LOCUS       sequence-389663-        7319 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid of GFP-chimera proteins..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7319)
  AUTHORS   Jimah JR, Kundu N, Stanton AE, Sochacki KA, Canagarajah B, Chan L, 
            Strub MP, Wang H, Taraska JW, Hinshaw JE
  TITLE     Cryo-EM structures of membrane-bound dynamin in a post-hydrolysis 
            state primed for membrane fission.
  JOURNAL   Dev Cell. 2024 Apr 23:S1534-5807(24)00233-8. doi: 
            10.1016/j.devcel.2024.04.008.
  PUBMED    38663399
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"; journalName: "Dev Cell. 
            2024 Apr 23:S1534-5807(24)00233-8. doi: 
            10.1016/j.devcel.2024.04.008."
COMMENT     Sequence Label: Dyn1 R271E
FEATURES             Location/Qualifiers
     source          1..7319
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      2672..2681
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2678..2680,2681..2683,2684..3397)
                     /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(2678..2680,2681..2683,2684..3394)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2723..2744)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2984..3003)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3331..3352
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3520..3641
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3557..3576)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3611..3630
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3648..4103)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3785..3806)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3997..4016
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4130..4234
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4236..4593
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4236..4427
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4259..4279)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4444..4579
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4628..5422
                     /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             4628..5422
                     /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(4682..4701)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5292..5311
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5610..5629)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5654..5701
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6030..6618
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6519..6538
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        6793..7096
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7097..7300
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7251..7271
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 tccgctagcg ctaccggact cagatcctgc catgggcaac cgcggcatgg aagatctcat
       61 cccgctggtc aaccggctgc aagacgcctt ctctgccatc ggccagaacg cggacctcga
      121 cctgccgcag atcgctgtgg tgggcggcca gagcgccggc aagagctcgg tgctcgagaa
      181 tttcgtaggc agggacttct tgcctcgagg atctggcatt gttacccgac gtcccctggt
      241 cttgcagctg gtcaatgcaa ccacagaata tgccgagttc ctgcactgca agggaaagaa
      301 attcaccgac ttcgaggagg tgcgccttga gatcgaggcc gagaccgaca gggtcaccgg
      361 caccaacaag ggcatctcgc cggtgcctat caacctccgc gtctactcgc cgcacgtgct
      421 gaacctgacc ctggtggacc tgcccggaat gaccaaggtc ccggtggggg accaacctcc
      481 cgacatcgag ttccagatcc gagacatgct tatgcagttt gtcaccaagg agaactgcct
      541 catcctggcc gtgtcccccg ccaactctga cctggccaat tctgacgccc tcaaggtcgc
      601 caaggaggtg gacccccagg gccagcgcac catcggggtc atcaccaagc tggacctgat
      661 ggacgagggc acagatgccc gtgatgtgct ggagaacaag ctgctccccc tgcgcagagg
      721 ctacattgga gtggtgaacc ggagccagaa ggacattgat ggcaagaagg acattaccgc
      781 cgccttggct gctgaacgaa agttcttcct ctcccatcca tcttatcgcc acttggctga
      841 cgagatgggc acgccctacc tgcagaaggt cctcaatcag caactgacga accacatccg
      901 ggacacactg ccggggctgc ggaacaagct gcagagccag ctactgtcca ttgagaagga
      961 ggtggaggaa tacaagaact tccgccctga tgacccagct cgcaagacca aggccctgct
     1021 gcagatggtc cagcagtttg ccgtagactt tgagaagcgc attgagggct caggagatca
     1081 gatcgacacc tacgaactgt cagggggagc ccgcattaac cgaatcttcc acgagcgctt
     1141 ccctttcgag ctggtcaaga tggagtttga tgagaaggaa ctccgaaggg agatcagcta
     1201 tgctatcaag aatatccatg gcattagaac ggggctgttt accccagaca tggcctttga
     1261 gaccattgtg aaaaagcagg tgaagaagat ccgagaaccg tgtctcaagt gtgtggacat
     1321 ggttatctcg gagctaatca gcaccgttag acagtgcacc aagaagctcc agcagtaccc
     1381 gcggctacgg gaggagatgg agcgcatcgt gaccacccac atccgggagc gcgagggccg
     1441 cactaaggag caggtcatgc ttctcatcga tatcgagctg gcttacatga acaccaacca
     1501 tgaggacttc ataggctttg ccaatgctca gcagaggagc aaccagatga acaagaagaa
     1561 gacttcaggg aaccaggatg agattctggt catccgcaag ggctggctga ctatcaataa
     1621 tattggcatc atgaaagggg gctccaagga gtactggttt gtgctgactg ctgagaatct
     1681 gtcctggtac aaggatgatg aggagaaaga gaagaaatac atgctgtctg tggacaacct
     1741 caagctgcgg gacgtggaga agggctttat gtcgagcaag catatctttg ccctctttaa
     1801 cacggagcag aggaatgtct acaaggatta tcggcagctg gagctagcct gtgagacaca
     1861 ggaggaggtg gacagctgga aggcctcctt cctgagggct ggcgtgtacc ctgagcgtgt
     1921 tggggacaaa gagaaagcca gcgagaccga ggagaatggc tccgacagct tcatgcattc
     1981 catggaccca cagctggaac ggcaagtgga gaccatccgg aatcttgtgg actcatacat
     2041 ggccattgtc aacaagaccg tgagggacct catgcccaag accatcatgc acctcatgat
     2101 taacaatacc aaggagttca tcttctcgga gctgctggcc aacctgtact cgtgtgggga
     2161 ccagaacacg ctgatggagg agtcggcgga gcaggcacag cggcgcgacg agatgctgcg
     2221 catgtaccac gcactgaagg aggcgctcag catcatcggc aacatcaaca cgaccaccgt
     2281 cagcacgccc atgcccccgc ccgtggacga ctcctggctg caggtgcaga gcgtaccggc
     2341 cggacgcagg tcgcccacgt ccagccccac gccgcagcgc cgagcccccg ccgtgccccc
     2401 agcccggccc gggtcgcggg gccctgctcc tgggcctccg cctgctgggt ccgccctggg
     2461 gggggcgccc cccgtgccct ccaggccggg ggcttcccct gaccctttcg gccctccccc
     2521 tcaggtgccc tcgcgcccca accgcgcccc gcccggggtc cccagccgat cgggtcaggc
     2581 aagtccatcc cgtcctgaga gccccaggcc ccccttcgac aagcttcgaa ttctgcagtc
     2641 gacggtaccg cgggcccggg atccaccggt cgccaccatg gtgagcaagg gcgaggagct
     2701 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt
     2761 cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat
     2821 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg
     2881 cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc
     2941 catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa
     3001 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
     3061 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag
     3121 ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat
     3181 ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc
     3241 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct
     3301 gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc
     3361 cgggatcact ctcggcatgg acgagctgta caagtaaagc ggccgcgact ctagatcata
     3421 atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc
     3481 ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat
     3541 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     3601 cattctagtt gtggtttgtc caaactcatc aatgtatctt aaggcgtaaa ttgtaagcgt
     3661 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
     3721 ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt
     3781 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
     3841 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt
     3901 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
     3961 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
     4021 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
     4081 taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     4141 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     4201 ataaatgctt caataatatt gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg
     4261 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     4321 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     4381 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     4441 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     4501 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     4561 ggaggctttt ttggaggcct aggcttttgc aaagatcgat caagagacag gatgaggatc
     4621 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     4681 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     4741 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     4801 tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     4861 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     4921 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     4981 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     5041 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     5101 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgagcat
     5161 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     5221 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     5281 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     5341 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     5401 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
     5461 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     5521 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     5581 ttcttcgccc accctagggg gaggctaact gaaacacgga aggagacaat accggaagga
     5641 acccgcgcta tgacggcaat aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt
     5701 cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat
     5761 tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa
     5821 ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc aggttactca
     5881 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     5941 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     6001 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     6061 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     6121 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     6181 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     6241 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     6301 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     6361 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     6421 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     6481 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     6541 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     6601 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     6661 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     6721 accgccatgc attagttatt aatagtaatc aattacgggg tcattagttc atagcccata
     6781 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
     6841 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
     6901 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
     6961 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
     7021 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
     7081 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
     7141 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
     7201 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
     7261 cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcaga
//
