LOCUS       sequence-213736-        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 L330R Q334R L702R
FEATURES             Location/Qualifiers
     source          1..7319
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        62..365
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        366..569
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     520..540
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3260..3269
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3266..3268,3269..3271,3272..3985)
                     /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(3266..3268,3269..3271,3272..3982)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3311..3332)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3572..3591)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3919..3940
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4108..4229
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4145..4164)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4199..4218
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4236..4691)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4373..4394)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4585..4604
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4718..4822
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4824..5181
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4824..5015
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4847..4867)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5032..5167
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5216..6010
                     /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             5216..6010
                     /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(5270..5289)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5880..5899
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6198..6217)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6242..6289
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6618..7206
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7107..7126
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttagttatta atagtaatca attacggggt cattagttca tagcccatat atggagttcc
       61 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
      121 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
      181 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
      241 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
      301 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
      361 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
      421 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
      481 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
      541 tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc cgctagcgct
      601 accggactca gatcctgcca tgggcaaccg cggcatggaa gatctcatcc cgctggtcaa
      661 ccggctgcaa gacgccttct ctgccatcgg ccagaacgcg gacctcgacc tgccgcagat
      721 cgctgtggtg ggcggccaga gcgccggcaa gagctcggtg ctcgagaatt tcgtaggcag
      781 ggacttcttg cctcgaggat ctggcattgt tacccgacgt cccctggtct tgcagctggt
      841 caatgcaacc acagaatatg ccgagttcct gcactgcaag ggaaagaaat tcaccgactt
      901 cgaggaggtg cgccttgaga tcgaggccga gaccgacagg gtcaccggca ccaacaaggg
      961 catctcgccg gtgcctatca acctccgcgt ctactcgccg cacgtgctga acctgaccct
     1021 ggtggacctg cccggaatga ccaaggtccc ggtgggggac caacctcccg acatcgagtt
     1081 ccagatccga gacatgctta tgcagtttgt caccaaggag aactgcctca tcctggccgt
     1141 gtcccccgcc aactctgacc tggccaattc tgacgccctc aaggtcgcca aggaggtgga
     1201 cccccagggc cagcgcacca tcggggtcat caccaagctg gacctgatgg acgagggcac
     1261 agatgcccgt gatgtgctgg agaacaagct gctccccctg cgcagaggct acattggagt
     1321 ggtgaaccgg agccagaagg acattgatgg caagaaggac attaccgccg ccttggctgc
     1381 tgaacgaaag ttcttcctct cccatccatc ttatcgccac ttggctgacc gtatgggcac
     1441 gccctacctg cagaaggtcc tcaatcagca actgacgaac cacatccggg acacactgcc
     1501 ggggctgcgg aacaagctgc agagccagct actgtccatt gagaaggagg tggaggaata
     1561 caagaacttc cgccctgatg acccagctcg caagaccaag gccctgaggc agatggtcag
     1621 gcagtttgcc gtagactttg agaagcgcat tgagggctca ggagatcaga tcgacaccta
     1681 cgaactgtca gggggagccc gcattaaccg aatcttccac gagcgcttcc ctttcgagct
     1741 ggtcaagatg gagtttgatg agaaggaact ccgaagggag atcagctatg ctatcaagaa
     1801 tatccatggc attagaacgg ggctgtttac cccagacatg gcctttgaga ccattgtgaa
     1861 aaagcaggtg aagaagatcc gagaaccgtg tctcaagtgt gtggacatgg ttatctcgga
     1921 gctaatcagc accgttagac agtgcaccaa gaagctccag cagtacccgc ggctacggga
     1981 ggagatggag cgcatcgtga ccacccacat ccgggagcgc gagggccgca ctaaggagca
     2041 ggtcatgctt ctcatcgata tcgagctggc ttacatgaac accaaccatg aggacttcat
     2101 aggctttgcc aatgctcagc agaggagcaa ccagatgaac aagaagaaga cttcagggaa
     2161 ccaggatgag attctggtca tccgcaaggg ctggctgact atcaataata ttggcatcat
     2221 gaaagggggc tccaaggagt actggtttgt gctgactgct gagaatctgt cctggtacaa
     2281 ggatgatgag gagaaagaga agaaatacat gctgtctgtg gacaacctca agctgcggga
     2341 cgtggagaag ggctttatgt cgagcaagca tatctttgcc ctctttaaca cggagcagag
     2401 gaatgtctac aaggattatc ggcagctgga gctagcctgt gagacacagg aggaggtgga
     2461 cagctggaag gcctccttcc tgagggctgg cgtgtaccct gagcgtgttg gggacaaaga
     2521 gaaagccagc gagaccgagg agaatggctc cgacagcttc atgcattcca tggacccaca
     2581 gctggaacgg caagtggaga ccatccggaa tcttgtggac tcatacatgg ccattgtcaa
     2641 caagaccgtg agggacctca tgcccaagac catcatgcac ctcatgatta acaataccaa
     2701 ggagttcatc ttctcggagc tgagggccaa cctgtactcg tgtggggacc agaacacgct
     2761 gatggaggag tcggcggagc aggcacagcg gcgcgacgag atgctgcgca tgtaccacgc
     2821 actgaaggag gcgctcagca tcatcggcaa catcaacacg accaccgtca gcacgcccat
     2881 gcccccgccc gtggacgact cctggctgca ggtgcagagc gtaccggccg gacgcaggtc
     2941 gcccacgtcc agccccacgc cgcagcgccg agcccccgcc gtgcccccag cccggcccgg
     3001 gtcgcggggc cctgctcctg ggcctccgcc tgctgggtcc gccctggggg gggcgccccc
     3061 cgtgccctcc aggccggggg cttcccctga ccctttcggc cctccccctc aggtgccctc
     3121 gcgccccaac cgcgccccgc ccggggtccc cagccgatcg ggtcaggcaa gtccatcccg
     3181 tcctgagagc cccaggcccc ccttcgacaa gcttcgaatt ctgcagtcga cggtaccgcg
     3241 ggcccgggat ccaccggtcg ccaccatggt gagcaagggc gaggagctgt tcaccggggt
     3301 ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg
     3361 cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg
     3421 caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt
     3481 cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg
     3541 ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga
     3601 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa
     3661 ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta
     3721 tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat
     3781 cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg
     3841 ccccgtgctg ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc
     3901 caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct
     3961 cggcatggac gagctgtaca agtaaagcgg ccgcgactct agatcataat cagccatacc
     4021 acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa
     4081 cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa
     4141 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     4201 ggtttgtcca aactcatcaa tgtatcttaa ggcgtaaatt gtaagcgtta atattttgtt
     4261 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     4321 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
     4381 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     4441 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
     4501 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     4561 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
     4621 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     4681 acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     4741 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     4801 ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag
     4861 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     4921 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     4981 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     5041 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     5101 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     5161 ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat
     5221 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     5281 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     5341 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga
     5401 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     5461 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     5521 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     5581 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     5641 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     5701 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga
     5761 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     5821 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     5881 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     5941 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     6001 gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca
     6061 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     6121 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     6181 cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg
     6241 acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg
     6301 gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac
     6361 gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc
     6421 gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag
     6481 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     6541 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     6601 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     6661 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     6721 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     6781 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     6841 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     6901 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     6961 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     7021 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     7081 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     7141 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     7201 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     7261 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgccatgca
//
