LOCUS       sequence-477715-        8044 bp    DNA     circular SYN 03-JUL-2025
DEFINITION  Expresses WT rat NF186 with a C-terminal HA tag in mammalian cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8044)
  AUTHORS   Stajkovic N, Liu Y, Arsic A, Meng N, Lyu H, Zhang N, Grimm D, Lerche
            H, Nikic-Spiegel I
  TITLE     Direct fluorescent labeling of NF186 and NaV1.6 in living primary 
            neurons using bioorthogonal click chemistry.
  JOURNAL   J Cell Sci. 2023 Jun 15;136(12):jcs260600. doi: 10.1242/jcs.260600. 
            Epub 2023 Jun 28.
  PUBMED    37288813
REFERENCE   2  (bases 1 to 8044)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 3, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Cell 
            Sci."; date: "2023-06-15"; volume: "136(12):jcs260600. doi"; pages: 
            " 10.1242/jcs.260600. Epub 2023 Jun 28"
COMMENT     Sequence Label: CMV-NF186WT-HA
FEATURES             Location/Qualifiers
     source          1..8044
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        100..402
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        403..606
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     557..577
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             4719..4745
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    4870..4991
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4907..4926)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4961..4980
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4998..5453)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5135..5156)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5347..5366
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5480..5584
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5586..5943
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5586..5777
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5609..5629)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5794..5929
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5978..6772
                     /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             5978..6772
                     /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(6032..6051)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6642..6661
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6960..6979)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7004..7051
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7380..7968
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7869..7888
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cccctgattc tgtggataac cgtattaccg ccatgcatta gttattaata gtaatcaatt
       61 acggggtcat tagttcatag cccatatatg gagttccgtg ttacataact tacggtaaat
      121 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
      181 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
      241 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
      301 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
      361 acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag
      421 tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt
      481 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac
      541 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc
      601 agagctggtt tagtgaaccg tcagatccgc tagcgctacc ggactcagat ctcgagctca
      661 agcttgatat cgaattccgg gcggcggctg gcggcgagag ctgctgcagg ggacgcgggc
      721 gaagtggcgg cgccgggagc agacatctgg gactgcgccc agggctggag ctggagctcc
      781 tggaggtaag gttgactgac ttatgtgcga tttgggagcc tggagttttc ttgccctctg
      841 ccgctgccgt caaccggagc agagcctccg ctggtgttgc gaaaagcagg ccgaatgagg
      901 cagagaagct gagcactgtc tggaaagccc acgaggaacg gcgcaaggca tcgagagccc
      961 gtgggctggg cagtgcggag cagggcctgg cgctgaggat ggccaggcag caggcgccac
     1021 cctgggtcca cgtagccctc atcctcttcc tcctcagcct cggaggggcc attgagattc
     1081 cgatggatcc aagcattcag aatgagctga cccaaccccc aacgatcacc aagcagtcgg
     1141 tgaaggacca catcgtggac ccccgagata acatcctgat tgaatgtgaa gctaaaggga
     1201 accccgcccc cagtttccac tggactcgca acagcaggtt cttcaacatt gccaaggacc
     1261 cacgggtgtc catgaggagg aggtctggga ccttggtgat cgacttccgc agtggtgggc
     1321 ggcctgagga gtacgaaggg gagtaccagt gctttgcccg gaacaaattc ggcacagctc
     1381 ttagcaaccg catccgcctg caggtgtcca aatctcccct gtggcccaag gaaaacctag
     1441 accccgtcgt ggttcaagag ggtgccccct taaccctgca gtgcaacccc ccacctggcc
     1501 tcccatcccc cgtcatcttc tggatgagca gctccatgga gcccatcacc caggacaagc
     1561 gtgtctccca gggtcacaac ggggacctgt acttctccaa cgtcatgctg caggacatgc
     1621 agaccgacta cagctgcaat gcacgcttcc acttcaccca caccattcag cagaagaatc
     1681 ccttcaccct caaagtcctc accacccgag gagttgcgga aagaacacct agcttcatgt
     1741 atccccaggg cacgtcaagc agtcagatgg tactgcgcgg catggacctg ctgctggagt
     1801 gcattgcctc tggggtccca acaccagata ttgcatggta caagaaaggt ggggacctcc
     1861 catctgacaa ggccaagttc gagaatttta acaaggctct gcgcatcacc aatgtctctg
     1921 aagaggactc tggggagtat ttctgcctgg cctccaacaa gatgggcagc atccggcaca
     1981 cgatctcggt gagagtaaag gctgccccat actggctgga tgagcccaag aacttgatcc
     2041 tggctcctgg tgaagatggg aggctggtgt gtcgagccaa tgggaacccg aagccgaccg
     2101 tccagtggct ggtgaatgga gaccctttgc aatcggcacc acccaacccc aaccgtgagg
     2161 tggccggaga cactatcatc ttccgggaca ctcagatcag cagcagggca gtgtaccagt
     2221 gtaacacatc caacgaacat ggctacctgc tggccaatgc cttcgtcagc gtattagatg
     2281 taccccctcg gatgctgtct ccccggaacc agctcatcag ggtgatcctt tacaaccgga
     2341 cgcgactgga ctgtccgttc tttgggtctc ccattccaac actccgatgg tttaagaatg
     2401 ggcaaggaag caacctggat ggtggtaact accacgtcta cgaaaatggc agcctggaaa
     2461 tcaagatgat tcgcaaagag gaccaaggca tctacacctg tgtggccacc aacatcctgg
     2521 gaaaagctga aaatcaagtc cgcctggagg tcaaagaccc caccaggatc tacaggatgc
     2581 ctgaagacca ggtggccaag aggggcacca cagtgcagct ggagtgccgt gtgaagcatg
     2641 acccctcctt gaaactcaca gtctcctggc tgaaggatga cgagccactc tacattggaa
     2701 acaggatgaa aaaggaagat gactccctga ccatcttcgg agtggcagag cgggaccaag
     2761 gcagttacac gtgcatggcc agcaccgagc tggaccagga cctggcaaag gcctacctca
     2821 ctgttctagc tgatcaggcc actccgacta accgtttggc tgccctgccc aaagggcggc
     2881 cagaccgacc cagggacctg gagctcactg acctggcgga aaggagtgtg aggctgacct
     2941 ggatcccggg ggatgataac aacagtccca tcacagacta cgttgtccag tttgaagagg
     3001 accagttcca gccaggagtc tggcatgacc actccaagtt cccaggcagt gtcaactcag
     3061 ccgtcctcca tctgtccccg tatgtcaact atcagttccg agtcatcgct gtcaacgagg
     3121 ttgggagcag ccaccccagc cttccatccg agcggtaccg aacaagcggg gcaccccctg
     3181 aatccaaccc cagtgatgtg aagggcgaag ggacacgaaa aaacaatatg gagatcacat
     3241 ggacgcctat gaatgctacc tccgcctttg gccccaacct gcgctacatt gtcaagtggc
     3301 gacggagaga aacccgagag acttggaaca atgtcaccgt gtggggctct cgctatgtgg
     3361 tggggcagac ccctgtctac gtaccctatg agatccgagt ccaggctgaa aatgactttg
     3421 ggaaaggccc ggagcctgaa accgtcattg ggtactcggg ggaagattta cccagtgccc
     3481 ccaggcgttt cagagtccga cagcccaacc tggagaccat caacctggaa tgggatcacc
     3541 cagagcaccc caacgggatc ctgattggat acacgctcag atacgtgccc tttaatggaa
     3601 ccaaactggg aaagcagatg gtggaaaact tctctcccaa tcagaccaag ttctcggtgc
     3661 agagagcaga ccccgtgtcc cgttaccgct tctccctcag tgccaggacg caggtgggct
     3721 ctggagaagc agccacggag gagtccccaa cacctccaaa tgaagctact ccaactgcag
     3781 ctcctcccac attgcccccg actaccgtgg gtaccacagg ccttgtgagc agtactgatg
     3841 ctactgccct tgctgccacc agtgaagcca caacagttcc catcatccca accgtcgtac
     3901 ctaccaccgt cgccaccacc attgccacaa ctactacaac cactgccgcc gccaccacca
     3961 ctaccaccac ggagagtcct cccactacca ccactgggac taagattcac gaaaccgccc
     4021 ccgacgagca gtccatttgg aacgtcacgg tgctccccaa cagtaaatgg gccaacatca
     4081 cctggaagca caatttcagg cctggaactg actttgtggt tgagtacatc gacagcaacc
     4141 atacgaaaaa aactgtccct gttaaggccc aagcccagcc catacagctg acagacctct
     4201 ttcccgggat gacgtacacg ttgcgggtgt attcccggga caacgagggc atcagcagta
     4261 ccgtcatcac ctttatgacc agtacagctt acaccaacaa ccagactgac atcgccaccc
     4321 agggctggtt catcgggctc atgtgtgcca ttgcccttct ggtgctgatc ctgcttatcg
     4381 tctgcttcat caagaggagt cgtggcggca agtacccagt gcgagaaaag aaggatgtcc
     4441 ccttgggccc tgaagacccc aaagaagaag atggttcatt tgactacagt gacgaggaca
     4501 acaagcccct gcagggcagc cagacatctc tggacggcac catcaagcag caggagagtg
     4561 acgacagcct agtggactac ggcgagggtg gcgagggcca gttcaacgaa gatggctcct
     4621 ttattggcca gtacactgtc agaaaggaca aggaggagac cgagggcaat gagagctcag
     4681 aggccacatc tccagtcaat gccatctatt ccctggccta cccatacgac gtcccagact
     4741 acgcttgagc ggccgcgact ctagatcata atcagccata ccacatttgt agaggtttta
     4801 cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt
     4861 gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     4921 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     4981 aatgtatctt aaggcgtaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt
     5041 ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc
     5101 aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt
     5161 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact
     5221 acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg
     5281 gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag
     5341 aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac
     5401 gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg
     5461 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     5521 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     5581 gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     5641 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     5701 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     5761 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     5821 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     5881 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     5941 aaagatcgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
     6001 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     6061 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     6121 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc
     6181 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     6241 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     6301 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     6361 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     6421 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     6481 cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg tcgtgaccca
     6541 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     6601 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     6661 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     6721 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     6781 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     6841 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     6901 atcctccagc gcggggatct catgctggag ttcttcgccc accctagggg gaggctaact
     6961 gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag
     7021 aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg
     7081 cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt
     7141 tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg
     7201 gcaggccctg ccatagcctc aggttactca tatatacttt agattgattt aaaacttcat
     7261 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     7321 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     7381 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     7441 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     7501 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     7561 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     7621 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     7681 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     7741 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     7801 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     7861 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     7921 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     7981 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     8041 ttat
//
