LOCUS       sequence-260965-        6063 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human NFL containing the Charcot-Marie-Tooth disease type 
            2E L268P mutation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6063)
  AUTHORS   Stone EJ, Uchida A, Brown A
  TITLE     Charcot-Marie-Tooth disease Type 2E/1F mutant neurofilament proteins
            assemble into neurofilaments.
  JOURNAL   Cytoskeleton (Hoboken). 2019 Jul;76(7-8):423-439. doi: 
            10.1002/cm.21566. Epub 2019 Nov 6.
  PUBMED    31574566
REFERENCE   2  (bases 1 to 6063)
  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: 
            "Cytoskeleton (Hoboken)."; date: "2019-07"; volume: 
            "76(7-8):423-439. doi"; pages: " 10.1002/cm.21566. Epub 2019 Nov 6"
COMMENT     Sequence Label: phNFL-L268P
FEATURES             Location/Qualifiers
     source          1..6063
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        41..344
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        345..548
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     499..519
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    589..645
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    2831..2952
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2868..2887)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2922..2941
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2959..3414)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3096..3117)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3308..3327
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3441..3545
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3547..3904
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3547..3738
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3570..3590)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3755..3890
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3939..4733
                     /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             3939..4733
                     /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(3993..4012)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4603..4622
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4921..4940)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4965..5012
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5341..5929
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5830..5849
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
       61 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
      121 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
      181 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
      241 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
      301 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
      361 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
      421 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
      481 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
      541 gcagagctgg tttagtgaac cgtcagatcc gctagcgcta ccggactcag atctcgagct
      601 caagcttcga attctgcagt cgacggtacc gcgggcccgg gatccgggtg gcagaacggc
      661 gccgagccgc acacagccat ccatcctccc ccttccctct ctcccctgtc ctctctctcc
      721 gggctcccac cgccgccgcg ggccggggag ccaccggccg ccaccatgag ttccttcagc
      781 tacgagccgt actactcgac ctcctacaag cggcgctacg tggagacgcc ccgggtgcac
      841 atctccagcg tgcgcagcgg ctacagcacc gcacgctcag cttactccag ctactcggcg
      901 ccggtgtctt cctcgctgtc cgtgcgccgc agctactcct ccagctctgg atcgttgatg
      961 cccagtctgg agaacctcga cctgagccag gtagccgcca tcagcaacga cctcaagtcc
     1021 atccgcacgc aggagaaggc gcagctccag gacctcaatg accgcttcgc cagcttcatc
     1081 gagcgcgtgc acgagctgga gcagcagaac aaggtcctgg aagccgagct gctggtgctg
     1141 cgccagaagc actccgagcc atcccgcttc cgggcgctgt acgagcagga gatccgcgac
     1201 ctgcgcctgg cggcggaaga tgccaccaac gagaagcagg cgctccaggg cgagcgcgaa
     1261 gggctggagg agaccctgcg caacctgcag gcgcgctatg aagaggaggt gctgagccgc
     1321 gaggacgccg agggccggct gatggaagcg cgcaaaggcg ccgacgaggc ggcgctcgct
     1381 cgcgccgagc tcgagaagcg catcgacagc ttgatggacg aaatctcttt tctgaagaaa
     1441 gtgcacgaag aggagatcgc cgaactgcag gcgcagatcc agtacgcgca gatctccgtg
     1501 gagatggacg tgaccaagcc cgacctttcc gccgcgctca aggacatccg cgcgcagtac
     1561 gagaagccgg ccgccaagaa catgcagaac gctgaggaat ggttcaagag ccgcttcacc
     1621 gtgctgaccg agagcgccgc caagaacacc gacgccgtgc gcgccgccaa ggacgaggtg
     1681 tccgagagcc gtcgtctgct caaggccaag accctggaaa tcgaagcatg ccggggcatg
     1741 aatgaagcgc tggagaagca gctgcaggag ctggaggaca agcagaacgc cgacatcagc
     1801 gctatgcagg acacgatcaa caaattagaa aatgaattga ggaccacaaa gagtgaaatg
     1861 gcacgatacc taaaagaata ccaagacctc ctcaacgtga agatggcttt ggatattgag
     1921 attgcagctt acaggaaact cttggaaggc gaggagaccc gactcagttt caccagcgtg
     1981 ggaagcataa ccagtggcta ctcccagagc tcccaggtct ttggccgatc tgcctacggc
     2041 ggtttacaga ccagctccta tctgatgtcc acccgctcct tcccgtccta ctacaccagc
     2101 catgtccaag aggagcagat cgaagtggag gaaaccattg aggctgccaa ggctgaggaa
     2161 gccaaggatg agcccccctc tgaaggagaa gccgaggagg aggagaagga caaggaagag
     2221 gccgaggaag aggaggcagc tgaagaggaa gaagctgcca aggaagagtc tgaagaagca
     2281 aaagaagaag aagaaggagg tgaaggtgaa gaaggagagg aaaccaaaga agctgaagag
     2341 gaggagaaga aagttgaagg tgctggggag gaacaagcag ctaagaagaa agattgaacc
     2401 cccatttcct taattatttc aggaataatt ctcccgaaat caggtcaacc ccatcaccaa
     2461 ccaaccaacc agttgagttc cagattctat gtgaattaaa aagtcaatat atgtataatt
     2521 ctgagatgac ttaggttgga cattcaatgt tgtgctatga atttcctctt tatgcagagt
     2581 atctgtttgc ttgcagagtg gctttctggc ttgctgccag cctgtgcatg gtccacaagc
     2641 cgaattcaga tctggtacca cgcgtatcga taagcttgca tgcctgcagg tcgactctag
     2701 aggatccacc ggatctagat aactgatcat aatcagccat accacatttg tagaggtttt
     2761 acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat
     2821 tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     2881 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     2941 caatgtatct taacgcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
     3001 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     3061 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
     3121 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     3181 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     3241 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     3301 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     3361 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg
     3421 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     3481 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     3541 agagtcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc
     3601 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     3661 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     3721 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     3781 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     3841 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     3901 caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     3961 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     4021 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     4081 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat
     4141 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     4201 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     4261 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     4321 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     4381 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     4441 ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc
     4501 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     4561 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     4621 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     4681 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     4741 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     4801 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     4861 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac
     4921 tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca
     4981 gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg
     5041 gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt
     5101 ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc
     5161 ggcaggccct gccatagcct caggttactc atatatactt tagattgatt taaaacttca
     5221 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     5281 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     5341 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     5401 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     5461 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     5521 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     5581 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     5641 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     5701 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     5761 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     5821 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     5881 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     5941 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     6001 gttatcccct gattctgtgg ataaccgtat taccgccatg cattagttat taatagtaat
     6061 caa
//
