LOCUS       sequence-477711-        8044 bp    DNA     circular SYN 03-JUL-2025
DEFINITION  Expresses rat NF186 with a TAG codon at position 809 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 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-NF186(K809TAG)-HA
FEATURES             Location/Qualifiers
     source          1..8044
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        22..225
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     176..196
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             4338..4364
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    4489..4610
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4526..4545)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4580..4599
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4617..5072)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4754..4775)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4966..4985
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5099..5203
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5205..5562
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5205..5396
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5228..5248)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5413..5548
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5597..6391
                     /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             5597..6391
                     /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(5651..5670)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6261..6280
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6579..6598)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6623..6670
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6999..7587
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7488..7507
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        join(7763..8044,1..21)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag
       61 cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt
      121 tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa
      181 atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt
      241 cagatccgct agcgctaccg gactcagatc tcgagctcaa gcttgatatc gaattccggg
      301 cggcggctgg cggcgagagc tgctgcaggg gacgcgggcg aagtggcggc gccgggagca
      361 gacatctggg actgcgccca gggctggagc tggagctcct ggaggtaagg ttgactgact
      421 tatgtgcgat ttgggagcct ggagttttct tgccctctgc cgctgccgtc aaccggagca
      481 gagcctccgc tggtgttgcg aaaagcaggc cgaatgaggc agagaagctg agcactgtct
      541 ggaaagccca cgaggaacgg cgcaaggcat cgagagcccg tgggctgggc agtgcggagc
      601 agggcctggc gctgaggatg gccaggcagc aggcgccacc ctgggtccac gtagccctca
      661 tcctcttcct cctcagcctc ggaggggcca ttgagattcc gatggatcca agcattcaga
      721 atgagctgac ccaaccccca acgatcacca agcagtcggt gaaggaccac atcgtggacc
      781 cccgagataa catcctgatt gaatgtgaag ctaaagggaa ccccgccccc agtttccact
      841 ggactcgcaa cagcaggttc ttcaacattg ccaaggaccc acgggtgtcc atgaggagga
      901 ggtctgggac cttggtgatc gacttccgca gtggtgggcg gcctgaggag tacgaagggg
      961 agtaccagtg ctttgcccgg aacaaattcg gcacagctct tagcaaccgc atccgcctgc
     1021 aggtgtccaa atctcccctg tggcccaagg aaaacctaga ccccgtcgtg gttcaagagg
     1081 gtgccccctt aaccctgcag tgcaaccccc cacctggcct cccatccccc gtcatcttct
     1141 ggatgagcag ctccatggag cccatcaccc aggacaagcg tgtctcccag ggtcacaacg
     1201 gggacctgta cttctccaac gtcatgctgc aggacatgca gaccgactac agctgcaatg
     1261 cacgcttcca cttcacccac accattcagc agaagaatcc cttcaccctc aaagtcctca
     1321 ccacccgagg agttgcggaa agaacaccta gcttcatgta tccccagggc acgtcaagca
     1381 gtcagatggt actgcgcggc atggacctgc tgctggagtg cattgcctct ggggtcccaa
     1441 caccagatat tgcatggtac aagaaaggtg gggacctccc atctgacaag gccaagttcg
     1501 agaattttaa caaggctctg cgcatcacca atgtctctga agaggactct ggggagtatt
     1561 tctgcctggc ctccaacaag atgggcagca tccggcacac gatctcggtg agagtaaagg
     1621 ctgccccata ctggctggat gagcccaaga acttgatcct ggctcctggt gaagatggga
     1681 ggctggtgtg tcgagccaat gggaacccga agccgaccgt ccagtggctg gtgaatggag
     1741 accctttgca atcggcacca cccaacccca accgtgaggt ggccggagac actatcatct
     1801 tccgggacac tcagatcagc agcagggcag tgtaccagtg taacacatcc aacgaacatg
     1861 gctacctgct ggccaatgcc ttcgtcagcg tattagatgt accccctcgg atgctgtctc
     1921 cccggaacca gctcatcagg gtgatccttt acaaccggac gcgactggac tgtccgttct
     1981 ttgggtctcc cattccaaca ctccgatggt ttaagaatgg gcaaggaagc aacctggatg
     2041 gtggtaacta ccacgtctac gaaaatggca gcctggaaat caagatgatt cgcaaagagg
     2101 accaaggcat ctacacctgt gtggccacca acatcctggg aaaagctgaa aatcaagtcc
     2161 gcctggaggt caaagacccc accaggatct acaggatgcc tgaagaccag gtggccaaga
     2221 ggggcaccac agtgcagctg gagtgccgtg tgaagcatga cccctccttg aaactcacag
     2281 tctcctggct gaaggatgac gagccactct acattggaaa caggatgaaa aaggaagatg
     2341 actccctgac catcttcgga gtggcagagc gggaccaagg cagttacacg tgcatggcca
     2401 gcaccgagct ggaccaggac ctggcaaagg cctacctcac tgttctagct gatcaggcca
     2461 ctccgactaa ccgtttggct gccctgccca aagggcggcc agaccgaccc agggacctgg
     2521 agctcactga cctggcggaa aggagtgtga ggctgacctg gatcccgggg gatgataaca
     2581 acagtcccat cacagactac gttgtccagt ttgaagagga ccagttccag ccaggagtct
     2641 ggcatgacca ctccaagttc ccaggcagtg tcaactcagc cgtcctccat ctgtccccgt
     2701 atgtcaacta tcagttccga gtcatcgctg tcaacgaggt tgggagcagc caccccagcc
     2761 ttccatccga gcggtaccga acaagcgggg caccccctga atccaacccc agtgatgtga
     2821 agggcgaagg gacacgaaaa aacaatatgg agatcacatg gacgcctatg aatgctacct
     2881 ccgcctttgg ccccaacctg cgctacattg tcaagtggcg acggagagaa acccgagaga
     2941 cttggaacaa tgtcaccgtg tggggctctc gctatgtggt ggggcagacc cctgtctacg
     3001 taccctatga gatccgagtc caggctgaaa atgactttgg gtagggcccg gagcctgaaa
     3061 ccgtcattgg gtactcgggg gaagatttac ccagtgcccc caggcgtttc agagtccgac
     3121 agcccaacct ggagaccatc aacctggaat gggatcaccc agagcacccc aacgggatcc
     3181 tgattggata cacgctcaga tacgtgccct ttaatggaac caaactggga aagcagatgg
     3241 tggaaaactt ctctcccaat cagaccaagt tctcggtgca gagagcagac cccgtgtccc
     3301 gttaccgctt ctccctcagt gccaggacgc aggtgggctc tggagaagca gccacggagg
     3361 agtccccaac acctccaaat gaagctactc caactgcagc tcctcccaca ttgcccccga
     3421 ctaccgtggg taccacaggc cttgtgagca gtactgatgc tactgccctt gctgccacca
     3481 gtgaagccac aacagttccc atcatcccaa ccgtcgtacc taccaccgtc gccaccacca
     3541 ttgccacaac tactacaacc actgccgccg ccaccaccac taccaccacg gagagtcctc
     3601 ccactaccac cactgggact aagattcacg aaaccgcccc cgacgagcag tccatttgga
     3661 acgtcacggt gctccccaac agtaaatggg ccaacatcac ctggaagcac aatttcaggc
     3721 ctggaactga ctttgtggtt gagtacatcg acagcaacca tacgaaaaaa actgtccctg
     3781 ttaaggccca agcccagccc atacagctga cagacctctt tcccgggatg acgtacacgt
     3841 tgcgggtgta ttcccgggac aacgagggca tcagcagtac cgtcatcacc tttatgacca
     3901 gtacagctta caccaacaac cagactgaca tcgccaccca gggctggttc atcgggctca
     3961 tgtgtgccat tgcccttctg gtgctgatcc tgcttatcgt ctgcttcatc aagaggagtc
     4021 gtggcggcaa gtacccagtg cgagaaaaga aggatgtccc cttgggccct gaagacccca
     4081 aagaagaaga tggttcattt gactacagtg acgaggacaa caagcccctg cagggcagcc
     4141 agacatctct ggacggcacc atcaagcagc aggagagtga cgacagccta gtggactacg
     4201 gcgagggtgg cgagggccag ttcaacgaag atggctcctt tattggccag tacactgtca
     4261 gaaaggacaa ggaggagacc gagggcaatg agagctcaga ggccacatct ccagtcaatg
     4321 ccatctattc cctggcctac ccatacgacg tcccagacta cgcttgagcg gccgcgactc
     4381 tagatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca
     4441 cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt
     4501 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     4561 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta aggcgtaaat
     4621 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     4681 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     4741 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     4801 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
     4861 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     4921 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     4981 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     5041 cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg
     5101 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     5161 ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac
     5221 cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     5281 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     5341 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     5401 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     5461 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     5521 aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg
     5581 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     5641 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     5701 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     5761 tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     5821 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     5881 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     5941 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     6001 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     6061 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     6121 ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     6181 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     6241 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     6301 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     6361 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac
     6421 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
     6481 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     6541 atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata
     6601 ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg
     6661 tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg
     6721 agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt
     6781 tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca
     6841 ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     6901 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     6961 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     7021 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     7081 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     7141 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     7201 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     7261 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     7321 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     7381 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     7441 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     7501 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     7561 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     7621 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     7681 aaccgtatta ccgccatgca ttagttatta atagtaatca attacggggt cattagttca
     7741 tagcccatat atggagttcc gtgttacata acttacggta aatggcccgc ctggctgacc
     7801 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
     7861 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
     7921 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
     7981 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
     8041 cgta
//
