LOCUS       sequence-363086-        5636 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses wild-type mouse neurofilament light chain with a 
            C-terminal FLAG tag (DYKDDDDK) in mammalian cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5636)
  AUTHORS   Arsic A, Hagemann C, Stajkovic N, Schubert T, Nikic-Spiegel I
  TITLE     Minimal genetically encoded tags for fluorescent protein labeling in
            living neurons.
  JOURNAL   Nat Commun. 2022 Jan 14;13(1):314. doi: 10.1038/s41467-022-27956-y.
  PUBMED    35031604
REFERENCE   2  (bases 1 to 5636)
  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: "Nat 
            Commun."; date: "2022-01-14"; volume: "13(1):314. doi"; pages: " 
            10.1038/s41467-022-27956-y"
COMMENT     Sequence Label: pCMV_NFL(WT)-FLAG
FEATURES             Location/Qualifiers
     source          1..5636
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1685..1708
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    1833..1954
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1870..1889)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1924..1943
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1961..2416)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2098..2119)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2310..2329
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2443..2547
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2549..2906
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2549..2740
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2572..2592)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2757..2892
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2941..3735
                     /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             2941..3735
                     /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(2995..3014)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3605..3624
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3923..3942)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3967..4014
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4343..4931
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4832..4851
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        5106..5409
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5410..5613
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5564..5584
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 ctagcgctac cggactcaga tctcgagctc aagcttcacc atgagttcgt tcggctacga
       61 tccgtacttt tcgacctcct acaagcggcg ctatgtggag acgccccggg tgcacatctc
      121 cagcgtgcgc agcggctaca gcacggcgcg ctccgcgtac tccagctact ccgcgccggt
      181 ctcctcctcg ctgtccgtgc gccgcagcta ctcgtccagc tctggctctt tgatgcccag
      241 cctggagaat ctcgatctga gccaggtagc cgccatcagc aacgacctca agtctatccg
      301 cacacaagag aaggcacagc tgcaggacct caacgatcgc ttcgccagct tcatcgagcg
      361 cgtgcacgag ctggagcagc agaacaaggt cctggaagcc gagctgttgg tgctgcgcca
      421 gaaacactct gagccttccc gcttccgcgc cctgtacgag caggagatcc gcgatctgcg
      481 gctggcagcg gaagacgcca ctaacgagaa gcaggcgctg cagggcgagc gcgaggggct
      541 ggaggagact ctgcgcaacc tgcaggctcg ctatgaggaa gaagtgctga gccgcgagga
      601 cgccgagggc cggctgatgg aagcgcgcaa aggtgcggat gaggccgcgc tcgcccgcgc
      661 cgagctggag aagcgcatcg acagcctgat ggacgagata gctttcctga agaaggtgca
      721 cgaggaagag atcgccgagc tgcaggctca gatccagtat gctcagatct ccgtggagat
      781 ggacgtgtcc tccaagcccg acctctccgc cgctctcaag gacatccgcg ctcagtacga
      841 gaagctggcc gccaagaaca tgcagaacgc cgaagagtgg ttcaagagcc gcttcaccgt
      901 gctaaccgag agcgccgcca agaacaccga cgctgtgcgc gctgccaagg acgaggtgtc
      961 ggaaagccgc cgcctgctca aggctaagac cctggagatc gaagcctgcc ggggtatgaa
     1021 cgaagctctg gagaagcagc tgcaggagct agaggacaag cagaatgcag acattagcgc
     1081 catgcaggac acaatcaaca aactggagaa tgagctgaga agcacgaaga gcgagatggc
     1141 caggtacctg aaggagtacc aggacctcct caatgtcaag atggccttgg acatcgagat
     1201 tgcagcttac agaaaactct tggaaggcga agagaccagg ctcagtttca ccagcgtggg
     1261 tagcataacc agcggctact ctcagagctc gcaggtcttc ggccgttctg cttacagtgg
     1321 cttgcagagc agctcctact tgatgtctgc tcgctctttc ccagcctact ataccagcca
     1381 cgtccaggaa gagcagacag aggtcgagga gaccattgag gctacgaaag ctgaggaggc
     1441 caaggatgag cccccctctg aaggagaagc agaagaggag gagaaggaga aagaggaggg
     1501 agaggaagag gaaggcgctg aggaggaaga agctgccaag gatgagtctg aagacacaaa
     1561 agaagaagaa gaaggtggtg agggtgagga ggaagacacc aaagaatctg aagaggaaga
     1621 gaagaaagag gagagtgctg gagaggagca ggtggctaag aagaaagatg gggcccggga
     1681 tccggactac aaagacgatg acgacaagtg agcggccgcg actctagatc ataatcagcc
     1741 ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc
     1801 tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt
     1861 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     1921 gttgtggttt gtccaaactc atcaatgtat cttaaggcgt aaattgtaag cgttaatatt
     1981 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
     2041 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca
     2101 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
     2161 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg
     2221 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
     2281 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg
     2341 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
     2401 ccgctacagg gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     2461 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     2521 cttcaataat attgaaaaag gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt
     2581 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     2641 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     2701 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     2761 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     2821 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     2881 tttttggagg cctaggcttt tgcaaagatc gatcaagaga caggatgagg atcgtttcgc
     2941 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     3001 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     3061 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     3121 caagacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     3181 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     3241 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     3301 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     3361 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     3421 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac
     3481 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     3541 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     3601 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     3661 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     3721 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
     3781 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     3841 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     3901 cccaccctag ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg
     3961 ctatgacggc aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt gttcataaac
     4021 gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc
     4081 aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag
     4141 ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc ctcaggttac tcatatatac
     4201 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     4261 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     4321 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     4381 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     4441 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     4501 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     4561 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     4621 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     4681 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     4741 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     4801 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     4861 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     4921 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     4981 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcca
     5041 tgcattagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
     5101 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
     5161 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
     5221 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
     5281 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
     5341 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
     5401 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
     5461 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
     5521 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
     5581 cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca gatccg
//
