LOCUS       sequence-213761-        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 D406A M407A F409A
FEATURES             Location/Qualifiers
     source          1..7319
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        54..357
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        358..561
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     512..532
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3252..3261
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3258..3260,3261..3263,3264..3977)
                     /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(3258..3260,3261..3263,3264..3974)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3303..3324)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3564..3583)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3911..3932
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4100..4221
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4137..4156)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4191..4210
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4228..4683)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4365..4386)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4577..4596
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4710..4814
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4816..5173
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4816..5007
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4839..4859)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5024..5159
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5208..6002
                     /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             5208..6002
                     /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(5262..5281)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5872..5891
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6190..6209)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6234..6281
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6610..7198
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7099..7118
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca
       61 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
      121 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
      181 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
      241 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
      301 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg
      361 atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca
      421 agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt
      481 ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg
      541 gaggtctata taagcagagc tggtttagtg aaccgtcaga tccgctagcg ctaccggact
      601 cagatcctgc catgggcaac cgcggcatgg aagatctcat cccgctggtc aaccggctgc
      661 aagacgcctt ctctgccatc ggccagaacg cggacctcga cctgccgcag atcgctgtgg
      721 tgggcggcca gagcgccggc aagagctcgg tgctcgagaa tttcgtaggc agggacttct
      781 tgcctcgagg atctggcatt gttacccgac gtcccctggt cttgcagctg gtcaatgcaa
      841 ccacagaata tgccgagttc ctgcactgca agggaaagaa attcaccgac ttcgaggagg
      901 tgcgccttga gatcgaggcc gagaccgaca gggtcaccgg caccaacaag ggcatctcgc
      961 cggtgcctat caacctccgc gtctactcgc cgcacgtgct gaacctgacc ctggtggacc
     1021 tgcccggaat gaccaaggtc ccggtggggg accaacctcc cgacatcgag ttccagatcc
     1081 gagacatgct tatgcagttt gtcaccaagg agaactgcct catcctggcc gtgtcccccg
     1141 ccaactctga cctggccaat tctgacgccc tcaaggtcgc caaggaggtg gacccccagg
     1201 gccagcgcac catcggggtc atcaccaagc tggacctgat ggacgagggc acagatgccc
     1261 gtgatgtgct ggagaacaag ctgctccccc tgcgcagagg ctacattgga gtggtgaacc
     1321 ggagccagaa ggacattgat ggcaagaagg acattaccgc cgccttggct gctgaacgaa
     1381 agttcttcct ctcccatcca tcttatcgcc acttggctga ccgtatgggc acgccctacc
     1441 tgcagaaggt cctcaatcag caactgacga accacatccg ggacacactg ccggggctgc
     1501 ggaacaagct gcagagccag ctactgtcca ttgagaagga ggtggaggaa tacaagaact
     1561 tccgccctga tgacccagct cgcaagacca aggccctgct gcagatggtc cagcagtttg
     1621 ccgtagactt tgagaagcgc attgagggct caggagatca gatcgacacc tacgaactgt
     1681 cagggggagc ccgcattaac cgaatcttcc acgagcgctt ccctttcgag ctggtcaaga
     1741 tggagtttga tgagaaggaa ctccgaaggg agatcagcta tgctatcaag aatatccatg
     1801 gcattagaac ggggctgttt accccagccg ccgccgccga gaccattgtg aaaaagcagg
     1861 tgaagaagat ccgagaaccg tgtctcaagt gtgtggacat ggttatctcg gagctaatca
     1921 gcaccgttag acagtgcacc aagaagctcc agcagtaccc gcggctacgg gaggagatgg
     1981 agcgcatcgt gaccacccac atccgggagc gcgagggccg cactaaggag caggtcatgc
     2041 ttctcatcga tatcgagctg gcttacatga acaccaacca tgaggacttc ataggctttg
     2101 ccaatgctca gcagaggagc aaccagatga acaagaagaa gacttcaggg aaccaggatg
     2161 agattctggt catccgcaag ggctggctga ctatcaataa tattggcatc atgaaagggg
     2221 gctccaagga gtactggttt gtgctgactg ctgagaatct gtcctggtac aaggatgatg
     2281 aggagaaaga gaagaaatac atgctgtctg tggacaacct caagctgcgg gacgtggaga
     2341 agggctttat gtcgagcaag catatctttg ccctctttaa cacggagcag aggaatgtct
     2401 acaaggatta tcggcagctg gagctagcct gtgagacaca ggaggaggtg gacagctgga
     2461 aggcctcctt cctgagggct ggcgtgtacc ctgagcgtgt tggggacaaa gagaaagcca
     2521 gcgagaccga ggagaatggc tccgacagct tcatgcattc catggaccca cagctggaac
     2581 ggcaagtgga gaccatccgg aatcttgtgg actcatacat ggccattgtc aacaagaccg
     2641 tgagggacct catgcccaag accatcatgc acctcatgat taacaatacc aaggagttca
     2701 tcttctcgga gctgctggcc aacctgtact cgtgtgggga ccagaacacg ctgatggagg
     2761 agtcggcgga gcaggcacag cggcgcgacg agatgctgcg catgtaccac gcactgaagg
     2821 aggcgctcag catcatcggc aacatcaaca cgaccaccgt cagcacgccc atgcccccgc
     2881 ccgtggacga ctcctggctg caggtgcaga gcgtaccggc cggacgcagg tcgcccacgt
     2941 ccagccccac gccgcagcgc cgagcccccg ccgtgccccc agcccggccc gggtcgcggg
     3001 gccctgctcc tgggcctccg cctgctgggt ccgccctggg gggggcgccc cccgtgccct
     3061 ccaggccggg ggcttcccct gaccctttcg gccctccccc tcaggtgccc tcgcgcccca
     3121 accgcgcccc gcccggggtc cccagccgat cgggtcaggc aagtccatcc cgtcctgaga
     3181 gccccaggcc ccccttcgac aagcttcgaa ttctgcagtc gacggtaccg cgggcccggg
     3241 atccaccggt cgccaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca
     3301 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg
     3361 agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc
     3421 ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct
     3481 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc
     3541 aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt
     3601 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg
     3661 gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg
     3721 ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg
     3781 gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc
     3841 tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga
     3901 agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg
     3961 acgagctgta caagtaaagc ggccgcgact ctagatcata atcagccata ccacatttgt
     4021 agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat
     4081 gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa
     4141 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     4201 caaactcatc aatgtatctt aaggcgtaaa ttgtaagcgt taatattttg ttaaaattcg
     4261 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
     4321 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
     4381 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
     4441 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag
     4501 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
     4561 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg
     4621 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
     4681 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     4741 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     4801 gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg
     4861 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     4921 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     4981 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     5041 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     5101 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     5161 aggcttttgc aaagatcgat caagagacag gatgaggatc gtttcgcatg attgaacaag
     5221 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
     5281 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
     5341 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag
     5401 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
     5461 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     5521 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     5581 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     5641 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     5701 tcgcgccagc cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg
     5761 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     5821 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     5881 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     5941 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     6001 gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
     6061 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     6121 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accctagggg
     6181 gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat
     6241 aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc
     6301 ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt
     6361 ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa
     6421 cgtcggggcg gcaggccctg ccatagcctc aggttactca tatatacttt agattgattt
     6481 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     6541 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     6601 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     6661 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     6721 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     6781 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     6841 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     6901 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     6961 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     7021 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     7081 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     7141 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     7201 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     7261 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgccatgc attagttat
//
