LOCUS       sequence-174164-        6134 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses myc-M1S46X spastin isoform in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6134)
  AUTHORS   Solowska JM, Garbern JY, Baas PW
  TITLE     Evaluation of loss of function as an explanation for SPG4-based 
            hereditary spastic paraplegia.
  JOURNAL   Hum Mol Genet. 2010 Jul 15;19(14):2767-79. doi: 10.1093/hmg/ddq177. 
            Epub 2010 Apr 29.
  PUBMED    20430936
REFERENCE   2  (bases 1 to 6134)
  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: 
            "10.1093/hmg/ddq177"; journalName: "Hum Mol Genet"; date: 
            "2010-07-15- 15"; volume: "19"; issue: "14"; pages: "2767-79"
COMMENT     Sequence Label: pCMV-Tag 3A/myc-M1S46X
FEATURES             Location/Qualifiers
     source          1..6134
                     /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"
     regulatory      604..613
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             613..642
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(2569..2588)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2570..2588)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    2862..2983
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2899..2918)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2953..2972
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2990..3445)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3127..3148)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3339..3358
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3472..3576
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3578..3935
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3578..3769
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3601..3621)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3786..3921
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3970..4764
                     /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             3970..4764
                     /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(4024..4043)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4634..4653
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4952..4971)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4996..5043
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5372..5960
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5861..5880
                     /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 gctggagctc caccgcggtg
      601 gcggccgcca tggagcagaa actcatctct gaagaggatc tggcccgggc ggatcccccg
      661 ggctgcagga attctccggg tggacgaggg aagaagaaag gctccggcgg cgccagcaac
      721 ccggtgcctc ccaggcctcc gcccccttgc ctggcccccg cccctcccgc cgccgggccg
      781 cgcccctccg cccgagtcgc cgcataagcg gaacctgtac tatttctcct acccgctgtt
      841 tgtaggcttc gcgctgctgc gtttggtcgc cttccacctg gggctcctct tcgtgtggct
      901 ctgccagcgc ttctcccgcg ccctcatggc agccaagagg agctccgggg ccgcgccagc
      961 acctgcctcg gcctcggccc cggcgccggt gccgggcggc gaggccgagc gcgtccgagt
     1021 cttccacaaa caggccttcg agtacatctc cattgccctg cgcatcgatg aggatgagaa
     1081 agcaggacag aaggagcaag ctgtggaatg gtataagaaa ggtattgaag aactggaaaa
     1141 aggaatagct gttatagtta caggacaagg tgaacagtgt gaaagagcta gacgccttca
     1201 agctaaaatg atgactaatt tggttatggc caaggaccgc ttacaacttc tagagaagat
     1261 gcaaccagtt ttgccatttt ccaagtcaca aacggacgtc tataatgaca gtactaactt
     1321 ggcatgccgc aatggacatc tccagtcaga aagtggagct gttccaaaaa gaaaagaccc
     1381 cttaacacac actagtaatt cactgcctcg ttcaaaaaca gttatgaaaa ctggatctgc
     1441 aggcctttca ggccaccata gagcacctag ttacagtggt ttatccatgg tttctggagt
     1501 gaaacaggga tctggtcctg ctcctaccac tcataagggt actccgaaaa caaataggac
     1561 aaataaacct tctaccccta caactgctac tcgtaagaaa aaagacttga agaattttag
     1621 gaatgtggac agcaaccttg ctaaccttat aatgaatgaa attgtggaca atggaacagc
     1681 tgttaaattt gatgatatag ctggtcaaga cttggcaaaa caagcattgc aagaaattgt
     1741 tattcttcct tctctgaggc ctgagttgtt cacagggctt agagctcctg ccagagggct
     1801 gttactcttt ggtccacctg ggaatgggaa gacaatgctg gctaaagcag tagctgcaga
     1861 atcgaatgca accttcttta atataagtgc tgcaagttta acttcaaaat acgtgggaga
     1921 aggagagaaa ttggtgaggg ctctttttgc tgtggctcga gaacttcaac cttctataat
     1981 ttttatagat gaagttgata gccttttgtg tgaaagaaga gaaggggagc acgatgctag
     2041 tagacgccta aaaactgaat ttctaataga atttgatggt gtacagtctg ctggagatga
     2101 cagagtactt gtaatgggtg caactaatag gccacaagag cttgatgagg ctgttctcag
     2161 gcgtttcatc aaacgggtat atgtgtcttt accaaatgag gagacaagac tacttttgct
     2221 taaaaatctg ttatgtaaac aaggaagtcc attgacccaa aaagaactag cacaacttgc
     2281 tagaatgact gatggatact caggaagtga cctaacagct ttggcaaaag atgcagcact
     2341 gggtcctatc cgagaactaa aaccagaaca ggtgaagaat atgtctgcca gtgagatgag
     2401 aaatattcga ttatctgact tcactgaatc cttgaaaaaa ataaaacgca gcgtcagccc
     2461 tcaaacttta gaagcgtaca tacgttggaa caaggacttt ggagatacca ctgtttaagg
     2521 gcccggtacc ttaattaatt aaggtaccag gtaagtgtac ccaattcgcc ctatagtgag
     2581 tcgtattaca attcactcga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
     2641 agatccaatt tttaagtgta taatgtgtta aactactgat tctaattgtt tgtgtatttt
     2701 agattcacag tcccaaggct catttcaggc ccctcagtcc tcacagtctg ttcatgatca
     2761 taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc
     2821 ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt
     2881 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     2941 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaacgcgta aattgtaagc
     3001 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     3061 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
     3121 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
     3181 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
     3241 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
     3301 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
     3361 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
     3421 cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     3481 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     3541 tgataaatgc ttcaataata ttgaaaaagg aagaatcctg aggcggaaag aaccagctgt
     3601 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     3661 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     3721 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     3781 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     3841 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
     3901 gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac aggatgagga
     3961 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     4021 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     4081 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     4141 aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     4201 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     4261 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     4321 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     4381 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     4441 ctggacgaag aacatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgagc
     4501 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     4561 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     4621 tatcaggaca tagcgttggc tacccgtgat attgctgaag aacttggcgg cgaatgggct
     4681 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     4741 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     4801 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     4861 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     4921 agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca ataccggaag
     4981 gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt gggtcgtttg
     5041 ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc
     5101 attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg
     5161 aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc tcaggttact
     5221 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     5281 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     5341 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     5401 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     5461 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     5521 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     5581 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     5641 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     5701 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     5761 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     5821 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     5881 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     5941 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     6001 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     6061 ttaccgccat gcattagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca
     6121 tatatggagt tccg
//
