LOCUS       sequence-477518-        8044 bp    DNA     circular SYN 01-JUL-2025
DEFINITION  Expresses rat NF186 with a TAG codon at position 534 and a 
            C-terminal HA tag in mammalian cells. Can be used for click 
            labeling..
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 1, 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-NF186(K534TAG)-HA
FEATURES             Location/Qualifiers
     source          1..8044
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        95..398
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        399..602
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     553..573
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             4715..4741
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    4866..4987
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4903..4922)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4957..4976
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4994..5449)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5131..5152)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5343..5362
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5476..5580
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5582..5939
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5582..5773
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5605..5625)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5790..5925
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5974..6768
                     /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             5974..6768
                     /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(6028..6047)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6638..6657
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6956..6975)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7000..7047
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7376..7964
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7865..7884
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tgattctgtg gataaccgta ttaccgccat gcattagtta ttaatagtaa tcaattacgg
       61 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
      121 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
      181 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
      241 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
      301 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
      361 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca
      421 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
      481 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
      541 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
      601 ctggtttagt gaaccgtcag atccgctagc gctaccggac tcagatctcg agctcaagct
      661 tgatatcgaa ttccgggcgg cggctggcgg cgagagctgc tgcaggggac gcgggcgaag
      721 tggcggcgcc gggagcagac atctgggact gcgcccaggg ctggagctgg agctcctgga
      781 ggtaaggttg actgacttat gtgcgatttg ggagcctgga gttttcttgc cctctgccgc
      841 tgccgtcaac cggagcagag cctccgctgg tgttgcgaaa agcaggccga atgaggcaga
      901 gaagctgagc actgtctgga aagcccacga ggaacggcgc aaggcatcga gagcccgtgg
      961 gctgggcagt gcggagcagg gcctggcgct gaggatggcc aggcagcagg cgccaccctg
     1021 ggtccacgta gccctcatcc tcttcctcct cagcctcgga ggggccattg agattccgat
     1081 ggatccaagc attcagaatg agctgaccca acccccaacg atcaccaagc agtcggtgaa
     1141 ggaccacatc gtggaccccc gagataacat cctgattgaa tgtgaagcta aagggaaccc
     1201 cgcccccagt ttccactgga ctcgcaacag caggttcttc aacattgcca aggacccacg
     1261 ggtgtccatg aggaggaggt ctgggacctt ggtgatcgac ttccgcagtg gtgggcggcc
     1321 tgaggagtac gaaggggagt accagtgctt tgcccggaac aaattcggca cagctcttag
     1381 caaccgcatc cgcctgcagg tgtccaaatc tcccctgtgg cccaaggaaa acctagaccc
     1441 cgtcgtggtt caagagggtg cccccttaac cctgcagtgc aaccccccac ctggcctccc
     1501 atcccccgtc atcttctgga tgagcagctc catggagccc atcacccagg acaagcgtgt
     1561 ctcccagggt cacaacgggg acctgtactt ctccaacgtc atgctgcagg acatgcagac
     1621 cgactacagc tgcaatgcac gcttccactt cacccacacc attcagcaga agaatccctt
     1681 caccctcaaa gtcctcacca cccgaggagt tgcggaaaga acacctagct tcatgtatcc
     1741 ccagggcacg tcaagcagtc agatggtact gcgcggcatg gacctgctgc tggagtgcat
     1801 tgcctctggg gtcccaacac cagatattgc atggtacaag aaaggtgggg acctcccatc
     1861 tgacaaggcc aagttcgaga attttaacaa ggctctgcgc atcaccaatg tctctgaaga
     1921 ggactctggg gagtatttct gcctggcctc caacaagatg ggcagcatcc ggcacacgat
     1981 ctcggtgaga gtaaaggctg ccccatactg gctggatgag cccaagaact tgatcctggc
     2041 tcctggtgaa gatgggaggc tggtgtgtcg agccaatggg aacccgaagc cgaccgtcca
     2101 gtggctggtg aatggagacc ctttgcaatc ggcaccaccc aaccccaacc gtgaggtggc
     2161 cggagacact atcatcttcc gggacactca gatcagcagc agggcagtgt accagtgtaa
     2221 cacatccaac gaacatggct acctgctggc caatgccttc gtcagcgtat tagatgtacc
     2281 ccctcggatg ctgtctcccc ggaaccagct catcagggtg atcctttaca accggacgcg
     2341 actggactgt ccgttctttg ggtctcccat tccaacactc cgatggttta agaatgggca
     2401 aggaagcaac ctggatggtg gtaactacca cgtctacgaa aatggcagcc tggaaatcaa
     2461 gatgattcgc aaagaggacc aaggcatcta cacctgtgtg gccaccaaca tcctgggaaa
     2521 agctgaaaat caagtccgcc tggaggtcaa agaccccacc aggatctaca ggatgcctga
     2581 agaccaggtg gcctagaggg gcaccacagt gcagctggag tgccgtgtga agcatgaccc
     2641 ctccttgaaa ctcacagtct cctggctgaa ggatgacgag ccactctaca ttggaaacag
     2701 gatgaaaaag gaagatgact ccctgaccat cttcggagtg gcagagcggg accaaggcag
     2761 ttacacgtgc atggccagca ccgagctgga ccaggacctg gcaaaggcct acctcactgt
     2821 tctagctgat caggccactc cgactaaccg tttggctgcc ctgcccaaag ggcggccaga
     2881 ccgacccagg gacctggagc tcactgacct ggcggaaagg agtgtgaggc tgacctggat
     2941 cccgggggat gataacaaca gtcccatcac agactacgtt gtccagtttg aagaggacca
     3001 gttccagcca ggagtctggc atgaccactc caagttccca ggcagtgtca actcagccgt
     3061 cctccatctg tccccgtatg tcaactatca gttccgagtc atcgctgtca acgaggttgg
     3121 gagcagccac cccagccttc catccgagcg gtaccgaaca agcggggcac cccctgaatc
     3181 caaccccagt gatgtgaagg gcgaagggac acgaaaaaac aatatggaga tcacatggac
     3241 gcctatgaat gctacctccg cctttggccc caacctgcgc tacattgtca agtggcgacg
     3301 gagagaaacc cgagagactt ggaacaatgt caccgtgtgg ggctctcgct atgtggtggg
     3361 gcagacccct gtctacgtac cctatgagat ccgagtccag gctgaaaatg actttgggaa
     3421 aggcccggag cctgaaaccg tcattgggta ctcgggggaa gatttaccca gtgcccccag
     3481 gcgtttcaga gtccgacagc ccaacctgga gaccatcaac ctggaatggg atcacccaga
     3541 gcaccccaac gggatcctga ttggatacac gctcagatac gtgcccttta atggaaccaa
     3601 actgggaaag cagatggtgg aaaacttctc tcccaatcag accaagttct cggtgcagag
     3661 agcagacccc gtgtcccgtt accgcttctc cctcagtgcc aggacgcagg tgggctctgg
     3721 agaagcagcc acggaggagt ccccaacacc tccaaatgaa gctactccaa ctgcagctcc
     3781 tcccacattg cccccgacta ccgtgggtac cacaggcctt gtgagcagta ctgatgctac
     3841 tgcccttgct gccaccagtg aagccacaac agttcccatc atcccaaccg tcgtacctac
     3901 caccgtcgcc accaccattg ccacaactac tacaaccact gccgccgcca ccaccactac
     3961 caccacggag agtcctccca ctaccaccac tgggactaag attcacgaaa ccgcccccga
     4021 cgagcagtcc atttggaacg tcacggtgct ccccaacagt aaatgggcca acatcacctg
     4081 gaagcacaat ttcaggcctg gaactgactt tgtggttgag tacatcgaca gcaaccatac
     4141 gaaaaaaact gtccctgtta aggcccaagc ccagcccata cagctgacag acctctttcc
     4201 cgggatgacg tacacgttgc gggtgtattc ccgggacaac gagggcatca gcagtaccgt
     4261 catcaccttt atgaccagta cagcttacac caacaaccag actgacatcg ccacccaggg
     4321 ctggttcatc gggctcatgt gtgccattgc ccttctggtg ctgatcctgc ttatcgtctg
     4381 cttcatcaag aggagtcgtg gcggcaagta cccagtgcga gaaaagaagg atgtcccctt
     4441 gggccctgaa gaccccaaag aagaagatgg ttcatttgac tacagtgacg aggacaacaa
     4501 gcccctgcag ggcagccaga catctctgga cggcaccatc aagcagcagg agagtgacga
     4561 cagcctagtg gactacggcg agggtggcga gggccagttc aacgaagatg gctcctttat
     4621 tggccagtac actgtcagaa aggacaagga ggagaccgag ggcaatgaga gctcagaggc
     4681 cacatctcca gtcaatgcca tctattccct ggcctaccca tacgacgtcc cagactacgc
     4741 ttgagcggcc gcgactctag atcataatca gccataccac atttgtagag gttttacttg
     4801 ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg
     4861 ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     4921 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     4981 tatcttaagg cgtaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt
     5041 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     5101 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     5161 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     5221 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     5281 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     5341 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
     5401 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact
     5461 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     5521 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     5581 cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     5641 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     5701 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     5761 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     5821 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
     5881 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag
     5941 atcgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     6001 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     6061 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     6121 aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg
     6181 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     6241 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     6301 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     6361 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     6421 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     6481 ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc
     6541 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     6601 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     6661 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     6721 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     6781 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     6841 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     6901 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa
     6961 cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata
     7021 aaacgcacgg tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact
     7081 ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc
     7141 caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag
     7201 gccctgccat agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt
     7261 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     7321 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     7381 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     7441 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     7501 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     7561 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     7621 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     7681 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     7741 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     7801 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     7861 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     7921 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     7981 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     8041 cccc
//
