LOCUS       sequence-215592-        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 T78R L84R T92R V118R
FEATURES             Location/Qualifiers
     source          1..7319
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        56..359
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        360..563
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     514..534
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3254..3263
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3260..3262,3263..3265,3266..3979)
                     /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(3260..3262,3263..3265,3266..3976)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3305..3326)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3566..3585)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3913..3934
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4102..4223
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4139..4158)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4193..4212
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4230..4685)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4367..4388)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4579..4598
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4712..4816
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4818..5175
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4818..5009
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4841..4861)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5026..5161
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5210..6004
                     /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             5210..6004
                     /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(5264..5283)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5874..5893
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6192..6211)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6236..6283
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6612..7200
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7101..7120
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 attaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
       61 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
      121 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
      181 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
      241 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
      301 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg
      361 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
      421 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
      481 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
      541 gggaggtcta tataagcaga gctggtttag tgaaccgtca gatccgctag cgctaccgga
      601 ctcagatcct gccatgggca accgcggcat ggaagatctc atcccgctgg tcaaccggct
      661 gcaagacgcc ttctctgcca tcggccagaa cgcggacctc gacctgccgc agatcgctgt
      721 ggtgggcggc cagagcgccg gcaagagctc ggtgctcgag aatttcgtag gcagggactt
      781 cttgcctcga ggatctggca ttgttacccg acgtcccctg gtcttgcagc tggtcaatgc
      841 aaccagggaa tatgccgagt tcaggcactg caagggaaag aaattcaggg acttcgagga
      901 ggtgcgcctt gagatcgagg ccgagaccga cagggtcacc ggcaccaaca agggcatctc
      961 gccgaggcct atcaacctcc gcgtctactc gccgcacgtg ctgaacctga ccctggtgga
     1021 cctgcccgga atgaccaagg tcccggtggg ggaccaacct cccgacatcg agttccagat
     1081 ccgagacatg cttatgcagt ttgtcaccaa ggagaactgc ctcatcctgg ccgtgtcccc
     1141 cgccaactct gacctggcca attctgacgc cctcaaggtc gccaaggagg tggaccccca
     1201 gggccagcgc accatcgggg tcatcaccaa gctggacctg atggacgagg gcacagatgc
     1261 ccgtgatgtg ctggagaaca agctgctccc cctgcgcaga ggctacattg gagtggtgaa
     1321 ccggagccag aaggacattg atggcaagaa ggacattacc gccgccttgg ctgctgaacg
     1381 aaagttcttc ctctcccatc catcttatcg ccacttggct gaccgtatgg gcacgcccta
     1441 cctgcagaag gtcctcaatc agcaactgac gaaccacatc cgggacacac tgccggggct
     1501 gcggaacaag ctgcagagcc agctactgtc cattgagaag gaggtggagg aatacaagaa
     1561 cttccgccct gatgacccag ctcgcaagac caaggccctg ctgcagatgg tccagcagtt
     1621 tgccgtagac tttgagaagc gcattgaggg ctcaggagat cagatcgaca cctacgaact
     1681 gtcaggggga gcccgcatta accgaatctt ccacgagcgc ttccctttcg agctggtcaa
     1741 gatggagttt gatgagaagg aactccgaag ggagatcagc tatgctatca agaatatcca
     1801 tggcattaga acggggctgt ttaccccaga catggccttt gagaccattg tgaaaaagca
     1861 ggtgaagaag atccgagaac cgtgtctcaa gtgtgtggac atggttatct cggagctaat
     1921 cagcaccgtt agacagtgca ccaagaagct ccagcagtac ccgcggctac gggaggagat
     1981 ggagcgcatc gtgaccaccc acatccggga gcgcgagggc cgcactaagg agcaggtcat
     2041 gcttctcatc gatatcgagc tggcttacat gaacaccaac catgaggact tcataggctt
     2101 tgccaatgct cagcagagga gcaaccagat gaacaagaag aagacttcag ggaaccagga
     2161 tgagattctg gtcatccgca agggctggct gactatcaat aatattggca tcatgaaagg
     2221 gggctccaag gagtactggt ttgtgctgac tgctgagaat ctgtcctggt acaaggatga
     2281 tgaggagaaa gagaagaaat acatgctgtc tgtggacaac ctcaagctgc gggacgtgga
     2341 gaagggcttt atgtcgagca agcatatctt tgccctcttt aacacggagc agaggaatgt
     2401 ctacaaggat tatcggcagc tggagctagc ctgtgagaca caggaggagg tggacagctg
     2461 gaaggcctcc ttcctgaggg ctggcgtgta ccctgagcgt gttggggaca aagagaaagc
     2521 cagcgagacc gaggagaatg gctccgacag cttcatgcat tccatggacc cacagctgga
     2581 acggcaagtg gagaccatcc ggaatcttgt ggactcatac atggccattg tcaacaagac
     2641 cgtgagggac ctcatgccca agaccatcat gcacctcatg attaacaata ccaaggagtt
     2701 catcttctcg gagctgctgg ccaacctgta ctcgtgtggg gaccagaaca cgctgatgga
     2761 ggagtcggcg gagcaggcac agcggcgcga cgagatgctg cgcatgtacc acgcactgaa
     2821 ggaggcgctc agcatcatcg gcaacatcaa cacgaccacc gtcagcacgc ccatgccccc
     2881 gcccgtggac gactcctggc tgcaggtgca gagcgtaccg gccggacgca ggtcgcccac
     2941 gtccagcccc acgccgcagc gccgagcccc cgccgtgccc ccagcccggc ccgggtcgcg
     3001 gggccctgct cctgggcctc cgcctgctgg gtccgccctg gggggggcgc cccccgtgcc
     3061 ctccaggccg ggggcttccc ctgacccttt cggccctccc cctcaggtgc cctcgcgccc
     3121 caaccgcgcc ccgcccgggg tccccagccg atcgggtcag gcaagtccat cccgtcctga
     3181 gagccccagg ccccccttcg acaagcttcg aattctgcag tcgacggtac cgcgggcccg
     3241 ggatccaccg gtcgccacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
     3301 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
     3361 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct
     3421 gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg
     3481 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
     3541 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
     3601 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
     3661 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcat
     3721 ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga
     3781 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
     3841 gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga
     3901 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
     3961 ggacgagctg tacaagtaaa gcggccgcga ctctagatca taatcagcca taccacattt
     4021 gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa
     4081 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc
     4141 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     4201 tccaaactca tcaatgtatc ttaaggcgta aattgtaagc gttaatattt tgttaaaatt
     4261 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     4321 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
     4381 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     4441 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
     4501 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     4561 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
     4621 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
     4681 cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     4741 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     4801 ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg
     4861 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     4921 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     4981 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     5041 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     5101 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     5161 ctaggctttt gcaaagatcg atcaagagac aggatgagga tcgtttcgca tgattgaaca
     5221 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
     5281 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
     5341 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc
     5401 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
     5461 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
     5521 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
     5581 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
     5641 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
     5701 gctcgcgcca gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct
     5761 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     5821 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
     5881 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
     5941 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     6001 ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga
     6061 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     6121 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccctagg
     6181 gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca
     6241 ataaaaagac agaataaaac gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg
     6301 tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc
     6361 gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc
     6421 aacgtcgggg cggcaggccc tgccatagcc tcaggttact catatatact ttagattgat
     6481 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     6541 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     6601 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     6661 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     6721 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     6781 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     6841 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     6901 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     6961 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     7021 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     7081 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     7141 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     7201 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     7261 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat gcattagtt
//
