LOCUS       sequence-389671-        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 D406A
FEATURES             Location/Qualifiers
     source          1..7319
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     416..435
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(734..753)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    778..825
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1154..1742
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1643..1662
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        1917..2220
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2221..2424
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2375..2395
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      5115..5124
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(5121..5123,5124..5126,5127..5840)
                     /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(5121..5123,5124..5126,5127..5837)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5166..5187)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5427..5446)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5774..5795
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    5963..6084
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6000..6019)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6054..6073
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(6091..6546)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6228..6249)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6440..6459
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6573..6677
                     /gene="bla"
                     /label=AmpR promoter
     promoter        6679..7036
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6679..6870
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6702..6722)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6887..7022
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             join(7071..7319,1..546)
                     /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             join(7071..7319,1..546)
                     /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(7125..7144)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
       61 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
      121 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
      181 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
      241 gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat
      301 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
      361 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
      421 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
      481 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
      541 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
      601 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
      661 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta
      721 gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg
      781 caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc
      841 ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc
      901 gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag
      961 ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg
     1021 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     1081 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     1141 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     1201 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     1261 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     1321 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     1381 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     1441 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     1501 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     1561 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     1621 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     1681 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     1741 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     1801 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcattagt
     1861 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
     1921 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
     1981 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
     2041 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
     2101 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
     2161 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
     2221 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
     2281 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
     2341 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
     2401 tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta gcgctaccgg
     2461 actcagatcc tgccatgggc aaccgcggca tggaagatct catcccgctg gtcaaccggc
     2521 tgcaagacgc cttctctgcc atcggccaga acgcggacct cgacctgccg cagatcgctg
     2581 tggtgggcgg ccagagcgcc ggcaagagct cggtgctcga gaatttcgta ggcagggact
     2641 tcttgcctcg aggatctggc attgttaccc gacgtcccct ggtcttgcag ctggtcaatg
     2701 caaccacaga atatgccgag ttcctgcact gcaagggaaa gaaattcacc gacttcgagg
     2761 aggtgcgcct tgagatcgag gccgagaccg acagggtcac cggcaccaac aagggcatct
     2821 cgccggtgcc tatcaacctc cgcgtctact cgccgcacgt gctgaacctg accctggtgg
     2881 acctgcccgg aatgaccaag gtcccggtgg gggaccaacc tcccgacatc gagttccaga
     2941 tccgagacat gcttatgcag tttgtcacca aggagaactg cctcatcctg gccgtgtccc
     3001 ccgccaactc tgacctggcc aattctgacg ccctcaaggt cgccaaggag gtggaccccc
     3061 agggccagcg caccatcggg gtcatcacca agctggacct gatggacgag ggcacagatg
     3121 cccgtgatgt gctggagaac aagctgctcc ccctgcgcag aggctacatt ggagtggtga
     3181 accggagcca gaaggacatt gatggcaaga aggacattac cgccgccttg gctgctgaac
     3241 gaaagttctt cctctcccat ccatcttatc gccacttggc tgaccgtatg ggcacgccct
     3301 acctgcagaa ggtcctcaat cagcaactga cgaaccacat ccgggacaca ctgccggggc
     3361 tgcggaacaa gctgcagagc cagctactgt ccattgagaa ggaggtggag gaatacaaga
     3421 acttccgccc tgatgaccca gctcgcaaga ccaaggccct gctgcagatg gtccagcagt
     3481 ttgccgtaga ctttgagaag cgcattgagg gctcaggaga tcagatcgac acctacgaac
     3541 tgtcaggggg agcccgcatt aaccgaatct tccacgagcg cttccctttc gagctggtca
     3601 agatggagtt tgatgagaag gaactccgaa gggagatcag ctatgctatc aagaatatcc
     3661 atggcattag aacggggctg tttaccccag ccatggcctt tgagaccatt gtgaaaaagc
     3721 aggtgaagaa gatccgagaa ccgtgtctca agtgtgtgga catggttatc tcggagctaa
     3781 tcagcaccgt tagacagtgc accaagaagc tccagcagta cccgcggcta cgggaggaga
     3841 tggagcgcat cgtgaccacc cacatccggg agcgcgaggg ccgcactaag gagcaggtca
     3901 tgcttctcat cgatatcgag ctggcttaca tgaacaccaa ccatgaggac ttcataggct
     3961 ttgccaatgc tcagcagagg agcaaccaga tgaacaagaa gaagacttca gggaaccagg
     4021 atgagattct ggtcatccgc aagggctggc tgactatcaa taatattggc atcatgaaag
     4081 ggggctccaa ggagtactgg tttgtgctga ctgctgagaa tctgtcctgg tacaaggatg
     4141 atgaggagaa agagaagaaa tacatgctgt ctgtggacaa cctcaagctg cgggacgtgg
     4201 agaagggctt tatgtcgagc aagcatatct ttgccctctt taacacggag cagaggaatg
     4261 tctacaagga ttatcggcag ctggagctag cctgtgagac acaggaggag gtggacagct
     4321 ggaaggcctc cttcctgagg gctggcgtgt accctgagcg tgttggggac aaagagaaag
     4381 ccagcgagac cgaggagaat ggctccgaca gcttcatgca ttccatggac ccacagctgg
     4441 aacggcaagt ggagaccatc cggaatcttg tggactcata catggccatt gtcaacaaga
     4501 ccgtgaggga cctcatgccc aagaccatca tgcacctcat gattaacaat accaaggagt
     4561 tcatcttctc ggagctgctg gccaacctgt actcgtgtgg ggaccagaac acgctgatgg
     4621 aggagtcggc ggagcaggca cagcggcgcg acgagatgct gcgcatgtac cacgcactga
     4681 aggaggcgct cagcatcatc ggcaacatca acacgaccac cgtcagcacg cccatgcccc
     4741 cgcccgtgga cgactcctgg ctgcaggtgc agagcgtacc ggccggacgc aggtcgccca
     4801 cgtccagccc cacgccgcag cgccgagccc ccgccgtgcc cccagcccgg cccgggtcgc
     4861 ggggccctgc tcctgggcct ccgcctgctg ggtccgccct ggggggggcg ccccccgtgc
     4921 cctccaggcc gggggcttcc cctgaccctt tcggccctcc ccctcaggtg ccctcgcgcc
     4981 ccaaccgcgc cccgcccggg gtccccagcc gatcgggtca ggcaagtcca tcccgtcctg
     5041 agagccccag gccccccttc gacaagcttc gaattctgca gtcgacggta ccgcgggccc
     5101 gggatccacc ggtcgccacc atggtgagca agggcgagga gctgttcacc ggggtggtgc
     5161 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
     5221 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc
     5281 tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc
     5341 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
     5401 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
     5461 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
     5521 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca
     5581 tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg
     5641 acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
     5701 tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg
     5761 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca
     5821 tggacgagct gtacaagtaa agcggccgcg actctagatc ataatcagcc ataccacatt
     5881 tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa
     5941 aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag
     6001 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     6061 gtccaaactc atcaatgtat cttaaggcgt aaattgtaag cgttaatatt ttgttaaaat
     6121 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     6181 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     6241 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     6301 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     6361 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     6421 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     6481 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     6541 gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     6601 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     6661 attgaaaaag gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg
     6721 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     6781 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     6841 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     6901 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     6961 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     7021 cctaggcttt tgcaaagatc gatcaagaga caggatgagg atcgtttcgc atgattgaac
     7081 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     7141 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     7201 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caagacgagg
     7261 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgtt
//
