LOCUS       sequence-213724-        7319 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression 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 P294A
FEATURES             Location/Qualifiers
     source          1..7319
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        52..355
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        356..559
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     510..530
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3250..3259
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3256..3258,3259..3261,3262..3975)
                     /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(3256..3258,3259..3261,3262..3972)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3301..3322)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3562..3581)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3909..3930
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4098..4219
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4135..4154)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4189..4208
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4226..4681)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4363..4384)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4575..4594
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4708..4812
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4814..5171
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4814..5005
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4837..4857)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5022..5157
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5206..6000
                     /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             5206..6000
                     /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(5260..5279)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5870..5889
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6188..6207)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6232..6279
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6608..7196
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7097..7116
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
       61 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
      121 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
      181 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
      241 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
      301 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat
      361 gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag
      421 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
      481 aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga
      541 ggtctatata agcagagctg gtttagtgaa ccgtcagatc cgctagcgct accggactca
      601 gatcctgcca tgggcaaccg cggcatggaa gatctcatcc cgctggtcaa ccggctgcaa
      661 gacgccttct ctgccatcgg ccagaacgcg gacctcgacc tgccgcagat cgctgtggtg
      721 ggcggccaga gcgccggcaa gagctcggtg ctcgagaatt tcgtaggcag ggacttcttg
      781 cctcgaggat ctggcattgt tacccgacgt cccctggtct tgcagctggt caatgcaacc
      841 acagaatatg ccgagttcct gcactgcaag ggaaagaaat tcaccgactt cgaggaggtg
      901 cgccttgaga tcgaggccga gaccgacagg gtcaccggca ccaacaaggg catctcgccg
      961 gtgcctatca acctccgcgt ctactcgccg cacgtgctga acctgaccct ggtggacctg
     1021 cccggaatga ccaaggtccc ggtgggggac caacctcccg acatcgagtt ccagatccga
     1081 gacatgctta tgcagtttgt caccaaggag aactgcctca tcctggccgt gtcccccgcc
     1141 aactctgacc tggccaattc tgacgccctc aaggtcgcca aggaggtgga cccccagggc
     1201 cagcgcacca tcggggtcat caccaagctg gacctgatgg acgagggcac agatgcccgt
     1261 gatgtgctgg agaacaagct gctccccctg cgcagaggct acattggagt ggtgaaccgg
     1321 agccagaagg acattgatgg caagaaggac attaccgccg ccttggctgc tgaacgaaag
     1381 ttcttcctct cccatccatc ttatcgccac ttggctgacc gtatgggcac gccctacctg
     1441 cagaaggtcc tcaatcagca actgacgaac cacatccggg acacactggc cgggctgcgg
     1501 aacaagctgc agagccagct actgtccatt gagaaggagg tggaggaata caagaacttc
     1561 cgccctgatg acccagctcg caagaccaag gccctgctgc agatggtcca gcagtttgcc
     1621 gtagactttg agaagcgcat tgagggctca ggagatcaga tcgacaccta cgaactgtca
     1681 gggggagccc gcattaaccg aatcttccac gagcgcttcc ctttcgagct ggtcaagatg
     1741 gagtttgatg agaaggaact ccgaagggag atcagctatg ctatcaagaa tatccatggc
     1801 attagaacgg ggctgtttac cccagacatg gcctttgaga ccattgtgaa aaagcaggtg
     1861 aagaagatcc gagaaccgtg tctcaagtgt gtggacatgg ttatctcgga gctaatcagc
     1921 accgttagac agtgcaccaa gaagctccag cagtacccgc ggctacggga ggagatggag
     1981 cgcatcgtga ccacccacat ccgggagcgc gagggccgca ctaaggagca ggtcatgctt
     2041 ctcatcgata tcgagctggc ttacatgaac accaaccatg aggacttcat aggctttgcc
     2101 aatgctcagc agaggagcaa ccagatgaac aagaagaaga cttcagggaa ccaggatgag
     2161 attctggtca tccgcaaggg ctggctgact atcaataata ttggcatcat gaaagggggc
     2221 tccaaggagt actggtttgt gctgactgct gagaatctgt cctggtacaa ggatgatgag
     2281 gagaaagaga agaaatacat gctgtctgtg gacaacctca agctgcggga cgtggagaag
     2341 ggctttatgt cgagcaagca tatctttgcc ctctttaaca cggagcagag gaatgtctac
     2401 aaggattatc ggcagctgga gctagcctgt gagacacagg aggaggtgga cagctggaag
     2461 gcctccttcc tgagggctgg cgtgtaccct gagcgtgttg gggacaaaga gaaagccagc
     2521 gagaccgagg agaatggctc cgacagcttc atgcattcca tggacccaca gctggaacgg
     2581 caagtggaga ccatccggaa tcttgtggac tcatacatgg ccattgtcaa caagaccgtg
     2641 agggacctca tgcccaagac catcatgcac ctcatgatta acaataccaa ggagttcatc
     2701 ttctcggagc tgctggccaa cctgtactcg tgtggggacc agaacacgct gatggaggag
     2761 tcggcggagc aggcacagcg gcgcgacgag atgctgcgca tgtaccacgc actgaaggag
     2821 gcgctcagca tcatcggcaa catcaacacg accaccgtca gcacgcccat gcccccgccc
     2881 gtggacgact cctggctgca ggtgcagagc gtaccggccg gacgcaggtc gcccacgtcc
     2941 agccccacgc cgcagcgccg agcccccgcc gtgcccccag cccggcccgg gtcgcggggc
     3001 cctgctcctg ggcctccgcc tgctgggtcc gccctggggg gggcgccccc cgtgccctcc
     3061 aggccggggg cttcccctga ccctttcggc cctccccctc aggtgccctc gcgccccaac
     3121 cgcgccccgc ccggggtccc cagccgatcg ggtcaggcaa gtccatcccg tcctgagagc
     3181 cccaggcccc ccttcgacaa gcttcgaatt ctgcagtcga cggtaccgcg ggcccgggat
     3241 ccaccggtcg ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
     3301 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
     3361 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
     3421 gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac
     3481 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     3541 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     3601 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     3661 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc
     3721 gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc
     3781 agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     3841 ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag
     3901 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
     3961 gagctgtaca agtaaagcgg ccgcgactct agatcataat cagccatacc acatttgtag
     4021 aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga
     4081 atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     4141 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     4201 aactcatcaa tgtatcttaa ggcgtaaatt gtaagcgtta atattttgtt aaaattcgcg
     4261 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     4321 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     4381 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     4441 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     4501 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     4561 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     4621 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     4681 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     4741 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     4801 aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg
     4861 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     4921 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     4981 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     5041 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     5101 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     5161 gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     5221 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     5281 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     5341 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg
     5401 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     5461 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     5521 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     5581 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     5641 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     5701 gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc
     5761 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     5821 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     5881 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     5941 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     6001 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     6061 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     6121 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga
     6181 ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa
     6241 aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc
     6301 agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt
     6361 cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg
     6421 tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag attgatttaa
     6481 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     6541 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     6601 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     6661 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     6721 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     6781 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     6841 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     6901 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     6961 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     7021 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     7081 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     7141 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     7201 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     7261 ttcctgcgtt atcccctgat tctgtggata accgtattac cgccatgcat tagttatta
//
