LOCUS       sequence-363050-        5621 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mouse neurofilament light chain with a TAG codon at the 
            position R438 and an N-terminal FLAG tag (DYKDDDDK) in mammalian 
            cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5621)
  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 5621)
  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_FLAG-NFL(R438TAG)
FEATURES             Location/Qualifiers
     source          1..5621
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             44..67
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    1818..1939
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1855..1874)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1909..1928
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1946..2401)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2083..2104)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2295..2314
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2428..2532
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2534..2891
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2534..2725
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2557..2577)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2742..2877
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2926..3720
                     /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             2926..3720
                     /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(2980..2999)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3590..3609
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3908..3927)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3952..3999
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4328..4916
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4817..4836
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        5091..5394
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5395..5598
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5549..5569
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 ctagcgctac cggactcaga tctcgagctc aagcttcacc atggactaca aagacgatga
       61 cgacaagagt tcgttcggct acgatccgta cttttcgacc tcctacaagc ggcgctatgt
      121 ggagacgccc cgggtgcaca tctccagcgt gcgcagcggc tacagcacgg cgcgctccgc
      181 gtactccagc tactccgcgc cggtctcctc ctcgctgtcc gtgcgccgca gctactcgtc
      241 cagctctggc tctttgatgc ccagcctgga gaatctcgat ctgagccagg tagccgccat
      301 cagcaacgac ctcaagtcta tccgcacaca agagaaggca cagctgcagg acctcaacga
      361 tcgcttcgcc agcttcatcg agcgcgtgca cgagctggag cagcagaaca aggtcctgga
      421 agccgagctg ttggtgctgc gccagaaaca ctctgagcct tcccgcttcc gcgccctgta
      481 cgagcaggag atccgcgatc tgcggctggc agcggaagac gccactaacg agaagcaggc
      541 gctgcagggc gagcgcgagg ggctggagga gactctgcgc aacctgcagg ctcgctatga
      601 ggaagaagtg ctgagccgcg aggacgccga gggccggctg atggaagcgc gcaaaggtgc
      661 ggatgaggcc gcgctcgccc gcgccgagct ggagaagcgc atcgacagcc tgatggacga
      721 gatagctttc ctgaagaagg tgcacgagga agagatcgcc gagctgcagg ctcagatcca
      781 gtatgctcag atctccgtgg agatggacgt gtcctccaag cccgacctct ccgccgctct
      841 caaggacatc cgcgctcagt acgagaagct ggccgccaag aacatgcaga acgccgaaga
      901 gtggttcaag agccgcttca ccgtgctaac cgagagcgcc gccaagaaca ccgacgctgt
      961 gcgcgctgcc aaggacgagg tgtcggaaag ccgccgcctg ctcaaggcta agaccctgga
     1021 gatcgaagcc tgccggggta tgaacgaagc tctggagaag cagctgcagg agctagagga
     1081 caagcagaat gcagacatta gcgccatgca ggacacaatc aacaaactgg agaatgagct
     1141 gagaagcacg aagagcgaga tggccaggta cctgaaggag taccaggacc tcctcaatgt
     1201 caagatggcc ttggacatcg agattgcagc ttacagaaaa ctcttggaag gcgaagagac
     1261 caggctcagt ttcaccagcg tgggtagcat aaccagcggc tactctcaga gctcgcaggt
     1321 cttcggccgt tctgcttaca gtggcttgca gagcagctcc tacttgatgt ctgcttagtc
     1381 tttcccagcc tactatacca gccacgtcca ggaagagcag acagaggtcg aggagaccat
     1441 tgaggctacg aaagctgagg aggccaagga tgagcccccc tctgaaggag aagcagaaga
     1501 ggaggagaag gagaaagagg agggagagga agaggaaggc gctgaggagg aagaagctgc
     1561 caaggatgag tctgaagaca caaaagaaga agaagaaggt ggtgagggtg aggaggaaga
     1621 caccaaagaa tctgaagagg aagagaagaa agaggagagt gctggagagg agcaggtggc
     1681 taagaagaaa gattgagcgg ccgcgactct agatcataat cagccatacc acatttgtag
     1741 aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga
     1801 atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     1861 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     1921 aactcatcaa tgtatcttaa ggcgtaaatt gtaagcgtta atattttgtt aaaattcgcg
     1981 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     2041 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     2101 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     2161 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     2221 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     2281 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     2341 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     2401 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     2461 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     2521 aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg
     2581 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     2641 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     2701 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     2761 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     2821 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     2881 gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     2941 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     3001 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     3061 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg
     3121 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     3181 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     3241 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     3301 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     3361 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     3421 gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc
     3481 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     3541 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     3601 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     3661 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     3721 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     3781 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     3841 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga
     3901 ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa
     3961 aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc
     4021 agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt
     4081 cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg
     4141 tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag attgatttaa
     4201 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     4261 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     4321 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     4381 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     4441 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     4501 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     4561 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     4621 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     4681 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     4741 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     4801 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     4861 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     4921 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     4981 ttcctgcgtt atcccctgat tctgtggata accgtattac cgccatgcat tagttattaa
     5041 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
     5101 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
     5161 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
     5221 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc
     5281 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
     5341 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg
     5401 cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt
     5461 ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca
     5521 aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag
     5581 gtctatataa gcagagctgg tttagtgaac cgtcagatcc g
//
