LOCUS       sequence-389676-        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 E720R
FEATURES             Location/Qualifiers
     source          1..7319
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(76..95)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    120..167
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      496..1084
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     985..1004
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        1259..1562
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1563..1766
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1717..1737
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      4457..4466
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4463..4465,4466..4468,4469..5182)
                     /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(4463..4465,4466..4468,4469..5179)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4508..4529)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4769..4788)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5116..5137
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    5305..5426
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5342..5361)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5396..5415
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5433..5888)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5570..5591)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5782..5801
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5915..6019
                     /gene="bla"
                     /label=AmpR promoter
     promoter        6021..6378
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6021..6212
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6044..6064)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6229..6364
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6413..7207
                     /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             6413..7207
                     /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(6467..6486)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7077..7096
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
ORIGIN
        1 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
       61 tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac
      121 ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata aacgcggggt
      181 tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc
      241 ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc
      301 agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata tactttagat
      361 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
      421 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
      481 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
      541 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
      601 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
      661 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
      721 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
      781 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
      841 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
      901 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
      961 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     1021 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     1081 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     1141 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcatta
     1201 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
     1261 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
     1321 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
     1381 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
     1441 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
     1501 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
     1561 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
     1621 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
     1681 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
     1741 ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc tagcgctacc
     1801 ggactcagat cctgccatgg gcaaccgcgg catggaagat ctcatcccgc tggtcaaccg
     1861 gctgcaagac gccttctctg ccatcggcca gaacgcggac ctcgacctgc cgcagatcgc
     1921 tgtggtgggc ggccagagcg ccggcaagag ctcggtgctc gagaatttcg taggcaggga
     1981 cttcttgcct cgaggatctg gcattgttac ccgacgtccc ctggtcttgc agctggtcaa
     2041 tgcaaccaca gaatatgccg agttcctgca ctgcaaggga aagaaattca ccgacttcga
     2101 ggaggtgcgc cttgagatcg aggccgagac cgacagggtc accggcacca acaagggcat
     2161 ctcgccggtg cctatcaacc tccgcgtcta ctcgccgcac gtgctgaacc tgaccctggt
     2221 ggacctgccc ggaatgacca aggtcccggt gggggaccaa cctcccgaca tcgagttcca
     2281 gatccgagac atgcttatgc agtttgtcac caaggagaac tgcctcatcc tggccgtgtc
     2341 ccccgccaac tctgacctgg ccaattctga cgccctcaag gtcgccaagg aggtggaccc
     2401 ccagggccag cgcaccatcg gggtcatcac caagctggac ctgatggacg agggcacaga
     2461 tgcccgtgat gtgctggaga acaagctgct ccccctgcgc agaggctaca ttggagtggt
     2521 gaaccggagc cagaaggaca ttgatggcaa gaaggacatt accgccgcct tggctgctga
     2581 acgaaagttc ttcctctccc atccatctta tcgccacttg gctgacgaga tgggcacgcc
     2641 ctacctgcag aaggtcctca atcagcaact gacgaaccac atccgggaca cactgccggg
     2701 gctgcggaac aagctgcaga gccagctact gtccattgag aaggaggtgg aggaatacaa
     2761 gaacttccgc cctgatgacc cagctcgcaa gaccaaggcc ctgctgcaga tggtccagca
     2821 gtttgccgta gactttgaga agcgcattga gggctcagga gatcagatcg acacctacga
     2881 actgtcaggg ggagcccgca ttaaccgaat cttccacgag cgcttccctt tcgagctggt
     2941 caagatggag tttgatgaga aggaactccg aagggagatc agctatgcta tcaagaatat
     3001 ccatggcatt agaacggggc tgtttacccc agacatggcc tttgagacca ttgtgaaaaa
     3061 gcaggtgaag aagatccgag aaccgtgtct caagtgtgtg gacatggtta tctcggagct
     3121 aatcagcacc gttagacagt gcaccaagaa gctccagcag tacccgcggc tacgggagga
     3181 gatggagcgc atcgtgacca cccacatccg ggagcgcgag ggccgcacta aggagcaggt
     3241 catgcttctc atcgatatcg agctggctta catgaacacc aaccatgagg acttcatagg
     3301 ctttgccaat gctcagcaga ggagcaacca gatgaacaag aagaagactt cagggaacca
     3361 ggatgagatt ctggtcatcc gcaagggctg gctgactatc aataatattg gcatcatgaa
     3421 agggggctcc aaggagtact ggtttgtgct gactgctgag aatctgtcct ggtacaagga
     3481 tgatgaggag aaagagaaga aatacatgct gtctgtggac aacctcaagc tgcgggacgt
     3541 ggagaagggc tttatgtcga gcaagcatat ctttgccctc tttaacacgg agcagaggaa
     3601 tgtctacaag gattatcggc agctggagct agcctgtgag acacaggagg aggtggacag
     3661 ctggaaggcc tccttcctga gggctggcgt gtaccctgag cgtgttgggg acaaagagaa
     3721 agccagcgag accgaggaga atggctccga cagcttcatg cattccatgg acccacagct
     3781 ggaacggcaa gtggagacca tccggaatct tgtggactca tacatggcca ttgtcaacaa
     3841 gaccgtgagg gacctcatgc ccaagaccat catgcacctc atgattaaca ataccaagga
     3901 gttcatcttc tcggagctgc tggccaacct gtactcgtgt ggggaccaga acacgctgat
     3961 ggaggagtcg gcgaggcagg cacagcggcg cgacgagatg ctgcgcatgt accacgcact
     4021 gaaggaggcg ctcagcatca tcggcaacat caacacgacc accgtcagca cgcccatgcc
     4081 cccgcccgtg gacgactcct ggctgcaggt gcagagcgta ccggccggac gcaggtcgcc
     4141 cacgtccagc cccacgccgc agcgccgagc ccccgccgtg cccccagccc ggcccgggtc
     4201 gcggggccct gctcctgggc ctccgcctgc tgggtccgcc ctgggggggg cgccccccgt
     4261 gccctccagg ccgggggctt cccctgaccc tttcggccct ccccctcagg tgccctcgcg
     4321 ccccaaccgc gccccgcccg gggtccccag ccgatcgggt caggcaagtc catcccgtcc
     4381 tgagagcccc aggcccccct tcgacaagct tcgaattctg cagtcgacgg taccgcgggc
     4441 ccgggatcca ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt
     4501 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
     4561 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa
     4621 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag
     4681 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
     4741 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
     4801 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
     4861 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat
     4921 catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga
     4981 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
     5041 cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa
     5101 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg
     5161 catggacgag ctgtacaagt aaagcggccg cgactctaga tcataatcag ccataccaca
     5221 tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat
     5281 aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa
     5341 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     5401 ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta agcgttaata ttttgttaaa
     5461 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
     5521 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa
     5581 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     5641 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
     5701 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
     5761 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
     5821 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
     5881 gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     5941 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     6001 atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta
     6061 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     6121 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     6181 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     6241 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     6301 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     6361 ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga
     6421 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     6481 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     6541 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga
     6601 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     6661 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     6721 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     6781 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     6841 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     6901 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga
     6961 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     7021 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     7081 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     7141 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     7201 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca
     7261 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccg
//
