LOCUS       sequence-173597-        6083 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses S46X spastin M1 and M87 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6083)
  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 6083)
  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/S46X without 5'UTR
FEATURES             Location/Qualifiers
     source          1..6083
                     /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(2518..2537)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2519..2537)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    2811..2932
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2848..2867)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2902..2921
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2939..3394)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3076..3097)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3288..3307
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3421..3525
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3527..3884
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3527..3718
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3550..3570)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3735..3870
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3919..4713
                     /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             3919..4713
                     /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(3973..3992)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4583..4602
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4901..4920)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4945..4992
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5321..5909
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5810..5829
                     /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 cagtgagagc tgtgaatgaa ttctccgggt ggacgaggga agaagaaagg ctccggcggc
      661 gccagcaacc cggtgcctcc caggcctccg cccccttgcc tggcccccgc ccctcccgcc
      721 gccgggccgc gcccctccgc ccgagtcgcc gcataagcgg aacctgtact atttctccta
      781 cccgctgttt gtaggcttcg cgctgctgcg tttggtcgcc ttccacctgg ggctcctctt
      841 cgtgtggctc tgccagcgct tctcccgcgc cctcatggca gccaagagga gctccggggc
      901 cgcgccagca cctgcctcgg cctcggcccc ggcgccggtg ccgggcggcg aggccgagcg
      961 cgtccgagtc ttccacaaac aggccttcga gtacatctcc attgccctgc gcatcgatga
     1021 ggatgagaaa gcaggacaga aggagcaagc tgtggaatgg tataagaaag gtattgaaga
     1081 actggaaaaa ggaatagctg ttatagttac aggacaaggt gaacagtgtg aaagagctag
     1141 acgccttcaa gctaaaatga tgactaattt 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 tgtttaaggg cccggtacct taattaatta aggtaccagg taagtgtacc caattcgccc
     2521 tatagtgagt cgtattacaa ttcactcgat cgcccttccc aacagttgcg cagcctgaat
     2581 ggcgaatgga gatccaattt ttaagtgtat aatgtgttaa actactgatt ctaattgttt
     2641 gtgtatttta gattcacagt cccaaggctc atttcaggcc cctcagtcct cacagtctgt
     2701 tcatgatcat aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc
     2761 cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta
     2821 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     2881 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct taacgcgtaa
     2941 attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
     3001 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
     3061 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
     3121 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa
     3181 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
     3241 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
     3301 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
     3361 cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg
     3421 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     3481 caataaccct gataaatgct tcaataatat tgaaaaagga agaatcctga ggcggaaaga
     3541 accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     3601 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     3661 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     3721 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     3781 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     3841 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca
     3901 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     3961 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     4021 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     4081 ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc
     4141 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     4201 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     4261 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     4321 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     4381 caggatgatc tggacgaaga acatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     4441 aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     4501 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     4561 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga acttggcggc
     4621 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     4681 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     4741 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     4801 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     4861 tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa
     4921 taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg
     4981 ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac
     5041 cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa
     5101 gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct
     5161 caggttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     5221 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     5281 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     5341 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     5401 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     5461 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     5521 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     5581 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     5641 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     5701 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     5761 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     5821 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     5881 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     5941 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     6001 ataaccgtat taccgccatg cattagttat taatagtaat caattacggg gtcattagtt
     6061 catagcccat atatggagtt ccg
//
