LOCUS       sequence-173599-        6401 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 6401)
  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 6401)
  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 with 5'UTR
FEATURES             Location/Qualifiers
     source          1..6401
                     /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(2764..2780)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2765..2781)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(2836..2855)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2837..2855)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    3129..3250
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3166..3185)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3220..3239
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3257..3712)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3394..3415)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3606..3625
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3739..3843
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3845..4202
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3845..4036
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3868..3888)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4053..4188
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4237..5031
                     /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             4237..5031
                     /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(4291..4310)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4901..4920
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5219..5238)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5263..5310
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5639..6227
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6128..6147
                     /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 gctggaggac ccacgcgtcc
      601 gggcccgagc caccgactgc aggaggagaa ggggttgtgc tcctggccga ggaaggagaa
      661 aggggcgggg ccggcgggca gcgtgcggca gtgcggagct cctgagaccg gcgggcacac
      721 gggggtctgt ggcccccgcc gtagcagtgg ctgccgccgt cgcttggttc ccgtcggtct
      781 gcgggaggcg ggttatggcg gcggcggcag tgagagctgt gaatgaattc tccgggtgga
      841 cgagggaaga agaaaggctc cggcggcgcc agcaacccgg tgcctcccag gcctccgccc
      901 ccttgcctgg cccccgcccc tcccgccgcc gggccgcgcc cctccgcccg agtcgccgca
      961 taagcggaac ctgtactatt tctcctaccc gctgtttgta ggcttcgcgc tgctgcgttt
     1021 ggtcgccttc cacctggggc tcctcttcgt gtggctctgc cagcgcttct cccgcgccct
     1081 catggcagcc aagaggagct ccggggccgc gccagcacct gcctcggcct cggccccggc
     1141 gccggtgccg ggcggcgagg ccgagcgcgt ccgagtcttc cacaaacagg ccttcgagta
     1201 catctccatt gccctgcgca tcgatgagga tgagaaagca ggacagaagg agcaagctgt
     1261 ggaatggtat aagaaaggta ttgaagaact ggaaaaagga atagctgtta tagttacagg
     1321 acaaggtgaa cagtgtgaaa gagctagacg ccttcaagct aaaatgatga ctaatttggt
     1381 tatggccaag gaccgcttac aacttctaga gaagatgcaa ccagttttgc cattttccaa
     1441 gtcacaaacg gacgtctata atgacagtac taacttggca tgccgcaatg gacatctcca
     1501 gtcagaaagt ggagctgttc caaaaagaaa agacccctta acacacacta gtaattcact
     1561 gcctcgttca aaaacagtta tgaaaactgg atctgcaggc ctttcaggcc accatagagc
     1621 acctagttac agtggtttat ccatggtttc tggagtgaaa cagggatctg gtcctgctcc
     1681 taccactcat aagggtactc cgaaaacaaa taggacaaat aaaccttcta cccctacaac
     1741 tgctactcgt aagaaaaaag acttgaagaa ttttaggaat gtggacagca accttgctaa
     1801 ccttataatg aatgaaattg tggacaatgg aacagctgtt aaatttgatg atatagctgg
     1861 tcaagacttg gcaaaacaag cattgcaaga aattgttatt cttccttctc tgaggcctga
     1921 gttgttcaca gggcttagag ctcctgccag agggctgtta ctctttggtc cacctgggaa
     1981 tgggaagaca atgctggcta aagcagtagc tgcagaatcg aatgcaacct tctttaatat
     2041 aagtgctgca agtttaactt caaaatacgt gggagaagga gagaaattgg tgagggctct
     2101 ttttgctgtg gctcgagaac ttcaaccttc tataattttt atagatgaag ttgatagcct
     2161 tttgtgtgaa agaagagaag gggagcacga tgctagtaga cgcctaaaaa ctgaatttct
     2221 aatagaattt gatggtgtac agtctgctgg agatgacaga gtacttgtaa tgggtgcaac
     2281 taataggcca caagagcttg atgaggctgt tctcaggcgt ttcatcaaac gggtatatgt
     2341 gtctttacca aatgaggaga caagactact tttgcttaaa aatctgttat gtaaacaagg
     2401 aagtccattg acccaaaaag aactagcaca acttgctaga atgactgatg gatactcagg
     2461 aagtgaccta acagctttgg caaaagatgc agcactgggt cctatccgag aactaaaacc
     2521 agaacaggtg aagaatatgt ctgccagtga gatgagaaat attcgattat ctgacttcac
     2581 tgaatccttg aaaaaaataa aacgcagcgt cagccctcaa actttagaag cgtacatacg
     2641 ttggaacaag gactttggag ataccactgt ttaaggaaat acctttgtaa acctgcagaa
     2701 cattttactt aaaagaggaa acacaagatc ccccgggctg caggaattcg atatcaagct
     2761 tatcgatacc gtcgacctcg agggggggcc cggtacctta attaattaag gtaccaggta
     2821 agtgtaccca attcgcccta tagtgagtcg tattacaatt cactcgatcg cccttcccaa
     2881 cagttgcgca gcctgaatgg cgaatggaga tccaattttt aagtgtataa tgtgttaaac
     2941 tactgattct aattgtttgt gtattttaga ttcacagtcc caaggctcat ttcaggcccc
     3001 tcagtcctca cagtctgttc atgatcataa tcagccatac cacatttgta gaggttttac
     3061 ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg
     3121 ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     3181 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     3241 atgtatctta acgcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt
     3301 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
     3361 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
     3421 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
     3481 cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
     3541 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
     3601 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
     3661 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc
     3721 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     3781 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     3841 aatcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     3901 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     3961 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     4021 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     4081 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     4141 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     4201 aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     4261 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     4321 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     4381 gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg
     4441 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     4501 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     4561 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     4621 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     4681 gaagccggtc ttgtcgatca ggatgatctg gacgaagaac atcaggggct cgcgccagcc
     4741 gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat
     4801 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     4861 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     4921 gctgaagaac ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     4981 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     5041 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     5101 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     5161 tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg aggctaactg
     5221 aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga
     5281 ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc
     5341 actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt
     5401 ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg
     5461 caggccctgc catagcctca ggttactcat atatacttta gattgattta aaacttcatt
     5521 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     5581 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     5641 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     5701 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     5761 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     5821 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     5881 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     5941 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     6001 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     6061 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     6121 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     6181 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     6241 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     6301 tatcccctga ttctgtggat aaccgtatta ccgccatgca ttagttatta atagtaatca
     6361 attacggggt cattagttca tagcccatat atggagttcc g
//
