5221 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     5281 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     5341 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caagacgagg
     5401 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     5461 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     5521 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     5581 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     5641 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     5701 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc
     5761 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     5821 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     5881 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     5941 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     6001 tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     6061 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     6121 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccctag
     6181 ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc
     6241 aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt gttcataaac gcggggttcg
     6301 gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg
     6361 cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc
     6421 caacgtcggg gcggcaggcc ctgccatagc ctcaggttac tcatatatac tttagattga
     6481 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     6541 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     6601 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     6661 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     6721 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     6781 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     6841 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     6901 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     6961 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     7021 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     7081 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     7141 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     7201 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     7261 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcca tgcattagt
//
