LOCUS       sequence-173570-        6133 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses myc-M1L195V spastin isoform in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6133)
  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 6133)
  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-M1L195V
FEATURES             Location/Qualifiers
     source          1..6133
                     /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(2568..2587)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2569..2587)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    2861..2982
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2898..2917)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2952..2971
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2989..3444)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3126..3147)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3338..3357
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3471..3575
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3577..3934
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3577..3768
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3600..3620)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3785..3920
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3969..4763
                     /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             3969..4763
                     /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(4023..4042)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4633..4652
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4951..4970)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4995..5042
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5371..5959
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5860..5879
                     /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 gcccctccgc ccgagtcgcc gcataagcgg aacctgtact atttctccta cccgctgttt
      841 gtaggcttcg cgctgctgcg tttggtcgcc ttccacctgg ggctcctctt cgtgtggctc
      901 tgccagcgct tctcccgcgc cctcatggca gccaagagga gctccggggc cgcgccagca
      961 cctgcctcgg cctcggcccc ggcgccggtg ccgggcggcg aggccgagcg cgtccgagtc
     1021 ttccacaaac aggccttcga gtacatctcc attgccctgc gcatcgatga ggatgagaaa
     1081 gcaggacaga aggagcaagc tgtggaatgg tataagaaag gtattgaaga actggaaaaa
     1141 ggaatagctg ttatagttac aggacaaggt gaacagtgtg aaagagctag acgccttcaa
     1201 gctaaaatga tgactaattt ggttatggcc aaggaccgct tacaacttgt agagaagatg
     1261 caaccagttt tgccattttc caagtcacaa acggacgtct ataatgacag tactaacttg
     1321 gcatgccgca atggacatct ccagtcagaa agtggagctg ttccaaaaag aaaagacccc
     1381 ttaacacaca ctagtaattc actgcctcgt tcaaaaacag ttatgaaaac tggatctgca
     1441 ggcctttcag gccaccatag agcacctagt tacagtggtt tatccatggt ttctggagtg
     1501 aaacagggat ctggtcctgc tcctaccact cataagggta ctccgaaaac aaataggaca
     1561 aataaacctt ctacccctac aactgctact cgtaagaaaa aagacttgaa gaattttagg
     1621 aatgtggaca gcaaccttgc taaccttata atgaatgaaa ttgtggacaa tggaacagct
     1681 gttaaatttg atgatatagc tggtcaagac ttggcaaaac aagcattgca agaaattgtt
     1741 attcttcctt ctctgaggcc tgagttgttc acagggctta gagctcctgc cagagggctg
     1801 ttactctttg gtccacctgg gaatgggaag acaatgctgg ctaaagcagt agctgcagaa
     1861 tcgaatgcaa ccttctttaa tataagtgct gcaagtttaa cttcaaaata cgtgggagaa
     1921 ggagagaaat tggtgagggc tctttttgct gtggctcgag aacttcaacc ttctataatt
     1981 tttatagatg aagttgatag ccttttgtgt gaaagaagag aaggggagca cgatgctagt
     2041 agacgcctaa aaactgaatt tctaatagaa tttgatggtg tacagtctgc tggagatgac
     2101 agagtacttg taatgggtgc aactaatagg ccacaagagc ttgatgaggc tgttctcagg
     2161 cgtttcatca aacgggtata tgtgtcttta ccaaatgagg agacaagact acttttgctt
     2221 aaaaatctgt tatgtaaaca aggaagtcca ttgacccaaa aagaactagc acaacttgct
     2281 agaatgactg atggatactc aggaagtgac ctaacagctt tggcaaaaga tgcagcactg
     2341 ggtcctatcc gagaactaaa accagaacag gtgaagaata tgtctgccag tgagatgaga
     2401 aatattcgat tatctgactt cactgaatcc ttgaaaaaaa taaaacgcag cgtcagccct
     2461 caaactttag aagcgtacat acgttggaac aaggactttg gagataccac tgtttaaggg
     2521 cccggtacct taattaatta aggtaccagg taagtgtacc caattcgccc tatagtgagt
     2581 cgtattacaa ttcactcgat cgcccttccc aacagttgcg cagcctgaat ggcgaatgga
     2641 gatccaattt ttaagtgtat aatgtgttaa actactgatt ctaattgttt gtgtatttta
     2701 gattcacagt cccaaggctc atttcaggcc cctcagtcct cacagtctgt tcatgatcat
     2761 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc
     2821 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
     2881 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     2941 gcattctagt tgtggtttgt ccaaactcat caatgtatct taacgcgtaa attgtaagcg
     3001 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     3061 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     3121 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     3181 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
     3241 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     3301 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     3361 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     3421 ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     3481 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     3541 gataaatgct tcaataatat tgaaaaagga agaatcctga ggcggaaaga accagctgtg
     3601 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3661 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     3721 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     3781 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     3841 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
     3901 aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat
     3961 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     4021 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     4081 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     4141 atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     4201 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     4261 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     4321 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     4381 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     4441 tggacgaaga acatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca
     4501 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     4561 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     4621 atcaggacat agcgttggct acccgtgata ttgctgaaga acttggcggc gaatgggctg
     4681 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     4741 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac
     4801 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     4861 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     4921 gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg
     4981 aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt
     5041 tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca
     5101 ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga
     5161 aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc
     5221 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     5281 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     5341 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     5401 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     5461 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     5521 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     5581 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     5641 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     5701 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     5761 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     5821 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     5881 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     5941 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     6001 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     6061 taccgccatg cattagttat taatagtaat caattacggg gtcattagtt catagcccat
     6121 atatggagtt ccg
//
