LOCUS       sequence-213763-        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
FEATURES             Location/Qualifiers
     source          1..7319
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        49..352
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        353..556
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     507..527
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3247..3256
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3253..3255,3256..3258,3259..3972)
                     /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(3253..3255,3256..3258,3259..3969)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3298..3319)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3559..3578)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3906..3927
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4095..4216
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4132..4151)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4186..4205
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4223..4678)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4360..4381)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4572..4591
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4705..4809
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4811..5168
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4811..5002
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4834..4854)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5019..5154
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5203..5997
                     /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             5203..5997
                     /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(5257..5276)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5867..5886
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6185..6204)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6229..6276
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6605..7193
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7094..7113
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
       61 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
      121 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
      181 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
      241 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
      301 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg
      361 gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
      421 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
      481 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt
      541 ctatataagc agagctggtt tagtgaaccg tcagatccgc tagcgctacc ggactcagat
      601 cctgccatgg gcaaccgcgg catggaagat ctcatcccgc tggtcaaccg gctgcaagac
      661 gccttctctg ccatcggcca gaacgcggac ctcgacctgc cgcagatcgc tgtggtgggc
      721 ggccagagcg ccggcaagag ctcggtgctc gagaatttcg taggcaggga cttcttgcct
      781 cgaggatctg gcattgttac ccgacgtccc ctggtcttgc agctggtcaa tgcaaccaca
      841 gaatatgccg agttcctgca ctgcaaggga aagaaattca ccgacttcga ggaggtgcgc
      901 cttgagatcg aggccgagac cgacagggtc accggcacca acaagggcat ctcgccggtg
      961 cctatcaacc tccgcgtcta ctcgccgcac gtgctgaacc tgaccctggt ggacctgccc
     1021 ggaatgacca aggtcccggt gggggaccaa cctcccgaca tcgagttcca gatccgagac
     1081 atgcttatgc agtttgtcac caaggagaac tgcctcatcc tggccgtgtc ccccgccaac
     1141 tctgacctgg ccaattctga cgccctcaag gtcgccaagg aggtggaccc ccagggccag
     1201 cgcaccatcg gggtcatcac caagctggac ctgatggacg agggcacaga tgcccgtgat
     1261 gtgctggaga acaagctgct ccccctgcgc agaggctaca ttggagtggt gaaccggagc
     1321 cagaaggaca ttgatggcaa gaaggacatt accgccgcct tggctgctga acgaaagttc
     1381 ttcctctccc atccatctta tcgccacttg gctgaccgta tgggcacgcc ctacctgcag
     1441 aaggtcctca atcagcaact gacgaaccac atccgggaca cactgccggg gctgcggaac
     1501 aagctgcaga gccagctact gtccattgag aaggaggtgg aggaatacaa gaacttccgc
     1561 cctgatgacc cagctcgcaa gaccaaggcc ctgctgcaga tggtccagca gtttgccgta
     1621 gactttgaga agcgcattga gggctcagga gatcagatcg acacctacga actgtcaggg
     1681 ggagcccgca ttaaccgaat cttccacgag cgcttccctt tcgagctggt caagatggag
     1741 tttgatgaga aggaactccg aagggagatc agctatgcta tcaagaatat ccatggcatt
     1801 agaacggggc tgtttacccc aaggagggcc tttgagacca ttgtgaaaaa gcaggtgaag
     1861 aagatccgag aaccgtgtct caagtgtgtg gacatggtta tctcggagct aatcagcacc
     1921 gttagacagt gcaccaagaa gctccagcag tacccgcggc tacgggagga gatggagcgc
     1981 atcgtgacca cccacatccg ggagcgcgag ggccgcacta aggagcaggt catgcttctc
     2041 atcgatatcg agctggctta catgaacacc aaccatgagg acttcatagg ctttgccaat
     2101 gctcagcaga ggagcaacca gatgaacaag aagaagactt cagggaacca ggatgagatt
     2161 ctggtcatcc gcaagggctg gctgactatc aataatattg gcatcatgaa agggggctcc
     2221 aaggagtact ggtttgtgct gactgctgag aatctgtcct ggtacaagga tgatgaggag
     2281 aaagagaaga aatacatgct gtctgtggac aacctcaagc tgcgggacgt ggagaagggc
     2341 tttatgtcga gcaagcatat ctttgccctc tttaacacgg agcagaggaa tgtctacaag
     2401 gattatcggc agctggagct agcctgtgag acacaggagg aggtggacag ctggaaggcc
     2461 tccttcctga gggctggcgt gtaccctgag cgtgttgggg acaaagagaa agccagcgag
     2521 accgaggaga atggctccga cagcttcatg cattccatgg acccacagct ggaacggcaa
     2581 gtggagacca tccggaatct tgtggactca tacatggcca ttgtcaacaa gaccgtgagg
     2641 gacctcatgc ccaagaccat catgcacctc atgattaaca ataccaagga gttcatcttc
     2701 tcggagctgc tggccaacct gtactcgtgt ggggaccaga acacgctgat ggaggagtcg
     2761 gcggagcagg cacagcggcg cgacgagatg ctgcgcatgt accacgcact gaaggaggcg
     2821 ctcagcatca tcggcaacat caacacgacc accgtcagca cgcccatgcc cccgcccgtg
     2881 gacgactcct ggctgcaggt gcagagcgta ccggccggac gcaggtcgcc cacgtccagc
     2941 cccacgccgc agcgccgagc ccccgccgtg cccccagccc ggcccgggtc gcggggccct
     3001 gctcctgggc ctccgcctgc tgggtccgcc ctgggggggg cgccccccgt gccctccagg
     3061 ccgggggctt cccctgaccc tttcggccct ccccctcagg tgccctcgcg ccccaaccgc
     3121 gccccgcccg gggtccccag ccgatcgggt caggcaagtc catcccgtcc tgagagcccc
     3181 aggcccccct tcgacaagct tcgaattctg cagtcgacgg taccgcgggc ccgggatcca
     3241 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
     3301 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
     3361 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
     3421 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
     3481 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
     3541 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
     3601 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     3661 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     3721 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     3781 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     3841 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     3901 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     3961 ctgtacaagt aaagcggccg cgactctaga tcataatcag ccataccaca tttgtagagg
     4021 ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg
     4081 caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     4141 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     4201 tcatcaatgt atcttaaggc gtaaattgta agcgttaata ttttgttaaa attcgcgtta
     4261 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     4321 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     4381 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     4441 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
     4501 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     4561 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
     4621 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca
     4681 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     4741 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     4801 aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa
     4861 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     4921 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     4981 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     5041 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     5101 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     5161 tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     5221 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     5281 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     5341 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg
     5401 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     5461 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     5521 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     5581 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     5641 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     5701 ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg
     5761 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     5821 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     5881 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     5941 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     6001 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     6061 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     6121 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc
     6181 taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa
     6241 gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg
     6301 gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt
     6361 ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg
     6421 gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac
     6481 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     6541 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     6601 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     6661 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     6721 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     6781 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     6841 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     6901 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     6961 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     7021 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     7081 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     7141 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     7201 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     7261 ctgcgttatc ccctgattct gtggataacc gtattaccgc catgcattag ttattaata
//
