LOCUS       sequence-211029-        7319 bp    DNA     circular SYN 15-APR-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 Apr 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        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3259..3268
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3265..3267,3268..3270,3271..3984)
                     /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(3265..3267,3268..3270,3271..3981)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3310..3331)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3571..3590)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3918..3939
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4107..4228
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4144..4163)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4198..4217
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4235..4690)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4372..4393)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4584..4603
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4717..4821
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4823..5180
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4823..5014
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4846..4866)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5031..5166
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5215..6009
                     /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             5215..6009
                     /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(5269..5288)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5879..5898
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6197..6216)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6241..6288
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6617..7205
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7106..7125
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atcctgccat gggcaaccgc ggcatggaag atctcatccc gctggtcaac
      661 cggctgcaag acgccttctc tgccatcggc cagaacgcgg acctcgacct gccgcagatc
      721 gctgtggtgg gcggccagag cgccggcaag agctcggtgc tcgagaattt cgtaggcagg
      781 gacttcttgc ctcgaggatc tggcattgtt acccgacgtc ccctggtctt gcagctggtc
      841 aatgcaacca cagaatatgc cgagttcctg cactgcaagg gaaagaaatt caccgacttc
      901 gaggaggtgc gccttgagat cgaggccgag accgacaggg tcaccggcac caacaagggc
      961 atctcgccgg tgcctatcaa cctccgcgtc tactcgccgc acgtgctgaa cctgaccctg
     1021 gtggacctgc ccggaatgac caaggtcccg gtgggggacc aacctcccga catcgagttc
     1081 cagatccgag acatgcttat gcagtttgtc accaaggaga actgcctcat cctggccgtg
     1141 tcccccgcca actctgacct ggccaattct gacgccctca aggtcgccaa ggaggtggac
     1201 ccccagggcc agcgcaccat cggggtcatc accaagctgg acctgatgga cgagggcaca
     1261 gatgcccgtg atgtgctgga gaacaagctg ctccccctgc gcagaggcta cattggagtg
     1321 gtgaaccgga gccagaagga cattgatggc aagaaggaca ttaccgccgc cttggctgct
     1381 gaacgaaagt tcttcctctc ccatccatct tatcgccact tggctgaccg tatgggcacg
     1441 ccctacctgc agaaggtcct caatcagcaa ctgacgaacc acatccggga cgccgccgcc
     1501 gggctgcgga acaagctgca gagccagcta ctgtccattg agaaggaggt ggaggaatac
     1561 aagaacttcc gccctgatga cccagctcgc aagaccaagg ccctgctgca gatggtccag
     1621 cagtttgccg tagactttga gaagcgcatt gagggctcag gagatcagat cgacacctac
     1681 gaactgtcag ggggagcccg cattaaccga atcttccacg agcgcttccc tttcgagctg
     1741 gtcaagatgg agtttgatga gaaggaactc cgaagggaga tcagctatgc tatcaagaat
     1801 atccatggca ttagaacggg gctgtttacc ccagacatgg cctttgagac cattgtgaaa
     1861 aagcaggtga agaagatccg agaaccgtgt ctcaagtgtg tggacatggt tatctcggag
     1921 ctaatcagca ccgttagaca gtgcaccaag aagctccagc agtacccgcg gctacgggag
     1981 gagatggagc gcatcgtgac cacccacatc cgggagcgcg agggccgcac taaggagcag
     2041 gtcatgcttc tcatcgatat cgagctggct tacatgaaca ccaaccatga ggacttcata
     2101 ggctttgcca atgctcagca gaggagcaac cagatgaaca agaagaagac ttcagggaac
     2161 caggatgaga ttctggtcat ccgcaagggc tggctgacta tcaataatat tggcatcatg
     2221 aaagggggct ccaaggagta ctggtttgtg ctgactgctg agaatctgtc ctggtacaag
     2281 gatgatgagg agaaagagaa gaaatacatg ctgtctgtgg acaacctcaa gctgcgggac
     2341 gtggagaagg gctttatgtc gagcaagcat atctttgccc tctttaacac ggagcagagg
     2401 aatgtctaca aggattatcg gcagctggag ctagcctgtg agacacagga ggaggtggac
     2461 agctggaagg cctccttcct gagggctggc gtgtaccctg agcgtgttgg ggacaaagag
     2521 aaagccagcg agaccgagga gaatggctcc gacagcttca tgcattccat ggacccacag
     2581 ctggaacggc aagtggagac catccggaat cttgtggact catacatggc cattgtcaac
     2641 aagaccgtga gggacctcat gcccaagacc atcatgcacc tcatgattaa caataccaag
     2701 gagttcatct tctcggagct gctggccaac ctgtactcgt gtggggacca gaacacgctg
     2761 atggaggagt cggcggagca ggcacagcgg cgcgacgaga tgctgcgcat gtaccacgca
     2821 ctgaaggagg cgctcagcat catcggcaac atcaacacga ccaccgtcag cacgcccatg
     2881 cccccgcccg tggacgactc ctggctgcag gtgcagagcg taccggccgg acgcaggtcg
     2941 cccacgtcca gccccacgcc gcagcgccga gcccccgccg tgcccccagc ccggcccggg
     3001 tcgcggggcc ctgctcctgg gcctccgcct gctgggtccg ccctgggggg ggcgcccccc
     3061 gtgccctcca ggccgggggc ttcccctgac cctttcggcc ctccccctca ggtgccctcg
     3121 cgccccaacc gcgccccgcc cggggtcccc agccgatcgg gtcaggcaag tccatcccgt
     3181 cctgagagcc ccaggccccc cttcgacaag cttcgaattc tgcagtcgac ggtaccgcgg
     3241 gcccgggatc caccggtcgc caccatggtg agcaagggcg aggagctgtt caccggggtg
     3301 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
     3361 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
     3421 aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc
     3481 agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
     3541 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
     3601 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     3661 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat
     3721 atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc
     3781 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
     3841 cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc
     3901 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
     3961 ggcatggacg agctgtacaa gtaaagcggc cgcgactcta gatcataatc agccatacca
     4021 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
     4081 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
     4141 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     4201 gtttgtccaa actcatcaat gtatcttaag gcgtaaattg taagcgttaa tattttgtta
     4261 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     4321 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     4381 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     4441 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     4501 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     4561 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     4621 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     4681 cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     4741 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     4801 taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt
     4861 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     4921 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     4981 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     5041 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     5101 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     5161 gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt
     5221 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     5281 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     5341 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac
     5401 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     5461 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     5521 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     5581 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     5641 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     5701 caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag
     5761 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     5821 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     5881 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     5941 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     6001 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     6061 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     6121 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     6181 ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga
     6241 cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg
     6301 ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg
     6361 cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg
     6421 cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga
     6481 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     6541 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     6601 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     6661 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     6721 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
     6781 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     6841 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     6901 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     6961 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     7021 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     7081 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     7141 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     7201 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     7261 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gccatgcat
//
