LOCUS       sequence-357913-        5509 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For mouse Nav1.6 knockin with 3xHA at the C-terminal.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5509)
  AUTHORS   Liu H, Wang HG, Pitt GS, Liu ZJ
  TITLE     Direct Observation of Compartment-Specific Localization and Dynamics
            of Voltage-Gated Sodium Channels.
  JOURNAL   J Neurosci. 2022 Jun 6;42(28):5482-98. doi: 
            10.1523/JNEUROSCI.0086-22.2022.
  PUBMED    35672149
REFERENCE   2  (bases 1 to 5509)
  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"; doi: 
            "10.1523/JNEUROSCI.0086-22"; journalName: "J Neurosci"; date: 
            "2022-06-6- 6"; volume: "42"; issue: "28"; pages: "5482-98"
COMMENT     Sequence Label: PX552-mScn8a-3xHA-Syn-smFP-flag
FEATURES             Location/Qualifiers
     source          1..5509
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        145..393
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     145..165
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     316..335
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        414..489
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             544..570
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             586..612
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YPYDVPDYA"
     CDS             586..612
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             628..654
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        720..1167
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      1181..1190
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1190..1213
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1217..1240
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1244..1267
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1796..1819
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1826..1849
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1862..1885
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1892..1915
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2126..2149
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2153..2176
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2180..2203
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    2256..2732
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2455..2474)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   2772..2912
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2772..2901
                     /label=AAV2 ITR
     rep_origin      2987..3442
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3074..3093)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3284..3305
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3459..3478)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3578..3600
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3638..3656)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3724..3828
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3829..3897,3898..4689)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4047..4066)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4860..5448
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5349..5368
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtgagggc ctatttccca tgattccttc atatttgcat
      181 atacgataca aggctgttag agagataatt ggaattaatt tgactgtaaa cacaaagata
      241 ttagtacaaa atacgtgacg tagaaagtaa taatttcttg ggtagtttgc agttttaaaa
      301 ttatgtttta aaatggacta tcatatgctt accgtaactt gaaagtattt cgatttcttg
      361 gctttatata tcttgtggaa aggacgaaac accccgacaa ggagaagcag caggttttag
      421 agctagaaat agcaagttaa aataaggcta gtccgttatc aacttgaaaa agtggcaccg
      481 agtcggtgct ttttttctag taccggtccg ctgctgcttc tccttgtcgg ttctggatcg
      541 ggttacccat acgatgttcc agattacgct ggcagtggta gcggttaccc ctacgacgtg
      601 cccgactacg ccggctccgg cagtggttat ccttatgacg tccctgatta cgccggctca
      661 ggtagcggcc cgctgctgct tctccttgtc ggtctagact gcagagggcc ctgcgtatga
      721 gtgcaagtgg gttttaggac caggatgagg cggggtgggg gtgcctacct gacgaccgac
      781 cccgacccac tggacaagca cccaaccccc attccccaaa ttgcgcatcc cctatcagag
      841 agggggaggg gaaacaggat gcggcgaggc gcgtgcgcac tgccagcttc agcaccgcgg
      901 acagtgcctt cgcccccgcc tggcggcgcg cgccaccgcc gcctcagcac tgaaggcgcg
      961 ctgacgtcac tcgccggtcc cccgcaaact ccccttcccg gccaccttgg tcgcgtccgc
     1021 gccgccgccg gcccagccgg accgcaccac gcgaggcgcg agataggggg gcacgggcgc
     1081 gaccatctgc gctgcggcgc cggcgactca gcgctgcctc agtctgcggt gggcagcgga
     1141 ggagtcgtgt cgtgcctgag agcgcagtcg agaaggtacc gccaccatgg actacaagga
     1201 cgacgacgat aaaggagatt acaaagatga cgatgataag ggcgattata aggacgacga
     1261 tgacaagggc ggcgtgtcga aaggcgagga gctgttcaca ggcgtggtcc ccatcctggt
     1321 tgagctggac ggtgacgtta acggccacaa atttagcgta cgtggtgagg gcgagggtga
     1381 cgctaccaac ggaaagctga ccctgaaatt catctgcaca actggcaagc tgccggtgcc
     1441 ctggcccacg ctggtaacca ctctgggcgg aggcgttcaa tgtttcagtc gttacccgga
     1501 tcacatgaag cagcatgact tcttcaagtc ggccatgccc gaaggctacg tgcaggagcg
     1561 caccatcagt tttaaggatg acggcaccta caagactcgc gccgaggtca agttcgaagg
     1621 agacaccctg gttaatcgta tcgagctcaa aggcatcgat ttcaaggagg acggaaatat
     1681 tctgggccac aagctggagt acaattttaa cagccacaat gtttacatta ctgccgacaa
     1741 gcagaagaac ggcatcaagg ctaatttcaa gattcgtcac aatgtggagg gaggcgacta
     1801 taaggatgac gatgacaaac aacaagacta taaggacgat gatgacaaag gtcagcaagg
     1861 cgattacaag gacgatgacg ataagcagca agattacaag gatgatgatg ataagggagg
     1921 cgacggaagc gtgcagctgg cagatcatta ccaacagaac acgcccattg gtgacggtcc
     1981 agtcctgctg cctgataacc actacttgtc cacccagtcg gtgctgtcga aggacccgaa
     2041 cgagaagcgt gaccacatgg tgctgctgga gttcgtgacc gccgcaggaa tcaccctcgg
     2101 catggatgag ctttacaagg gcggagatta taaagatgac gacgacaagg gcgactacaa
     2161 agatgatgat gacaagggtg actacaaaga cgacgatgat aaataagaat tcgatatcaa
     2221 gcttatcgat accgagcgct gctcgagaga tctacgggtg gcatccctgt gacccctccc
     2281 cagtgcctct cctggccctg gaagttgcca ctccagtgcc caccagcctt gtcctaataa
     2341 aattaagttg catcattttg tctgactagg tgtccttcta taatattatg gggtggaggg
     2401 gggtggtatg gagcaagggg caagttggga agacaacctg tagggcctgc ggggtctatt
     2461 gggaaccaag ctggagtgca gtggcacaat cttggctcac tgcaatctcc gcctcctggg
     2521 ttcaagcgat tctcctgcct cagcctcccg agttgttggg attccaggca tgcatgacca
     2581 ggctcagcta atttttgttt ttttggtaga gacggggttt caccatattg gccaggctgg
     2641 tctccaactc ctaatctcag gtgatctacc caccttggcc tcccaaattg ctgggattac
     2701 aggcgtgaac cactgctccc ttccctgtcc ttctgatttt gtaggtaacc acgtgcggac
     2761 cgagcggccg caggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc
     2821 gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc
     2881 agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt ttctccttac
     2941 gcatctgtgc ggtatttcac accgcatacg tcaaagcaac catagtacgc gccctgtagc
     3001 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     3061 gccttagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     3121 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     3181 ctcgacccca aaaaacttga tttgggtgat ggttcacgta gtgggccatc gccctgatag
     3241 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     3301 actggaacaa cactcaactc tatctcgggc tattcttttg atttataagg gattttgccg
     3361 atttcggtct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     3421 aaaatattaa cgtttacaat tttatggtgc actctcagta caatctgctc tgatgccgca
     3481 tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
     3541 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
     3601 ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta
     3661 taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat
     3721 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     3781 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     3841 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     3901 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     3961 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     4021 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     4081 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     4141 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     4201 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     4261 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     4321 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     4381 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     4441 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     4501 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     4561 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     4621 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     4681 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     4741 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     4801 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     4861 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     4921 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     4981 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     5041 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     5101 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     5161 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     5221 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     5281 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     5341 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     5401 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     5461 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgt
//
