LOCUS       sequence-357904-        5697 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For mouse Nav1.2 knockin with 3xV5 at the C-terminal.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5697)
  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 5697)
  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-mScn2a-3xV5-EF1-smFP-flag
FEATURES             Location/Qualifiers
     source          1..5697
                     /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             546..587
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             603..644
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             660..701
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     promoter        857..1068
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             1081..1349
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     regulatory      1369..1378
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1378..1401
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1405..1428
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1432..1455
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1984..2007
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2014..2037
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2050..2073
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2080..2103
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2314..2337
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2341..2364
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2368..2391
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    2444..2920
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2643..2662)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   2960..3100
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2960..3089
                     /label=AAV2 ITR
     rep_origin      3175..3630
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3262..3281)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3472..3493
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3647..3666)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3766..3788
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3826..3844)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3912..4016
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4017..4085,4086..4877)
                     /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(4235..4254)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5048..5636
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5537..5556
                     /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 accggacaag gggaaagata tcagttttag
      421 agctagaaat agcaagttaa aataaggcta gtccgttatc aacttgaaaa agtggcaccg
      481 agtcggtgct ttttttctag taccggtccc tgatatcttt ccccttgtcc ggttctggat
      541 cgggtgggaa gcctataccc aatcctctcc tggggttgga cagcaccggc agtggtagcg
      601 gtggcaagcc catccccaac cccctgctgg gcctggactc caccggctcc ggcagtggtg
      661 gaaaacctat tcccaaccca ctgctgggac tggattctac aggctcaggt agcggctccc
      721 tgatatcttt ccccttgtcc tctagaccaa ggatctgcga tcgctccggt gcccgtcagt
      781 gggcagagcg cacatcgccc acagtccccg agaagttggg gggaggggtc ggcaattcgc
      841 tccggtgccc gtcagtgggc agagcgcaca tcgcccacag tccccgagaa gttgggggga
      901 ggggtcggca attgaacggg tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat
      961 gtcgtgtact ggctccgcct ttttcccgag ggtgggggag aaccgtatat aagtgcagta
     1021 gtcgccgtga acgttctttt tcgcaacggg tttgccgcca gaacacagct gaagcttcga
     1081 ggggctcgca tctctccttc acgcgcccgc cgccctacct gaggccgcca tccacgccgg
     1141 ttgagtcgcg ttctgccgcc tcccgcctgt ggtgcctcct gaactgcgtc cgccgtctag
     1201 gtaagtttaa agctcaggtc gagaccgggc ctttgtccgg cgctcccttg gagcctacct
     1261 agactcagcc ggctctccac gctttgcctg accctgcttg ctcaactcta cgtctttgtt
     1321 tcgttttctg ttctgcgccg ttacagatcc aagctgtgac cgggtaccgc caccatggac
     1381 tacaaggacg acgacgataa aggagattac aaagatgacg atgataaggg cgattataag
     1441 gacgacgatg acaagggcgg cgtgtcgaaa ggcgaggagc tgttcacagg cgtggtcccc
     1501 atcctggttg agctggacgg tgacgttaac ggccacaaat ttagcgtacg tggtgagggc
     1561 gagggtgacg ctaccaacgg aaagctgacc ctgaaattca tctgcacaac tggcaagctg
     1621 ccggtgccct ggcccacgct ggtaaccact ctgggcggag gcgttcaatg tttcagtcgt
     1681 tacccggatc acatgaagca gcatgacttc ttcaagtcgg ccatgcccga aggctacgtg
     1741 caggagcgca ccatcagttt taaggatgac ggcacctaca agactcgcgc cgaggtcaag
     1801 ttcgaaggag acaccctggt taatcgtatc gagctcaaag gcatcgattt caaggaggac
     1861 ggaaatattc tgggccacaa gctggagtac aattttaaca gccacaatgt ttacattact
     1921 gccgacaagc agaagaacgg catcaaggct aatttcaaga ttcgtcacaa tgtggaggga
     1981 ggcgactata aggatgacga tgacaaacaa caagactata aggacgatga tgacaaaggt
     2041 cagcaaggcg attacaagga cgatgacgat aagcagcaag attacaagga tgatgatgat
     2101 aagggaggcg acggaagcgt gcagctggca gatcattacc aacagaacac gcccattggt
     2161 gacggtccag tcctgctgcc tgataaccac tacttgtcca cccagtcggt gctgtcgaag
     2221 gacccgaacg agaagcgtga ccacatggtg ctgctggagt tcgtgaccgc cgcaggaatc
     2281 accctcggca tggatgagct ttacaagggc ggagattata aagatgacga cgacaagggc
     2341 gactacaaag atgatgatga caagggtgac tacaaagacg acgatgataa ataagaattc
     2401 gatatcaagc ttatcgatac cgagcgctgc tcgagagatc tacgggtggc atccctgtga
     2461 cccctcccca gtgcctctcc tggccctgga agttgccact ccagtgccca ccagccttgt
     2521 cctaataaaa ttaagttgca tcattttgtc tgactaggtg tccttctata atattatggg
     2581 gtggaggggg gtggtatgga gcaaggggca agttgggaag acaacctgta gggcctgcgg
     2641 ggtctattgg gaaccaagct ggagtgcagt ggcacaatct tggctcactg caatctccgc
     2701 ctcctgggtt caagcgattc tcctgcctca gcctcccgag ttgttgggat tccaggcatg
     2761 catgaccagg ctcagctaat ttttgttttt ttggtagaga cggggtttca ccatattggc
     2821 caggctggtc tccaactcct aatctcaggt gatctaccca ccttggcctc ccaaattgct
     2881 gggattacag gcgtgaacca ctgctccctt ccctgtcctt ctgattttgt aggtaaccac
     2941 gtgcggaccg agcggccgca ggaaccccta gtgatggagt tggccactcc ctctctgcgc
     3001 gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg
     3061 gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt
     3121 ctccttacgc atctgtgcgg tatttcacac cgcatacgtc aaagcaacca tagtacgcgc
     3181 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     3241 ttgccagcgc cttagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     3301 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     3361 tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc
     3421 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     3481 tgttccaaac tggaacaaca ctcaactcta tctcgggcta ttcttttgat ttataaggga
     3541 ttttgccgat ttcggtctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     3601 attttaacaa aatattaacg tttacaattt tatggtgcac tctcagtaca atctgctctg
     3661 atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     3721 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     3781 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc
     3841 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
     3901 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     3961 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     4021 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     4081 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     4141 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     4201 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     4261 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     4321 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     4381 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     4441 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     4501 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     4561 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     4621 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     4681 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     4741 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     4801 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     4861 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     4921 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     4981 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     5041 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     5101 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     5161 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     5221 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     5281 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     5341 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     5401 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     5461 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     5521 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     5581 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     5641 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgt
//
