LOCUS       sequence-232532-        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 wt
FEATURES             Location/Qualifiers
     source          1..7319
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        80..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        384..587
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     538..558
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3278..3287
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3284..3286,3287..3289,3290..4003)
                     /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(3284..3286,3287..3289,3290..4000)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3329..3350)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3590..3609)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3937..3958
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4126..4247
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4163..4182)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4217..4236
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4254..4709)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4391..4412)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4603..4622
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4736..4840
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4842..5199
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4842..5033
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4865..4885)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5050..5185
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5234..6028
                     /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             5234..6028
                     /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(5288..5307)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5898..5917
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6216..6235)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6260..6307
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6636..7224
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7125..7144
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccgtattacc gccatgcatt agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat
      421 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt
      481 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc
      541 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctggt ttagtgaacc
      601 gtcagatccg ctagcgctac cggactcaga tcctgccatg ggcaaccgcg gcatggaaga
      661 tctcatcccg ctggtcaacc ggctgcaaga cgccttctct gccatcggcc agaacgcgga
      721 cctcgacctg ccgcagatcg ctgtggtggg cggccagagc gccggcaaga gctcggtgct
      781 cgagaatttc gtaggcaggg acttcttgcc tcgaggatct ggcattgtta cccgacgtcc
      841 cctggtcttg cagctggtca atgcaaccac agaatatgcc gagttcctgc actgcaaggg
      901 aaagaaattc accgacttcg aggaggtgcg ccttgagatc gaggccgaga ccgacagggt
      961 caccggcacc aacaagggca tctcgccggt gcctatcaac ctccgcgtct actcgccgca
     1021 cgtgctgaac ctgaccctgg tggacctgcc cggaatgacc aaggtcccgg tgggggacca
     1081 acctcccgac atcgagttcc agatccgaga catgcttatg cagtttgtca ccaaggagaa
     1141 ctgcctcatc ctggccgtgt cccccgccaa ctctgacctg gccaattctg acgccctcaa
     1201 ggtcgccaag gaggtggacc cccagggcca gcgcaccatc ggggtcatca ccaagctgga
     1261 cctgatggac gagggcacag atgcccgtga tgtgctggag aacaagctgc tccccctgcg
     1321 cagaggctac attggagtgg tgaaccggag ccagaaggac attgatggca agaaggacat
     1381 taccgccgcc ttggctgctg aacgaaagtt cttcctctcc catccatctt atcgccactt
     1441 ggctgaccgt atgggcacgc cctacctgca gaaggtcctc aatcagcaac tgacgaacca
     1501 catccgggac acactgccgg ggctgcggaa caagctgcag agccagctac tgtccattga
     1561 gaaggaggtg gaggaataca agaacttccg ccctgatgac ccagctcgca agaccaaggc
     1621 cctgctgcag atggtccagc agtttgccgt agactttgag aagcgcattg agggctcagg
     1681 agatcagatc gacacctacg aactgtcagg gggagcccgc attaaccgaa tcttccacga
     1741 gcgcttccct ttcgagctgg tcaagatgga gtttgatgag aaggaactcc gaagggagat
     1801 cagctatgct atcaagaata tccatggcat tagaacgggg ctgtttaccc cagacatggc
     1861 ctttgagacc attgtgaaaa agcaggtgaa gaagatccga gaaccgtgtc tcaagtgtgt
     1921 ggacatggtt atctcggagc taatcagcac cgttagacag tgcaccaaga agctccagca
     1981 gtacccgcgg ctacgggagg agatggagcg catcgtgacc acccacatcc gggagcgcga
     2041 gggccgcact aaggagcagg tcatgcttct catcgatatc gagctggctt acatgaacac
     2101 caaccatgag gacttcatag gctttgccaa tgctcagcag aggagcaacc agatgaacaa
     2161 gaagaagact tcagggaacc aggatgagat tctggtcatc cgcaagggct ggctgactat
     2221 caataatatt ggcatcatga aagggggctc caaggagtac tggtttgtgc tgactgctga
     2281 gaatctgtcc tggtacaagg atgatgagga gaaagagaag aaatacatgc tgtctgtgga
     2341 caacctcaag ctgcgggacg tggagaaggg ctttatgtcg agcaagcata tctttgccct
     2401 ctttaacacg gagcagagga atgtctacaa ggattatcgg cagctggagc tagcctgtga
     2461 gacacaggag gaggtggaca gctggaaggc ctccttcctg agggctggcg tgtaccctga
     2521 gcgtgttggg gacaaagaga aagccagcga gaccgaggag aatggctccg acagcttcat
     2581 gcattccatg gacccacagc tggaacggca agtggagacc atccggaatc ttgtggactc
     2641 atacatggcc attgtcaaca agaccgtgag ggacctcatg cccaagacca tcatgcacct
     2701 catgattaac aataccaagg agttcatctt ctcggagctg ctggccaacc tgtactcgtg
     2761 tggggaccag aacacgctga tggaggagtc ggcggagcag gcacagcggc gcgacgagat
     2821 gctgcgcatg taccacgcac tgaaggaggc gctcagcatc atcggcaaca tcaacacgac
     2881 caccgtcagc acgcccatgc ccccgcccgt ggacgactcc tggctgcagg tgcagagcgt
     2941 accggccgga cgcaggtcgc ccacgtccag ccccacgccg cagcgccgag cccccgccgt
     3001 gcccccagcc cggcccgggt cgcggggccc tgctcctggg cctccgcctg ctgggtccgc
     3061 cctggggggg gcgccccccg tgccctccag gccgggggct tcccctgacc ctttcggccc
     3121 tccccctcag gtgccctcgc gccccaaccg cgccccgccc ggggtcccca gccgatcggg
     3181 tcaggcaagt ccatcccgtc ctgagagccc caggcccccc ttcgacaagc ttcgaattct
     3241 gcagtcgacg gtaccgcggg cccgggatcc accggtcgcc accatggtga gcaagggcga
     3301 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
     3361 caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa
     3421 gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac
     3481 ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa
     3541 gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa
     3601 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
     3661 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta
     3721 caacagccac aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt
     3781 caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa
     3841 cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc
     3901 cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac
     3961 cgccgccggg atcactctcg gcatggacga gctgtacaag taaagcggcc gcgactctag
     4021 atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac
     4081 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca
     4141 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     4201 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg cgtaaattgt
     4261 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     4321 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     4381 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     4441 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     4501 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     4561 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     4621 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     4681 cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg
     4741 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     4801 accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag
     4861 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     4921 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     4981 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     5041 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     5101 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     5161 tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg
     5221 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     5281 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     5341 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     5401 cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     5461 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     5521 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     5581 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     5641 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     5701 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     5761 gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     5821 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     5881 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     5941 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     6001 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
     6061 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     6121 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     6181 ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg
     6241 gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg
     6301 tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga
     6361 ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg
     6421 ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt
     6481 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     6541 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     6601 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     6661 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     6721 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     6781 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     6841 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     6901 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     6961 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     7021 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     7081 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     7141 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     7201 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     7261 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataa
//
