LOCUS       sequence-363055-        5636 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mouse neurofilament light chain with a TAG codon at the 
            position R438 and 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(R438TAG)-FLAG
FEATURES             Location/Qualifiers
     source          1..5636
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1686..1709
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    1834..1955
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1871..1890)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1925..1944
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1962..2417)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2099..2120)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2311..2330
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2444..2548
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2550..2907
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2550..2741
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2573..2593)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2758..2893
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2942..3736
                     /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             2942..3736
                     /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(2996..3015)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3606..3625
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3924..3943)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3968..4015
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4344..4932
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4833..4852
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        5107..5410
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5411..5614
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5565..5585
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 gctagcgcta ccggactcag atctcgagct caagcttcac catgagttcg ttcggctacg
       61 atccgtactt ttcgacctcc tacaagcggc gctatgtgga gacgccccgg gtgcacatct
      121 ccagcgtgcg cagcggctac agcacggcgc gctccgcgta ctccagctac tccgcgccgg
      181 tctcctcctc gctgtccgtg cgccgcagct actcgtccag ctctggctct ttgatgccca
      241 gcctggagaa tctcgatctg agccaggtag ccgccatcag caacgacctc aagtctatcc
      301 gcacacaaga gaaggcacag ctgcaggacc tcaacgatcg cttcgccagc ttcatcgagc
      361 gcgtgcacga gctggagcag cagaacaagg tcctggaagc cgagctgttg gtgctgcgcc
      421 agaaacactc tgagccttcc cgcttccgcg ccctgtacga gcaggagatc cgcgatctgc
      481 ggctggcagc ggaagacgcc actaacgaga agcaggcgct gcagggcgag cgcgaggggc
      541 tggaggagac tctgcgcaac ctgcaggctc gctatgagga agaagtgctg agccgcgagg
      601 acgccgaggg ccggctgatg gaagcgcgca aaggtgcgga tgaggccgcg ctcgcccgcg
      661 ccgagctgga gaagcgcatc gacagcctga tggacgagat agctttcctg aagaaggtgc
      721 acgaggaaga gatcgccgag ctgcaggctc agatccagta tgctcagatc tccgtggaga
      781 tggacgtgtc ctccaagccc gacctctccg ccgctctcaa ggacatccgc gctcagtacg
      841 agaagctggc cgccaagaac atgcagaacg ccgaagagtg gttcaagagc cgcttcaccg
      901 tgctaaccga gagcgccgcc aagaacaccg acgctgtgcg cgctgccaag gacgaggtgt
      961 cggaaagccg ccgcctgctc aaggctaaga ccctggagat cgaagcctgc cggggtatga
     1021 acgaagctct ggagaagcag ctgcaggagc tagaggacaa gcagaatgca gacattagcg
     1081 ccatgcagga cacaatcaac aaactggaga atgagctgag aagcacgaag agcgagatgg
     1141 ccaggtacct gaaggagtac caggacctcc tcaatgtcaa gatggccttg gacatcgaga
     1201 ttgcagctta cagaaaactc ttggaaggcg aagagaccag gctcagtttc accagcgtgg
     1261 gtagcataac cagcggctac tctcagagct cgcaggtctt cggccgttct gcttacagtg
     1321 gcttgcagag cagctcctac ttgatgtctg cttagtcttt cccagcctac tataccagcc
     1381 acgtccagga agagcagaca gaggtcgagg agaccattga ggctacgaaa gctgaggagg
     1441 ccaaggatga gcccccctct gaaggagaag cagaagagga ggagaaggag aaagaggagg
     1501 gagaggaaga ggaaggcgct gaggaggaag aagctgccaa ggatgagtct gaagacacaa
     1561 aagaagaaga agaaggtggt gagggtgagg aggaagacac caaagaatct gaagaggaag
     1621 agaagaaaga ggagagtgct ggagaggagc aggtggctaa gaagaaagat ggggcccggg
     1681 atccggacta caaagacgat gacgacaagt gagcggccgc gactctagat cataatcagc
     1741 cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac
     1801 ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt
     1861 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     1921 agttgtggtt tgtccaaact catcaatgta tcttaaggcg taaattgtaa gcgttaatat
     1981 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     2041 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     2101 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     2161 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     2221 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     2281 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     2341 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     2401 gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     2461 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     2521 gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg
     2581 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     2641 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     2701 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     2761 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     2821 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     2881 ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg
     2941 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     3001 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     3061 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     3121 gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     3181 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     3241 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     3301 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     3361 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     3421 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga
     3481 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     3541 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     3601 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     3661 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     3721 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac
     3781 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     3841 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     3901 gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc
     3961 gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa
     4021 cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc
     4081 caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca
     4141 gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata
     4201 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     4261 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     4321 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     4381 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     4441 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     4501 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     4561 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     4621 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     4681 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     4741 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     4801 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     4861 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     4921 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     4981 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     5041 atgcattagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
     5101 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
     5161 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
     5221 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
     5281 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
     5341 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
     5401 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
     5461 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
     5521 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
     5581 gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc agatcc
//
