LOCUS       sequence-477732-        8044 bp    DNA     circular SYN 03-JUL-2025
DEFINITION  Expresses rat NF186 with a TAG codon at position 519 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(K519TAG)-HA
FEATURES             Location/Qualifiers
     source          1..8044
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        62..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             4681..4707
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    4832..4953
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4869..4888)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4923..4942
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4960..5415)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5097..5118)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5309..5328
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5442..5546
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5548..5905
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5548..5739
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5571..5591)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5756..5891
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5940..6734
                     /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             5940..6734
                     /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(5994..6013)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6604..6623
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6922..6941)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6966..7013
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7342..7930
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7831..7850
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 tgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagct caagcttgat atcgaattcc gggcggcggc tggcggcgag
      661 agctgctgca ggggacgcgg gcgaagtggc ggcgccggga gcagacatct gggactgcgc
      721 ccagggctgg agctggagct cctggaggta aggttgactg acttatgtgc gatttgggag
      781 cctggagttt tcttgccctc tgccgctgcc gtcaaccgga gcagagcctc cgctggtgtt
      841 gcgaaaagca ggccgaatga ggcagagaag ctgagcactg tctggaaagc ccacgaggaa
      901 cggcgcaagg catcgagagc ccgtgggctg ggcagtgcgg agcagggcct ggcgctgagg
      961 atggccaggc agcaggcgcc accctgggtc cacgtagccc tcatcctctt cctcctcagc
     1021 ctcggagggg ccattgagat tccgatggat ccaagcattc agaatgagct gacccaaccc
     1081 ccaacgatca ccaagcagtc ggtgaaggac cacatcgtgg acccccgaga taacatcctg
     1141 attgaatgtg aagctaaagg gaaccccgcc cccagtttcc actggactcg caacagcagg
     1201 ttcttcaaca ttgccaagga cccacgggtg tccatgagga ggaggtctgg gaccttggtg
     1261 atcgacttcc gcagtggtgg gcggcctgag gagtacgaag gggagtacca gtgctttgcc
     1321 cggaacaaat tcggcacagc tcttagcaac cgcatccgcc tgcaggtgtc caaatctccc
     1381 ctgtggccca aggaaaacct agaccccgtc gtggttcaag agggtgcccc cttaaccctg
     1441 cagtgcaacc ccccacctgg cctcccatcc cccgtcatct tctggatgag cagctccatg
     1501 gagcccatca cccaggacaa gcgtgtctcc cagggtcaca acggggacct gtacttctcc
     1561 aacgtcatgc tgcaggacat gcagaccgac tacagctgca atgcacgctt ccacttcacc
     1621 cacaccattc agcagaagaa tcccttcacc ctcaaagtcc tcaccacccg aggagttgcg
     1681 gaaagaacac ctagcttcat gtatccccag ggcacgtcaa gcagtcagat ggtactgcgc
     1741 ggcatggacc tgctgctgga gtgcattgcc tctggggtcc caacaccaga tattgcatgg
     1801 tacaagaaag gtggggacct cccatctgac aaggccaagt tcgagaattt taacaaggct
     1861 ctgcgcatca ccaatgtctc tgaagaggac tctggggagt atttctgcct ggcctccaac
     1921 aagatgggca gcatccggca cacgatctcg gtgagagtaa aggctgcccc atactggctg
     1981 gatgagccca agaacttgat cctggctcct ggtgaagatg ggaggctggt gtgtcgagcc
     2041 aatgggaacc cgaagccgac cgtccagtgg ctggtgaatg gagacccttt gcaatcggca
     2101 ccacccaacc ccaaccgtga ggtggccgga gacactatca tcttccggga cactcagatc
     2161 agcagcaggg cagtgtacca gtgtaacaca tccaacgaac atggctacct gctggccaat
     2221 gccttcgtca gcgtattaga tgtaccccct cggatgctgt ctccccggaa ccagctcatc
     2281 agggtgatcc tttacaaccg gacgcgactg gactgtccgt tctttgggtc tcccattcca
     2341 acactccgat ggtttaagaa tgggcaagga agcaacctgg atggtggtaa ctaccacgtc
     2401 tacgaaaatg gcagcctgga aatcaagatg attcgcaaag aggaccaagg catctacacc
     2461 tgtgtggcca ccaacatcct gggaaaagct gaaaatcaag tccgcctgga ggtctaggac
     2521 cccaccagga tctacaggat gcctgaagac caggtggcca agaggggcac cacagtgcag
     2581 ctggagtgcc gtgtgaagca tgacccctcc ttgaaactca cagtctcctg gctgaaggat
     2641 gacgagccac tctacattgg aaacaggatg aaaaaggaag atgactccct gaccatcttc
     2701 ggagtggcag agcgggacca aggcagttac acgtgcatgg ccagcaccga gctggaccag
     2761 gacctggcaa aggcctacct cactgttcta gctgatcagg ccactccgac taaccgtttg
     2821 gctgccctgc ccaaagggcg gccagaccga cccagggacc tggagctcac tgacctggcg
     2881 gaaaggagtg tgaggctgac ctggatcccg ggggatgata acaacagtcc catcacagac
     2941 tacgttgtcc agtttgaaga ggaccagttc cagccaggag tctggcatga ccactccaag
     3001 ttcccaggca gtgtcaactc agccgtcctc catctgtccc cgtatgtcaa ctatcagttc
     3061 cgagtcatcg ctgtcaacga ggttgggagc agccacccca gccttccatc cgagcggtac
     3121 cgaacaagcg gggcaccccc tgaatccaac cccagtgatg tgaagggcga agggacacga
     3181 aaaaacaata tggagatcac atggacgcct atgaatgcta cctccgcctt tggccccaac
     3241 ctgcgctaca ttgtcaagtg gcgacggaga gaaacccgag agacttggaa caatgtcacc
     3301 gtgtggggct ctcgctatgt ggtggggcag acccctgtct acgtacccta tgagatccga
     3361 gtccaggctg aaaatgactt tgggaaaggc ccggagcctg aaaccgtcat tgggtactcg
     3421 ggggaagatt tacccagtgc ccccaggcgt ttcagagtcc gacagcccaa cctggagacc
     3481 atcaacctgg aatgggatca cccagagcac cccaacggga tcctgattgg atacacgctc
     3541 agatacgtgc cctttaatgg aaccaaactg ggaaagcaga tggtggaaaa cttctctccc
     3601 aatcagacca agttctcggt gcagagagca gaccccgtgt cccgttaccg cttctccctc
     3661 agtgccagga cgcaggtggg ctctggagaa gcagccacgg aggagtcccc aacacctcca
     3721 aatgaagcta ctccaactgc agctcctccc acattgcccc cgactaccgt gggtaccaca
     3781 ggccttgtga gcagtactga tgctactgcc cttgctgcca ccagtgaagc cacaacagtt
     3841 cccatcatcc caaccgtcgt acctaccacc gtcgccacca ccattgccac aactactaca
     3901 accactgccg ccgccaccac cactaccacc acggagagtc ctcccactac caccactggg
     3961 actaagattc acgaaaccgc ccccgacgag cagtccattt ggaacgtcac ggtgctcccc
     4021 aacagtaaat gggccaacat cacctggaag cacaatttca ggcctggaac tgactttgtg
     4081 gttgagtaca tcgacagcaa ccatacgaaa aaaactgtcc ctgttaaggc ccaagcccag
     4141 cccatacagc tgacagacct ctttcccggg atgacgtaca cgttgcgggt gtattcccgg
     4201 gacaacgagg gcatcagcag taccgtcatc acctttatga ccagtacagc ttacaccaac
     4261 aaccagactg acatcgccac ccagggctgg ttcatcgggc tcatgtgtgc cattgccctt
     4321 ctggtgctga tcctgcttat cgtctgcttc atcaagagga gtcgtggcgg caagtaccca
     4381 gtgcgagaaa agaaggatgt ccccttgggc cctgaagacc ccaaagaaga agatggttca
     4441 tttgactaca gtgacgagga caacaagccc ctgcagggca gccagacatc tctggacggc
     4501 accatcaagc agcaggagag tgacgacagc ctagtggact acggcgaggg tggcgagggc
     4561 cagttcaacg aagatggctc ctttattggc cagtacactg tcagaaagga caaggaggag
     4621 accgagggca atgagagctc agaggccaca tctccagtca atgccatcta ttccctggcc
     4681 tacccatacg acgtcccaga ctacgcttga gcggccgcga ctctagatca taatcagcca
     4741 taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct
     4801 gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta
     4861 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     4921 ttgtggtttg tccaaactca tcaatgtatc ttaaggcgta aattgtaagc gttaatattt
     4981 tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa
     5041 tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag
     5101 tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg
     5161 tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga
     5221 ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg
     5281 gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg
     5341 cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc
     5401 cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     5461 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     5521 ttcaataata ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg
     5581 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     5641 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     5701 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     5761 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     5821 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     5881 ttttggaggc ctaggctttt gcaaagatcg atcaagagac aggatgagga tcgtttcgca
     5941 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     6001 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     6061 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     6121 aagacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     6181 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     6241 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     6301 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     6361 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     6421 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgagc atgcccgacg
     6481 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     6541 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     6601 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     6661 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     6721 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct
     6781 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     6841 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     6901 ccaccctagg gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc
     6961 tatgacggca ataaaaagac agaataaaac gcacggtgtt gggtcgtttg ttcataaacg
     7021 cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca
     7081 atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg
     7141 gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc tcaggttact catatatact
     7201 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     7261 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     7321 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     7381 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     7441 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     7501 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     7561 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     7621 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     7681 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     7741 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     7801 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     7861 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     7921 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     7981 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat
     8041 gcat
//
