LOCUS       sequence-176270-        6193 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 6193)
  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 6193)
  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/M1C448Y
FEATURES             Location/Qualifiers
     source          1..6193
                     /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(2556..2572)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2557..2573)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(2628..2647)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2629..2647)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    2921..3042
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2958..2977)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3012..3031
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3049..3504)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3186..3207)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3398..3417
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3531..3635
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3637..3994
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3637..3828
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3660..3680)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3845..3980
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4029..4823
                     /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             4029..4823
                     /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(4083..4102)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4693..4712
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5011..5030)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5055..5102
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5431..6019
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5920..5939
                     /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 gctggagatc gcggcggcgg
      601 cagtgagagg ccaccatgaa ttctccgggt ggacgaggga agaagaaagg ctccggcggc
      661 gccagcaacc cggtgcctcc caggcctccg cccccttgcc tggcccccgc ccctcccgcc
      721 gccgggccgg cccctccgcc cgagtcgccg cataagcgga acctgtacta tttctcctac
      781 ccgctgtttg taggcttcgc gctgctgcgt ttggtcgcct tccacctggg gctcctcttc
      841 gtgtggctct gccagcgctt ctcccgcgcc ctcatggcag ccaagaggag ctccggggcc
      901 gcgccagcac ctgcctcggc ctcggccccg gcgccggtgc cgggcggcga ggccgagcgc
      961 gtccgagtct tccacaaaca ggccttcgag tacatctcca ttgccctgcg catcgatgag
     1021 gatgagaaag caggacagaa ggagcaagct gtggaatggt ataagaaagg tattgaagaa
     1081 ctggaaaaag gaatagctgt tatagttaca ggacaaggtg aacagtgtga aagagctaga
     1141 cgccttcaag ctaaaatgat gactaatttg gttatggcca aggaccgctt acaacttcta
     1201 gagaagatgc aaccagtttt gccattttcc aagtcacaaa cggacgtcta taatgacagt
     1261 actaacttgg catgccgcaa tggacatctc cagtcagaaa gtggagctgt tccaaaaaga
     1321 aaagacccct taacacacac tagtaattca ctgcctcgtt caaaaacagt tatgaaaact
     1381 ggatctgcag gcctttcagg ccaccataga gcacctagtt acagtggttt atccatggtt
     1441 tctggagtga aacagggatc tggtcctgct cctaccactc ataagggtac tccgaaaaca
     1501 aataggacaa ataaaccttc tacccctaca actgctactc gtaagaaaaa agacttgaag
     1561 aattttagga atgtggacag caaccttgct aaccttataa tgaatgaaat tgtggacaat
     1621 ggaacagctg ttaaatttga tgatatagct ggtcaagact tggcaaaaca agcattgcaa
     1681 gaaattgtta ttcttccttc tctgaggcct gagttgttca cagggcttag agctcctgcc
     1741 agagggctgt tactctttgg tccacctggg aatgggaaga caatgctggc taaagcagta
     1801 gctgcagaat cgaatgcaac cttctttaat ataagtgctg caagtttaac ttcaaaatac
     1861 gtgggagaag gagagaaatt ggtgagggct ctttttgctg tggctcgaga acttcaacct
     1921 tctataattt ttatagatga agttgatagc cttttgtatg aaagaagaga aggggagcac
     1981 gatgctagta gacgcctaaa aactgaattt ctaatagaat ttgatggtgt acagtctgct
     2041 ggagatgaca gagtacttgt aatgggtgca actaataggc cacaagagct tgatgaggct
     2101 gttctcaggc gtttcatcaa acgggtatat gtgtctttac caaatgagga gacaagacta
     2161 cttttgctta aaaatctgtt atgtaaacaa ggaagtccat tgacccaaaa agaactagca
     2221 caacttgcta gaatgactga tggatactca ggaagtgacc taacagcttt ggcaaaagat
     2281 gcagcactgg gtcctatccg agaactaaaa ccagaacagg tgaagaatat gtctgccagt
     2341 gagatgagaa atattcgatt atctgacttc actgaatcct tgaaaaaaat aaaacgcagc
     2401 gtcagccctc aaactttaga agcgtacata cgttggaaca aggactttgg agataccact
     2461 gtttaaggaa atacctttgt aaacctgcag aacattttac ttaaaagagg aaacacaaga
     2521 tcccccgggc tgcaggaatt cgatatcaag cttatcgata ccgtcgacct cgaggggggg
     2581 cccggtacct taattaatta aggtaccagg taagtgtacc caattcgccc tatagtgagt
     2641 cgtattacaa ttcactcgat cgcccttccc aacagttgcg cagcctgaat ggcgaatgga
     2701 gatccaattt ttaagtgtat aatgtgttaa actactgatt ctaattgttt gtgtatttta
     2761 gattcacagt cccaaggctc atttcaggcc cctcagtcct cacagtctgt tcatgatcat
     2821 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc
     2881 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
     2941 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     3001 gcattctagt tgtggtttgt ccaaactcat caatgtatct taacgcgtaa attgtaagcg
     3061 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     3121 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     3181 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     3241 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
     3301 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     3361 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     3421 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     3481 ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     3541 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     3601 gataaatgct tcaataatat tgaaaaagga agaatcctga ggcggaaaga accagctgtg
     3661 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3721 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     3781 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     3841 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     3901 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
     3961 aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat
     4021 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     4081 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     4141 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     4201 atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     4261 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     4321 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     4381 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     4441 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     4501 tggacgaaga acatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca
     4561 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     4621 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     4681 atcaggacat agcgttggct acccgtgata ttgctgaaga acttggcggc gaatgggctg
     4741 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     4801 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac
     4861 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     4921 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     4981 gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg
     5041 aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt
     5101 tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca
     5161 ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga
     5221 aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc
     5281 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     5341 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     5401 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     5461 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     5521 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     5581 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     5641 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     5701 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     5761 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     5821 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     5881 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     5941 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     6001 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     6061 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     6121 taccgccatg cattagttat taatagtaat caattacggg gtcattagtt catagcccat
     6181 atatggagtt ccg
//
