LOCUS       sequence-260941-        6063 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human NFL.
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
FEATURES             Location/Qualifiers
     source          1..6063
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        82..385
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..589
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     540..560
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    630..686
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    2872..2993
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2909..2928)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2963..2982
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3000..3455)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3137..3158)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3349..3368
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3482..3586
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3588..3945
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3588..3779
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3611..3631)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3796..3931
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3980..4774
                     /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             3980..4774
                     /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(4034..4053)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4644..4663
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4962..4981)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5006..5053
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5382..5970
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5871..5890
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aaccgtatta ccgccatgca ttagttatta atagtaatca attacggggt cattagttca
       61 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
      121 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
      181 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
      241 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
      301 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
      361 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg
      421 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
      481 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
      541 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctg gtttagtgaa
      601 ccgtcagatc cgctagcgct accggactca gatctcgagc tcaagcttcg aattctgcag
      661 tcgacggtac cgcgggcccg ggatccgggt ggcagaacgg cgccgagccg cacacagcca
      721 tccatcctcc cccttccctc tctcccctgt cctctctctc cgggctccca ccgccgccgc
      781 gggccgggga gccaccggcc gccaccatga gttccttcag ctacgagccg tactactcga
      841 cctcctacaa gcggcgctac gtggagacgc cccgggtgca catctccagc gtgcgcagcg
      901 gctacagcac cgcacgctca gcttactcca gctactcggc gccggtgtct tcctcgctgt
      961 ccgtgcgccg cagctactcc tccagctctg gatcgttgat gcccagtctg gagaacctcg
     1021 acctgagcca ggtagccgcc atcagcaacg acctcaagtc catccgcacg caggagaagg
     1081 cgcagctcca ggacctcaat gaccgcttcg ccagcttcat cgagcgcgtg cacgagctgg
     1141 agcagcagaa caaggtcctg gaagccgagc tgctggtgct gcgccagaag cactccgagc
     1201 catcccgctt ccgggcgctg tacgagcagg agatccgcga cctgcgcctg gcggcggaag
     1261 atgccaccaa cgagaagcag gcgctccagg gcgagcgcga agggctggag gagaccctgc
     1321 gcaacctgca ggcgcgctat gaagaggagg tgctgagccg cgaggacgcc gagggccggc
     1381 tgatggaagc gcgcaaaggc gccgacgagg cggcgctcgc tcgcgccgag ctcgagaagc
     1441 gcatcgacag cttgatggac gaaatctctt ttctgaagaa agtgcacgaa gaggagatcg
     1501 ccgaactgca ggcgcagatc cagtacgcgc agatctccgt ggagatggac gtgaccaagc
     1561 ccgacctttc cgccgcgctc aaggacatcc gcgcgcagta cgagaagctg gccgccaaga
     1621 acatgcagaa cgctgaggaa tggttcaaga gccgcttcac cgtgctgacc gagagcgccg
     1681 ccaagaacac cgacgccgtg cgcgccgcca aggacgaggt gtccgagagc cgtcgtctgc
     1741 tcaaggccaa gaccctggaa atcgaagcat gccggggcat gaatgaagcg ctggagaagc
     1801 agctgcagga gctggaggac aagcagaacg ccgacatcag cgctatgcag gacacgatca
     1861 acaaattaga aaatgaattg aggaccacaa agagtgaaat ggcacgatac ctaaaagaat
     1921 accaagacct cctcaacgtg aagatggctt tggatattga gattgcagct tacaggaaac
     1981 tcttggaagg cgaggagacc cgactcagtt tcaccagcgt gggaagcata accagtggct
     2041 actcccagag ctcccaggtc tttggccgat ctgcctacgg cggtttacag accagctcct
     2101 atctgatgtc cacccgctcc ttcccgtcct actacaccag ccatgtccaa gaggagcaga
     2161 tcgaagtgga ggaaaccatt gaggctgcca aggctgagga agccaaggat gagcccccct
     2221 ctgaaggaga agccgaggag gaggagaagg acaaggaaga ggccgaggaa gaggaggcag
     2281 ctgaagagga agaagctgcc aaggaagagt ctgaagaagc aaaagaagaa gaagaaggag
     2341 gtgaaggtga agaaggagag gaaaccaaag aagctgaaga ggaggagaag aaagttgaag
     2401 gtgctgggga ggaacaagca gctaagaaga aagattgaac ccccatttcc ttaattattt
     2461 caggaataat tctcccgaaa tcaggtcaac cccatcacca accaaccaac cagttgagtt
     2521 ccagattcta tgtgaattaa aaagtcaata tatgtataat tctgagatga cttaggttgg
     2581 acattcaatg ttgtgctatg aatttcctct ttatgcagag tatctgtttg cttgcagagt
     2641 ggctttctgg cttgctgcca gcctgtgcat ggtccacaag ccgaattcag atctggtacc
     2701 acgcgtatcg ataagcttgc atgcctgcag gtcgactcta gaggatccac cggatctaga
     2761 taactgatca taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc
     2821 ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt
     2881 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     2941 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaacgcgta
     3001 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     3061 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     3121 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     3181 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
     3241 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     3301 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     3361 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     3421 acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt
     3481 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     3541 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtcctg aggcggaaag
     3601 aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     3661 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     3721 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     3781 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     3841 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     3901 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac
     3961 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc
     4021 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     4081 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     4141 cggtgccctg aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg
     4201 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     4261 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     4321 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     4381 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     4441 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     4501 caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     4561 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     4621 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     4681 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     4741 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc
     4801 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
     4861 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     4921 ctcatgctgg agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca
     4981 ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt
     5041 gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca
     5101 ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca
     5161 agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc
     5221 tcaggttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     5281 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     5341 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     5401 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     5461 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     5521 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     5581 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     5641 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     5701 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     5761 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     5821 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     5881 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     5941 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     6001 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     6061 gat
//
