LOCUS       sequence-260968-        7190 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human NFM.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7190)
  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 7190)
  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: phNFM
FEATURES             Location/Qualifiers
     source          1..7190
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        79..382
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        383..586
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     537..557
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     complement(3824..3840)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3825..3841)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    3996..4117
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4033..4052)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4087..4106
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4124..4579)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4261..4282)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4473..4492
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4606..4710
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4712..5069
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4712..4903
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4735..4755)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4920..5055
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5104..5898
                     /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             5104..5898
                     /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(5158..5177)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5768..5787
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6086..6105)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6130..6177
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6506..7094
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6995..7014
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgtattaccg ccatgcatta gttattaata gtaatcaatt acggggtcat tagttcatag
       61 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
      121 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
      181 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
      241 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
      301 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
      361 attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata
      421 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
      481 ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca
      541 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctggtt tagtgaaccg
      601 tcagatccgc tagcgctacc ggactcagat ctcgagctca agcttcgaat tcgccaccat
      661 gagctacacg ttggactcgc tgggcaaccc gtccgcctac cggcgggtaa ccgagacccg
      721 ctcgagcttc agccgcgtca gcggctcccc gtccagtggc ttccgctcgc agtcgtggtc
      781 ccgcggctcg cccagcaccg tgtcctcctc ctataagcgc agcatgctcg ccccgcgcct
      841 cgcttacagc tcggccatgc tcagctccgc cgagagcagc cttgacttca gccagtcctc
      901 gtccctgctc aacggcggct ccggacccgg cggcgactac aagctgtccc gctccaacga
      961 gaaggagcag ctgcaggggc tgaacgaccg ctttgccggc tacatagaga aggtgcacta
     1021 cctggagcag cagaataagg agattgaggc ggagatccag gcgctgcggc agaagcaggc
     1081 ctcgcacgcc cagctgggcg acgcgtacga ccaggagatc cgcgagctgc gcgccaccct
     1141 ggagatggtg aaccacgaga aggctcaggt gcagctggac tcggaccacc tggaggaaga
     1201 catccaccgg ctcaaggagc gctttgagga ggaggcgcgg ttgcgcgacg acactgaggc
     1261 ggccatccgc gcgctgcgca aagacatcga ggaggcgtcg ctggtcaagg tggagctgga
     1321 caagaaggtg cagtcgctgc aggatgaggt ggccttcctg cggagcaacc acgaggagga
     1381 ggtggccgac cttctggccc agatccaggc atcgcacatc acggtggagc gcaaagacta
     1441 cctgaagaca gacatctcga cggcgctgaa ggaaatccgc tcccagctcg aaagccactc
     1501 agaccagaat atgcaccagg ccgaagagtg gttcaaatgc cgctacgcca agctcaccga
     1561 ggcggccgag cagaacaagg aggccatccg ctccgccaag gaagagatcg ccgagtaccg
     1621 gcgccagctg cagtccaaga gcatcgagct agagtcggtg cgcggcacca aggagtccct
     1681 ggagcggcag ctcagcgaca tcgaggagcg ccacaaccac gacctcagca gctaccagga
     1741 caccatccag cagctggaaa atgagcttcg gggcacaaag tgggaaatgg ctcgtcattt
     1801 gcgcgaatac caggacctcc tcaacgtcaa gatggctctg gatatagaaa tcgctgcgta
     1861 cagaaaactc ctggagggtg aagagactag atttagcaca tttgcaggaa gcatcactgg
     1921 gccactgtat acacaccgac ccccaatcac aatatccagt aagattcaga aacccaaggt
     1981 ggaagctccc aagcttaagg tccaacacaa atttgtcgag gagatcatag aggaaaccaa
     2041 agtggaggat gagaagtcag aaatggaaga ggccctgaca gccattacag aggaattggc
     2101 cgtttccatg aaggaagaga agaaagaagc agcagaagaa aaggaagagg aacccgaagc
     2161 tgaagaagaa gaagtagctg ccaaaaagtc tccagtgaaa gcaactgcac ctgaagttaa
     2221 agaagaggaa ggggaaaagg aggaagaaga aggccaggaa gaagaggagg aagaagatga
     2281 gggagctaag tcagaccaag ccgaagaggg aggatccgag aaggaaggct ctagtgaaaa
     2341 agaggaaggt gagcaggaag aaggagaaac agaagctgaa gctgaaggag aggaagccga
     2401 agctaaagag gaaaagaaag tggaggaaaa gagtgaggaa gtggctacca aggaggagct
     2461 ggtggcagat gccaaggtgg aaaagccaga aaaagccaag tctcctgtgc caaaatcacc
     2521 agtggaagag aaaggcaagt ctcctgtgcc caagtcacca gtggaagaga aaggcaagtc
     2581 tcctgtgccc aagtcaccag tggaagagaa aggcaagtct cctgtgccga aatcaccagt
     2641 ggaagagaaa ggcaagtctc ctgtgtcaaa atcaccagtg gaagagaaag ccaaatctcc
     2701 tgtgccaaaa tcaccagtgg aagaggcaaa gtcaaaagca gaagtgggga aaggtgaaca
     2761 gaaagaggaa gaagaaaagg aagtcaagga agctcccaag gaagagaagg tagagaaaaa
     2821 ggaagagaaa ccaaaggatg tgccagagaa gaagaaagct gagtcccctg taaaggagga
     2881 agctgtggca gaggtggtca ccatcaccaa atcggtaaag gtgcacttgg agaaagagac
     2941 caaagaagag gggaagccac tgcagcagga gaaagagaag gagaaagcgg gaggagaggg
     3001 aggaagtgag gaggaaggga gtgataaagg tgccaaggga tccaggaagg aagacatagc
     3061 tgtcaatggg gaggtagaag gaaaagagga ggtagagcag gagaccaagg aaaaaggcag
     3121 tgggagggaa gaggagaaag gcgttgtcac caatggccta gacttgagcc cagcagatga
     3181 aaagaagggg ggtgataaaa gtgaggagaa agtggtggtg accaaaacgg tagaaaaaat
     3241 caccagtgag gggggagatg gtgctaccaa atacatcact aaatctgtaa ccgtcactca
     3301 aaaggttgaa gagcatgaag agacctttga ggagaaacta gtgtctacta aaaaggtaga
     3361 aaaagtcact tcacacgcca tagtaaagga agtcacccag agtgactaag atttgagtcc
     3421 attgcaaaag gttaagccat atgacaattt caaaatgcat gtgattggca gcttcaaaac
     3481 agaacgggtt ctcccatggg ggctccagac attgtatttt actttgtgca atatgagggg
     3541 actgcatgca agctcagggt gctccctcct cagtctttgg gggattcaaa tgcatgatat
     3601 tgtatgtacc tgggaaattt gccgatttcc taagctgttg gaagggggtc acttaagggg
     3661 ggatgtcttg agatgtatta tgcaaagtac caactgagcc aaaaacaata aatgaaacac
     3721 agaactcagc cttaagaaag ctatatatga ataattatgt ttacctcact ggtgcattta
     3781 aaatggactt ttgttcatgg gagaacctcg ttatcaagct tatcgatacc gtcgacctcg
     3841 agggggggcc cggtaccgcg ggcccgggat ccaccggatc tagataactg atcataatca
     3901 gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga
     3961 acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg
     4021 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     4081 ctagttgtgg tttgtccaaa ctcatcaatg tatcttaacg cgtaaattgt aagcgttaat
     4141 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     4201 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     4261 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     4321 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     4381 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     4441 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     4501 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     4561 gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     4621 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     4681 atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg
     4741 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     4801 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     4861 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     4921 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     4981 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     5041 gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt
     5101 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     5161 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     5221 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     5281 ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     5341 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     5401 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     5461 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     5521 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     5581 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc
     5641 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     5701 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     5761 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     5821 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     5881 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     5941 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     6001 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     6061 tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc
     6121 gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata
     6181 aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg
     6241 gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc
     6301 cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata
     6361 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     6421 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     6481 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     6541 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     6601 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     6661 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     6721 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     6781 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     6841 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     6901 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     6961 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     7021 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     7081 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     7141 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
//
