LOCUS       sequence-181053-        6194 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of M1 N184X spastin in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6194)
  AUTHORS   Solowska JM, Rao AN, Baas PW
  TITLE     Studies on truncating mutations of SPAST associated with Hereditary 
            Spastic Paraplegia indicate greater accumulation and toxicity of the
            M1 isoform of spastin.
  JOURNAL   Mol Biol Cell. 2017 May 11. pii: mbc.E17-01-0047. doi: 
            10.1091/mbc.E17-01-0047.
  PUBMED    28495799
REFERENCE   2  (bases 1 to 6194)
  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: "Mol Biol 
            Cell. 2017 May 11. pii: mbc.E17-01-0047. doi: 
            10.1091/mbc.E17-01-0047."
COMMENT     Sequence Label: pCMV-Tag/M1 N184X
FEATURES             Location/Qualifiers
     source          1..6194
                     /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(2557..2573)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2558..2574)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(2629..2648)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2630..2648)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    2922..3043
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2959..2978)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3013..3032
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3050..3505)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3187..3208)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3399..3418
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3532..3636
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3638..3995
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3638..3829
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3661..3681)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3846..3981
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4030..4824
                     /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             4030..4824
                     /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(4084..4103)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4694..4713
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5012..5031)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5056..5103
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5432..6020
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5921..5940
                     /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 gacttaattt ggttatggcc aaggaccgct tacaacttct
     1201 agagaagatg caaccagttt tgccattttc caagtcacaa acggacgtct ataatgacag
     1261 tactaacttg gcatgccgca atggacatct ccagtcagaa agtggagctg ttccaaaaag
     1321 aaaagacccc ttaacacaca ctagtaattc actgcctcgt tcaaaaacag ttatgaaaac
     1381 tggatctgca ggcctttcag gccaccatag agcacctagt tacagtggtt tatccatggt
     1441 ttctggagtg aaacagggat ctggtcctgc tcctaccact cataagggta ctccgaaaac
     1501 aaataggaca aataaacctt ctacccctac aactgctact cgtaagaaaa aagacttgaa
     1561 gaattttagg aatgtggaca gcaaccttgc taaccttata atgaatgaaa ttgtggacaa
     1621 tggaacagct gttaaatttg atgatatagc tggtcaagac ttggcaaaac aagcattgca
     1681 agaaattgtt attcttcctt ctctgaggcc tgagttgttc acagggctta gagctcctgc
     1741 cagagggctg ttactctttg gtccacctgg gaatgggaag acaatgctgg ctaaagcagt
     1801 agctgcagaa tcgaatgcaa ccttctttaa tataagtgct gcaagtttaa cttcaaaata
     1861 cgtgggagaa ggagagaaat tggtgagggc tctttttgct gtggctcgag aacttcaacc
     1921 ttctataatt tttatagatg aagttgatag ccttttgtgt gaaagaagag aaggggagca
     1981 cgatgctagt agacgcctaa aaactgaatt tctaatagaa tttgatggtg tacagtctgc
     2041 tggagatgac agagtacttg taatgggtgc aactaatagg ccacaagagc ttgatgaggc
     2101 tgttctcagg cgtttcatca aacgggtata tgtgtcttta ccaaatgagg agacaagact
     2161 acttttgctt aaaaatctgt tatgtaaaca aggaagtcca ttgacccaaa aagaactagc
     2221 acaacttgct agaatgactg atggatactc aggaagtgac ctaacagctt tggcaaaaga
     2281 tgcagcactg ggtcctatcc gagaactaaa accagaacag gtgaagaata tgtctgccag
     2341 tgagatgaga aatattcgat tatctgactt cactgaatcc ttgaaaaaaa taaaacgcag
     2401 cgtcagccct caaactttag aagcgtacat acgttggaac aaggactttg gagataccac
     2461 tgtttaagga aatacctttg taaacctgca gaacatttta cttaaaagag gaaacacaag
     2521 atcccccggg ctgcaggaat tcgatatcaa gcttatcgat accgtcgacc tcgagggggg
     2581 gcccggtacc ttaattaatt aaggtaccag gtaagtgtac ccaattcgcc ctatagtgag
     2641 tcgtattaca attcactcga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
     2701 agatccaatt tttaagtgta taatgtgtta aactactgat tctaattgtt tgtgtatttt
     2761 agattcacag tcccaaggct catttcaggc ccctcagtcc tcacagtctg ttcatgatca
     2821 taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc
     2881 ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt
     2941 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     3001 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaacgcgta aattgtaagc
     3061 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     3121 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
     3181 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
     3241 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
     3301 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
     3361 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
     3421 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
     3481 cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     3541 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     3601 tgataaatgc ttcaataata ttgaaaaagg aagaatcctg aggcggaaag aaccagctgt
     3661 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     3721 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     3781 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     3841 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     3901 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
     3961 gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac aggatgagga
     4021 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     4081 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     4141 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     4201 aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     4261 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     4321 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     4381 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     4441 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     4501 ctggacgaag aacatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgagc
     4561 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     4621 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     4681 tatcaggaca tagcgttggc tacccgtgat attgctgaag aacttggcgg cgaatgggct
     4741 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     4801 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     4861 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     4921 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     4981 agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca ataccggaag
     5041 gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt gggtcgtttg
     5101 ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc
     5161 attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg
     5221 aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc tcaggttact
     5281 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     5341 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     5401 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     5461 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     5521 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     5581 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     5641 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     5701 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     5761 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     5821 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     5881 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     5941 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     6001 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     6061 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     6121 ttaccgccat gcattagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca
     6181 tatatggagt tccg
//
