LOCUS       sequence-389681-        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 D291R
FEATURES             Location/Qualifiers
     source          1..7319
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    44..165
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(81..100)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     135..154
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(172..627)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(309..330)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     521..540
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        654..758
                     /gene="bla"
                     /label=AmpR promoter
     promoter        760..1117
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        760..951
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(783..803)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      968..1103
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             1152..1946
                     /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             1152..1946
                     /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(1206..1225)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1816..1835
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(2134..2153)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2178..2225
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      2554..3142
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3043..3062
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        3317..3620
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3621..3824
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3775..3795
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      6515..6524
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(6521..6523,6524..6526,6527..7240)
                     /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(6521..6523,6524..6526,6527..7237)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(6566..6587)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6827..6846)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7174..7195
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc
       61 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
      121 actgcattct agttgtggtt tgtccaaact catcaatgta tcttaaggcg taaattgtaa
      181 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
      241 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
      301 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
      361 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
      421 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
      481 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
      541 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
      601 cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa
      661 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
      721 cctgataaat gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct
      781 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
      841 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
      901 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
      961 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     1021 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     1081 gtgaggaggc ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag
     1141 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     1201 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     1261 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     1321 tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     1381 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     1441 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     1501 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     1561 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     1621 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga
     1681 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     1741 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     1801 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     1861 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     1921 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     1981 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     2041 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     2101 ggagttcttc gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga
     2161 aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt
     2221 tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc
     2281 ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg
     2341 tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta
     2401 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     2461 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     2521 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     2581 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     2641 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     2701 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     2761 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     2821 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     2881 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     2941 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     3001 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     3061 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     3121 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     3181 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     3241 tattaccgcc atgcattagt tattaatagt aatcaattac ggggtcatta gttcatagcc
     3301 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
     3361 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
     3421 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
     3481 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
     3541 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
     3601 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
     3661 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
     3721 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
     3781 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc
     3841 agatccgcta gcgctaccgg actcagatcc tgccatgggc aaccgcggca tggaagatct
     3901 catcccgctg gtcaaccggc tgcaagacgc cttctctgcc atcggccaga acgcggacct
     3961 cgacctgccg cagatcgctg tggtgggcgg ccagagcgcc ggcaagagct cggtgctcga
     4021 gaatttcgta ggcagggact tcttgcctcg aggatctggc attgttaccc gacgtcccct
     4081 ggtcttgcag ctggtcaatg caaccacaga atatgccgag ttcctgcact gcaagggaaa
     4141 gaaattcacc gacttcgagg aggtgcgcct tgagatcgag gccgagaccg acagggtcac
     4201 cggcaccaac aagggcatct cgccggtgcc tatcaacctc cgcgtctact cgccgcacgt
     4261 gctgaacctg accctggtgg acctgcccgg aatgaccaag gtcccggtgg gggaccaacc
     4321 tcccgacatc gagttccaga tccgagacat gcttatgcag tttgtcacca aggagaactg
     4381 cctcatcctg gccgtgtccc ccgccaactc tgacctggcc aattctgacg ccctcaaggt
     4441 cgccaaggag gtggaccccc agggccagcg caccatcggg gtcatcacca agctggacct
     4501 gatggacgag ggcacagatg cccgtgatgt gctggagaac aagctgctcc ccctgcgcag
     4561 aggctacatt ggagtggtga accggagcca gaaggacatt gatggcaaga aggacattac
     4621 cgccgccttg gctgctgaac gaaagttctt cctctcccat ccatcttatc gccacttggc
     4681 tgacgagatg ggcacgccct acctgcagaa ggtcctcaat cagcaactga cgaaccacat
     4741 ccggaggaca ctgccggggc tgcggaacaa gctgcagagc cagctactgt ccattgagaa
     4801 ggaggtggag gaatacaaga acttccgccc tgatgaccca gctcgcaaga ccaaggccct
     4861 gctgcagatg gtccagcagt ttgccgtaga ctttgagaag cgcattgagg gctcaggaga
     4921 tcagatcgac acctacgaac tgtcaggggg agcccgcatt aaccgaatct tccacgagcg
     4981 cttccctttc gagctggtca agatggagtt tgatgagaag gaactccgaa gggagatcag
     5041 ctatgctatc aagaatatcc atggcattag aacggggctg tttaccccag acatggcctt
     5101 tgagaccatt gtgaaaaagc aggtgaagaa gatccgagaa ccgtgtctca agtgtgtgga
     5161 catggttatc tcggagctaa tcagcaccgt tagacagtgc accaagaagc tccagcagta
     5221 cccgcggcta cgggaggaga tggagcgcat cgtgaccacc cacatccggg agcgcgaggg
     5281 ccgcactaag gagcaggtca tgcttctcat cgatatcgag ctggcttaca tgaacaccaa
     5341 ccatgaggac ttcataggct ttgccaatgc tcagcagagg agcaaccaga tgaacaagaa
     5401 gaagacttca gggaaccagg atgagattct ggtcatccgc aagggctggc tgactatcaa
     5461 taatattggc atcatgaaag ggggctccaa ggagtactgg tttgtgctga ctgctgagaa
     5521 tctgtcctgg tacaaggatg atgaggagaa agagaagaaa tacatgctgt ctgtggacaa
     5581 cctcaagctg cgggacgtgg agaagggctt tatgtcgagc aagcatatct ttgccctctt
     5641 taacacggag cagaggaatg tctacaagga ttatcggcag ctggagctag cctgtgagac
     5701 acaggaggag gtggacagct ggaaggcctc cttcctgagg gctggcgtgt accctgagcg
     5761 tgttggggac aaagagaaag ccagcgagac cgaggagaat ggctccgaca gcttcatgca
     5821 ttccatggac ccacagctgg aacggcaagt ggagaccatc cggaatcttg tggactcata
     5881 catggccatt gtcaacaaga ccgtgaggga cctcatgccc aagaccatca tgcacctcat
     5941 gattaacaat accaaggagt tcatcttctc ggagctgctg gccaacctgt actcgtgtgg
     6001 ggaccagaac acgctgatgg aggagtcggc ggagcaggca cagcggcgcg acgagatgct
     6061 gcgcatgtac cacgcactga aggaggcgct cagcatcatc ggcaacatca acacgaccac
     6121 cgtcagcacg cccatgcccc cgcccgtgga cgactcctgg ctgcaggtgc agagcgtacc
     6181 ggccggacgc aggtcgccca cgtccagccc cacgccgcag cgccgagccc ccgccgtgcc
     6241 cccagcccgg cccgggtcgc ggggccctgc tcctgggcct ccgcctgctg ggtccgccct
     6301 ggggggggcg ccccccgtgc cctccaggcc gggggcttcc cctgaccctt tcggccctcc
     6361 ccctcaggtg ccctcgcgcc ccaaccgcgc cccgcccggg gtccccagcc gatcgggtca
     6421 ggcaagtcca tcccgtcctg agagccccag gccccccttc gacaagcttc gaattctgca
     6481 gtcgacggta ccgcgggccc gggatccacc ggtcgccacc atggtgagca agggcgagga
     6541 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
     6601 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt
     6661 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta
     6721 cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc
     6781 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
     6841 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     6901 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa
     6961 cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa
     7021 gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac
     7081 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc
     7141 cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc
     7201 cgccgggatc actctcggca tggacgagct gtacaagtaa agcggccgcg actctagatc
     7261 ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacct
//
