LOCUS       sequence-176897-        6192 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of M1 C448Y spastin in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6192)
  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 6192)
  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..6192
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        554..572
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2555..2571)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2556..2572)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(2627..2646)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2628..2646)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    2920..3041
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2957..2976)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3011..3030
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3048..3503)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3185..3206)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3397..3416
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3530..3634
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3636..3993
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3636..3827
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3659..3679)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3844..3979
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4028..4822
                     /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             4028..4822
                     /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(4082..4101)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4692..4711
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5010..5029)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5054..5101
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5430..6018
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5919..5938
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgatt
      541 acgccaagct cgaaattaac cctcactaaa gggaacaaaa gctggagtcg cggcggcggc
      601 agtgagaggc caccatgaat tctccgggtg gacgagggaa gaagaaaggc tccggcggcg
      661 ccagcaaccc ggtgcctccc aggcctccgc ccccttgcct ggcccccgcc cctcccgccg
      721 ccgggccggc ccctccgccc gagtcgccgc ataagcggaa cctgtactat ttctcctacc
      781 cgctgtttgt aggcttcgcg ctgctgcgtt tggtcgcctt ccacctgggg ctcctcttcg
      841 tgtggctctg ccagcgcttc tcccgcgccc tcatggcagc caagaggagc tccggggccg
      901 cgccagcacc tgcctcggcc tcggccccgg cgccggtgcc gggcggcgag gccgagcgcg
      961 tccgagtctt ccacaaacag gccttcgagt acatctccat tgccctgcgc atcgatgagg
     1021 atgagaaagc aggacagaag gagcaagctg tggaatggta taagaaaggt attgaagaac
     1081 tggaaaaagg aatagctgtt atagttacag gacaaggtga acagtgtgaa agagctagac
     1141 gccttcaagc taaaatgatg actaatttgg ttatggccaa ggaccgctta caacttctag
     1201 agaagatgca accagttttg ccattttcca agtcacaaac ggacgtctat aatgacagta
     1261 ctaacttggc atgccgcaat ggacatctcc agtcagaaag tggagctgtt ccaaaaagaa
     1321 aagacccctt aacacacact agtaattcac tgcctcgttc aaaaacagtt atgaaaactg
     1381 gatctgcagg cctttcaggc caccatagag cacctagtta cagtggttta tccatggttt
     1441 ctggagtgaa acagggatct ggtcctgctc ctaccactca taagggtact ccgaaaacaa
     1501 ataggacaaa taaaccttct acccctacaa ctgctactcg taagaaaaaa gacttgaaga
     1561 attttaggaa tgtggacagc aaccttgcta accttataat gaatgaaatt gtggacaatg
     1621 gaacagctgt taaatttgat gatatagctg gtcaagactt ggcaaaacaa gcattgcaag
     1681 aaattgttat tcttccttct ctgaggcctg agttgttcac agggcttaga gctcctgcca
     1741 gagggctgtt actctttggt ccacctggga atgggaagac aatgctggct aaagcagtag
     1801 ctgcagaatc gaatgcaacc ttctttaata taagtgctgc aagtttaact tcaaaatacg
     1861 tgggagaagg agagaaattg gtgagggctc tttttgctgt ggctcgagaa cttcaacctt
     1921 ctataatttt tatagatgaa gttgatagcc ttttgtgtga aagaagagaa ggggagcacg
     1981 atgctagtag acgcctaaaa actgaatttc taatagaatt tgatggtgta cagtctgctg
     2041 gagatgacag agtacttgta atgggtgcaa ctaataggcc acaagagctt gatgaggctg
     2101 ttctcaggcg tttcatcaaa cgggtatatg tgtctttacc aaatgaggag acaagactac
     2161 ttttgcttaa aaatctgtta tgtaaacaag gaagtccatt gacccaaaaa gaactagcac
     2221 aacttgctag aatgactgat ggatactcag gaagtgacct aacagctttg gcaaaagatg
     2281 cagcactggg tcctatccga gaactaaaac cagaacaggt gaagaatatg tctgccagtg
     2341 agatgagaaa tattcgatta tctgacttca ctgaatcctt gaaaaaaata aaacgcagcg
     2401 tcagccctca aactttagaa gcgtacatac gttggaacaa ggactttgga gataccactg
     2461 tttaaggaaa tacctttgta aacctgcaga acattttact taaaagagga aacacaagat
     2521 cccccgggct gcaggaattc gatatcaagc ttatcgatac cgtcgacctc gagggggggc
     2581 ccggtacctt aattaattaa ggtaccaggt aagtgtaccc aattcgccct atagtgagtc
     2641 gtattacaat tcactcgatc gcccttccca acagttgcgc agcctgaatg gcgaatggag
     2701 atccaatttt taagtgtata atgtgttaaa ctactgattc taattgtttg tgtattttag
     2761 attcacagtc ccaaggctca tttcaggccc ctcagtcctc acagtctgtt catgatcata
     2821 atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc
     2881 ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat
     2941 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     3001 cattctagtt gtggtttgtc caaactcatc aatgtatctt aacgcgtaaa ttgtaagcgt
     3061 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
     3121 ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt
     3181 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
     3241 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt
     3301 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
     3361 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
     3421 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
     3481 taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     3541 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     3601 ataaatgctt caataatatt gaaaaaggaa gaatcctgag gcggaaagaa ccagctgtgg
     3661 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3721 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     3781 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     3841 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     3901 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     3961 ggaggctttt ttggaggcct aggcttttgc aaagatcgat caagagacag gatgaggatc
     4021 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     4081 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     4141 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     4201 tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     4261 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     4321 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     4381 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     4441 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     4501 ggacgaagaa catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgagcat
     4561 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     4621 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     4681 tcaggacata gcgttggcta cccgtgatat tgctgaagaa cttggcggcg aatgggctga
     4741 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     4801 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
     4861 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     4921 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     4981 ttcttcgccc accctagggg gaggctaact gaaacacgga aggagacaat accggaagga
     5041 acccgcgcta tgacggcaat aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt
     5101 cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat
     5161 tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa
     5221 ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc aggttactca
     5281 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     5341 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     5401 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     5461 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     5521 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     5581 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     5641 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     5701 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     5761 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     5821 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     5881 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     5941 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     6001 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     6061 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     6121 accgccatgc attagttatt aatagtaatc aattacgggg tcattagttc atagcccata
     6181 tatggagttc cg
//
