LOCUS       sequence-176898-        6048 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  expression of M1 C448Y spastin in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 6048)
  AUTHORS   Solowska JM, D'Rozario M, Jean DC, Davidson MW, Marenda DR, Baas PW
  TITLE     Pathogenic mutation of spastin has gain-of-function effects on 
            microtubule dynamics.
  JOURNAL   J Neurosci. 2014 Jan 29;34(5):1856-67. doi: 
            10.1523/JNEUROSCI.3309-13.2014.
  PUBMED    24478365
REFERENCE   2  (bases 1 to 6048)
  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: 
            "10.1523/JNEUROSCI.3309-13"; journalName: "J Neurosci"; date: 
            "2014-01-29- 29"; volume: "34"; issue: "5"; pages: "1856-67"
COMMENT     Sequence Label: pCMV-Tag/WT M1
FEATURES             Location/Qualifiers
     source          1..6048
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     complement(1672..1688)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1673..1689)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(1744..1763)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1745..1763)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    2037..2158
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2074..2093)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2128..2147
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2165..2620)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2302..2323)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2514..2533
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2647..2751
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2753..3110
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2753..2944
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2776..2796)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2961..3096
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3145..3939
                     /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             3145..3939
                     /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(3199..3218)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3809..3828
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4127..4146)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4171..4218
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4547..5135
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5036..5055
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        5310..5613
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5614..5817
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5768..5788
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        5863..5881
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 gaggagctcc ggggccgcgc cagcacctgc ctcggcctcg gccccggcgc cggtgccggg
       61 cggcgaggcc gagcgcgtcc gagtcttcca caaacaggcc ttcgagtaca tctccattgc
      121 cctgcgcatc gatgaggatg agaaagcagg acagaaggag caagctgtgg aatggtataa
      181 gaaaggtatt gaagaactgg aaaaaggaat agctgttata gttacaggac aaggtgaaca
      241 gtgtgaaaga gctagacgcc ttcaagctaa aatgatgact aatttggtta tggccaagga
      301 ccgcttacaa cttctagaga agatgcaacc agttttgcca ttttccaagt cacaaacgga
      361 cgtctataat gacagtacta acttggcatg ccgcaatgga catctccagt cagaaagtgg
      421 agctgttcca aaaagaaaag accccttaac acacactagt aattcactgc ctcgttcaaa
      481 aacagttatg aaaactggat ctgcaggcct ttcaggccac catagagcac ctagttacag
      541 tggtttatcc atggtttctg gagtgaaaca gggatctggt cctgctccta ccactcataa
      601 gggtactccg aaaacaaata ggacaaataa accttctacc cctacaactg ctactcgtaa
      661 gaaaaaagac ttgaagaatt ttaggaatgt ggacagcaac cttgctaacc ttataatgaa
      721 tgaaattgtg gacaatggaa cagctgttaa atttgatgat atagctggtc aagacttggc
      781 aaaacaagca ttgcaagaaa ttgttattct tccttctctg aggcctgagt tgttcacagg
      841 gcttagagct cctgccagag ggctgttact ctttggtcca cctgggaatg ggaagacaat
      901 gctggctaaa gcagtagctg cagaatcgaa tgcaaccttc tttaatataa gtgctgcaag
      961 tttaacttca aaatacgtgg gagaaggaga gaaattggtg agggctcttt ttgctgtggc
     1021 tcgagaactt caaccttcta taatttttat agatgaagtt gatagccttt tgtgtgaaag
     1081 aagagaaggg gagcacgatg ctagtagacg cctaaaaact gaatttctaa tagaatttga
     1141 tggtgtacag tctgctggag atgacagagt acttgtaatg ggtgcaacta ataggccaca
     1201 agagcttgat gaggctgttc tcaggcgttt catcaaacgg gtatatgtgt ctttaccaaa
     1261 tgaggagaca agactacttt tgcttaaaaa tctgttatgt aaacaaggaa gtccattgac
     1321 ccaaaaagaa ctagcacaac ttgctagaat gactgatgga tactcaggaa gtgacctaac
     1381 agctttggca aaagatgcag cactgggtcc tatccgagaa ctaaaaccag aacaggtgaa
     1441 gaatatgtct gccagtgaga tgagaaatat tcgattatct gacttcactg aatccttgaa
     1501 aaaaataaaa cgcagcgtca gccctcaaac tttagaagcg tacatacgtt ggaacaagga
     1561 ctttggagat accactgttt aaggaaatac ctttgtaaac ctgcagaaca ttttacttaa
     1621 aagaggaaac acaagatccc ccgggctgca ggaattcgat atcaagctta tcgataccgt
     1681 cgacctcgag ggggggcccg gtaccttaat taattaaggt accaggtaag tgtacccaat
     1741 tcgccctata gtgagtcgta ttacaattca ctcgatcgcc cttcccaaca gttgcgcagc
     1801 ctgaatggcg aatggagatc caatttttaa gtgtataatg tgttaaacta ctgattctaa
     1861 ttgtttgtgt attttagatt cacagtccca aggctcattt caggcccctc agtcctcaca
     1921 gtctgttcat gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa
     1981 acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact
     2041 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     2101 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaac
     2161 gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
     2221 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc
     2281 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac
     2341 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca
     2401 ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg
     2461 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag
     2521 aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc
     2581 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac ttttcgggga
     2641 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     2701 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagaa tcctgaggcg
     2761 gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     2821 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     2881 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     2941 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     3001 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc
     3061 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa
     3121 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     3181 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     3241 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     3301 ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg
     3361 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     3421 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     3481 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     3541 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     3601 gtcgatcagg atgatctgga cgaagaacat caggggctcg cgccagccga actgttcgcc
     3661 aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     3721 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     3781 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagaactt
     3841 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     3901 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
     3961 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     4021 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     4081 gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa acacggaagg
     4141 agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg
     4201 gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata
     4261 ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc
     4321 ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca
     4381 tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     4441 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     4501 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     4561 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     4621 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     4681 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     4741 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     4801 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     4861 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     4921 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     4981 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     5041 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     5101 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     5161 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     5221 ctgtggataa ccgtattacc gccatgcatt agttattaat agtaatcaat tacggggtca
     5281 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
     5341 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
     5401 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
     5461 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
     5521 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
     5581 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
     5641 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
     5701 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
     5761 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctggt
     5821 ttagtgaacc gtcagatccg ctagcgatta cgccaagctc gaaattaacc ctcactaaag
     5881 ggaacaaaag ctggagtcgc ggcggcggca gtgagaggcc accatgaatt ctccgggtgg
     5941 acgagggaag aagaaaggct ccggcggcgc cagcaacccg gtgcctccca ggcctccgcc
     6001 cccttgcctg gcccccgccc ctcccgccgc cgggccggcc cctccgcc
//
