LOCUS       sequence-213732-        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 T292A L293A P294A
FEATURES             Location/Qualifiers
     source          1..7319
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        55..358
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        359..562
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     513..533
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3253..3262
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3259..3261,3262..3264,3265..3978)
                     /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(3259..3261,3262..3264,3265..3975)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3304..3325)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3565..3584)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3912..3933
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4101..4222
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4138..4157)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4192..4211
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4229..4684)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4366..4387)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4578..4597
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4711..4815
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4817..5174
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4817..5008
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4840..4860)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5025..5160
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5209..6003
                     /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             5209..6003
                     /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(5263..5282)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5873..5892
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6191..6210)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6235..6282
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6611..7199
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7100..7119
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
       61 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
      121 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
      181 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
      241 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
      301 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
      361 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
      421 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
      481 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
      541 ggaggtctat ataagcagag ctggtttagt gaaccgtcag atccgctagc gctaccggac
      601 tcagatcctg ccatgggcaa ccgcggcatg gaagatctca tcccgctggt caaccggctg
      661 caagacgcct tctctgccat cggccagaac gcggacctcg acctgccgca gatcgctgtg
      721 gtgggcggcc agagcgccgg caagagctcg gtgctcgaga atttcgtagg cagggacttc
      781 ttgcctcgag gatctggcat tgttacccga cgtcccctgg tcttgcagct ggtcaatgca
      841 accacagaat atgccgagtt cctgcactgc aagggaaaga aattcaccga cttcgaggag
      901 gtgcgccttg agatcgaggc cgagaccgac agggtcaccg gcaccaacaa gggcatctcg
      961 ccggtgccta tcaacctccg cgtctactcg ccgcacgtgc tgaacctgac cctggtggac
     1021 ctgcccggaa tgaccaaggt cccggtgggg gaccaacctc ccgacatcga gttccagatc
     1081 cgagacatgc ttatgcagtt tgtcaccaag gagaactgcc tcatcctggc cgtgtccccc
     1141 gccaactctg acctggccaa ttctgacgcc ctcaaggtcg ccaaggaggt ggacccccag
     1201 ggccagcgca ccatcggggt catcaccaag ctggacctga tggacgaggg cacagatgcc
     1261 cgtgatgtgc tggagaacaa gctgctcccc ctgcgcagag gctacattgg agtggtgaac
     1321 cggagccaga aggacattga tggcaagaag gacattaccg ccgccttggc tgctgaacga
     1381 aagttcttcc tctcccatcc atcttatcgc cacttggctg accgtatggg cacgccctac
     1441 ctgcagaagg tcctcaatca gcaactgacg aaccacatcc gggacgccgc cgccgggctg
     1501 cggaacaagc tgcagagcca gctactgtcc attgagaagg aggtggagga atacaagaac
     1561 ttccgccctg atgacccagc tcgcaagacc aaggccctgc tgcagatggt ccagcagttt
     1621 gccgtagact ttgagaagcg cattgagggc tcaggagatc agatcgacac ctacgaactg
     1681 tcagggggag cccgcattaa ccgaatcttc cacgagcgct tccctttcga gctggtcaag
     1741 atggagtttg atgagaagga actccgaagg gagatcagct atgctatcaa gaatatccat
     1801 ggcattagaa cggggctgtt taccccagac atggcctttg agaccattgt gaaaaagcag
     1861 gtgaagaaga tccgagaacc gtgtctcaag tgtgtggaca tggttatctc ggagctaatc
     1921 agcaccgtta gacagtgcac caagaagctc cagcagtacc cgcggctacg ggaggagatg
     1981 gagcgcatcg tgaccaccca catccgggag cgcgagggcc gcactaagga gcaggtcatg
     2041 cttctcatcg atatcgagct ggcttacatg aacaccaacc atgaggactt cataggcttt
     2101 gccaatgctc agcagaggag caaccagatg aacaagaaga agacttcagg gaaccaggat
     2161 gagattctgg tcatccgcaa gggctggctg actatcaata atattggcat catgaaaggg
     2221 ggctccaagg agtactggtt tgtgctgact gctgagaatc tgtcctggta caaggatgat
     2281 gaggagaaag agaagaaata catgctgtct gtggacaacc tcaagctgcg ggacgtggag
     2341 aagggcttta tgtcgagcaa gcatatcttt gccctcttta acacggagca gaggaatgtc
     2401 tacaaggatt atcggcagct ggagctagcc tgtgagacac aggaggaggt ggacagctgg
     2461 aaggcctcct tcctgagggc tggcgtgtac cctgagcgtg ttggggacaa agagaaagcc
     2521 agcgagaccg aggagaatgg ctccgacagc ttcatgcatt ccatggaccc acagctggaa
     2581 cggcaagtgg agaccatccg gaatcttgtg gactcataca tggccattgt caacaagacc
     2641 gtgagggacc tcatgcccaa gaccatcatg cacctcatga ttaacaatac caaggagttc
     2701 atcttctcgg agctgctggc caacctgtac tcgtgtgggg accagaacac gctgatggag
     2761 gagtcggcgg agcaggcaca gcggcgcgac gagatgctgc gcatgtacca cgcactgaag
     2821 gaggcgctca gcatcatcgg caacatcaac acgaccaccg tcagcacgcc catgcccccg
     2881 cccgtggacg actcctggct gcaggtgcag agcgtaccgg ccggacgcag gtcgcccacg
     2941 tccagcccca cgccgcagcg ccgagccccc gccgtgcccc cagcccggcc cgggtcgcgg
     3001 ggccctgctc ctgggcctcc gcctgctggg tccgccctgg ggggggcgcc ccccgtgccc
     3061 tccaggccgg gggcttcccc tgaccctttc ggccctcccc ctcaggtgcc ctcgcgcccc
     3121 aaccgcgccc cgcccggggt ccccagccga tcgggtcagg caagtccatc ccgtcctgag
     3181 agccccaggc cccccttcga caagcttcga attctgcagt cgacggtacc gcgggcccgg
     3241 gatccaccgg tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
     3301 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc
     3361 gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
     3421 cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc
     3481 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
     3541 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag
     3601 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
     3661 ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg
     3721 gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac
     3781 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
     3841 ctgctgcccg acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag
     3901 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg
     3961 gacgagctgt acaagtaaag cggccgcgac tctagatcat aatcagccat accacatttg
     4021 tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa
     4081 tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca
     4141 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     4201 ccaaactcat caatgtatct taaggcgtaa attgtaagcg ttaatatttt gttaaaattc
     4261 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
     4321 ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag
     4381 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
     4441 gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa
     4501 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
     4561 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
     4621 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
     4681 gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     4741 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     4801 tgaaaaagga agagtcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt
     4861 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     4921 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     4981 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     5041 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     5101 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     5161 taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     5221 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     5281 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     5341 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca
     5401 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     5461 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     5521 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     5581 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     5641 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     5701 ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc
     5761 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     5821 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     5881 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     5941 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     6001 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
     6061 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     6121 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg
     6181 ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa
     6241 taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt
     6301 cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg
     6361 tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca
     6421 acgtcggggc ggcaggccct gccatagcct caggttactc atatatactt tagattgatt
     6481 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     6541 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     6601 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     6661 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     6721 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     6781 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     6841 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     6901 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     6961 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     7021 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     7081 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     7141 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     7201 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     7261 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccatg cattagtta
//
