LOCUS       sequence-477520-        8044 bp    DNA     circular SYN 01-JUL-2025
DEFINITION  Expresses rat NF186 with a TAG codon at position 604 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(K604TAG)-HA
FEATURES             Location/Qualifiers
     source          1..8044
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        82..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        385..588
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     539..559
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             4701..4727
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    4852..4973
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4889..4908)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4943..4962
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4980..5435)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5117..5138)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5329..5348
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5462..5566
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5568..5925
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5568..5759
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5591..5611)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5776..5911
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5960..6754
                     /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             5960..6754
                     /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(6014..6033)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6624..6643
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6942..6961)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6986..7033
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7362..7950
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7851..7870
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 accgtattac cgccatgcat tagttattaa tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg tgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga
      421 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
      481 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
      541 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctgg tttagtgaac
      601 cgtcagatcc gctagcgcta ccggactcag atctcgagct caagcttgat atcgaattcc
      661 gggcggcggc tggcggcgag agctgctgca ggggacgcgg gcgaagtggc ggcgccggga
      721 gcagacatct gggactgcgc ccagggctgg agctggagct cctggaggta aggttgactg
      781 acttatgtgc gatttgggag cctggagttt tcttgccctc tgccgctgcc gtcaaccgga
      841 gcagagcctc cgctggtgtt gcgaaaagca ggccgaatga ggcagagaag ctgagcactg
      901 tctggaaagc ccacgaggaa cggcgcaagg catcgagagc ccgtgggctg ggcagtgcgg
      961 agcagggcct ggcgctgagg atggccaggc agcaggcgcc accctgggtc cacgtagccc
     1021 tcatcctctt cctcctcagc ctcggagggg ccattgagat tccgatggat ccaagcattc
     1081 agaatgagct gacccaaccc ccaacgatca ccaagcagtc ggtgaaggac cacatcgtgg
     1141 acccccgaga taacatcctg attgaatgtg aagctaaagg gaaccccgcc cccagtttcc
     1201 actggactcg caacagcagg ttcttcaaca ttgccaagga cccacgggtg tccatgagga
     1261 ggaggtctgg gaccttggtg atcgacttcc gcagtggtgg gcggcctgag gagtacgaag
     1321 gggagtacca gtgctttgcc cggaacaaat tcggcacagc tcttagcaac cgcatccgcc
     1381 tgcaggtgtc caaatctccc ctgtggccca aggaaaacct agaccccgtc gtggttcaag
     1441 agggtgcccc cttaaccctg cagtgcaacc ccccacctgg cctcccatcc cccgtcatct
     1501 tctggatgag cagctccatg gagcccatca cccaggacaa gcgtgtctcc cagggtcaca
     1561 acggggacct gtacttctcc aacgtcatgc tgcaggacat gcagaccgac tacagctgca
     1621 atgcacgctt ccacttcacc cacaccattc agcagaagaa tcccttcacc ctcaaagtcc
     1681 tcaccacccg aggagttgcg gaaagaacac ctagcttcat gtatccccag ggcacgtcaa
     1741 gcagtcagat ggtactgcgc ggcatggacc tgctgctgga gtgcattgcc tctggggtcc
     1801 caacaccaga tattgcatgg tacaagaaag gtggggacct cccatctgac aaggccaagt
     1861 tcgagaattt taacaaggct ctgcgcatca ccaatgtctc tgaagaggac tctggggagt
     1921 atttctgcct ggcctccaac aagatgggca gcatccggca cacgatctcg gtgagagtaa
     1981 aggctgcccc atactggctg gatgagccca agaacttgat cctggctcct ggtgaagatg
     2041 ggaggctggt gtgtcgagcc aatgggaacc cgaagccgac cgtccagtgg ctggtgaatg
     2101 gagacccttt gcaatcggca ccacccaacc ccaaccgtga ggtggccgga gacactatca
     2161 tcttccggga cactcagatc agcagcaggg cagtgtacca gtgtaacaca tccaacgaac
     2221 atggctacct gctggccaat gccttcgtca gcgtattaga tgtaccccct cggatgctgt
     2281 ctccccggaa ccagctcatc agggtgatcc tttacaaccg gacgcgactg gactgtccgt
     2341 tctttgggtc tcccattcca acactccgat ggtttaagaa tgggcaagga agcaacctgg
     2401 atggtggtaa ctaccacgtc tacgaaaatg gcagcctgga aatcaagatg attcgcaaag
     2461 aggaccaagg catctacacc tgtgtggcca ccaacatcct gggaaaagct gaaaatcaag
     2521 tccgcctgga ggtcaaagac cccaccagga tctacaggat gcctgaagac caggtggcca
     2581 agaggggcac cacagtgcag ctggagtgcc gtgtgaagca tgacccctcc ttgaaactca
     2641 cagtctcctg gctgaaggat gacgagccac tctacattgg aaacaggatg aaaaaggaag
     2701 atgactccct gaccatcttc ggagtggcag agcgggacca aggcagttac acgtgcatgg
     2761 ccagcaccga gctggaccag gacctggcat aggcctacct cactgttcta gctgatcagg
     2821 ccactccgac taaccgtttg gctgccctgc ccaaagggcg gccagaccga cccagggacc
     2881 tggagctcac tgacctggcg gaaaggagtg tgaggctgac ctggatcccg ggggatgata
     2941 acaacagtcc catcacagac tacgttgtcc agtttgaaga ggaccagttc cagccaggag
     3001 tctggcatga ccactccaag ttcccaggca gtgtcaactc agccgtcctc catctgtccc
     3061 cgtatgtcaa ctatcagttc cgagtcatcg ctgtcaacga ggttgggagc agccacccca
     3121 gccttccatc cgagcggtac cgaacaagcg gggcaccccc tgaatccaac cccagtgatg
     3181 tgaagggcga agggacacga aaaaacaata tggagatcac atggacgcct atgaatgcta
     3241 cctccgcctt tggccccaac ctgcgctaca ttgtcaagtg gcgacggaga gaaacccgag
     3301 agacttggaa caatgtcacc gtgtggggct ctcgctatgt ggtggggcag acccctgtct
     3361 acgtacccta tgagatccga gtccaggctg aaaatgactt tgggaaaggc ccggagcctg
     3421 aaaccgtcat tgggtactcg ggggaagatt tacccagtgc ccccaggcgt ttcagagtcc
     3481 gacagcccaa cctggagacc atcaacctgg aatgggatca cccagagcac cccaacggga
     3541 tcctgattgg atacacgctc agatacgtgc cctttaatgg aaccaaactg ggaaagcaga
     3601 tggtggaaaa cttctctccc aatcagacca agttctcggt gcagagagca gaccccgtgt
     3661 cccgttaccg cttctccctc agtgccagga cgcaggtggg ctctggagaa gcagccacgg
     3721 aggagtcccc aacacctcca aatgaagcta ctccaactgc agctcctccc acattgcccc
     3781 cgactaccgt gggtaccaca ggccttgtga gcagtactga tgctactgcc cttgctgcca
     3841 ccagtgaagc cacaacagtt cccatcatcc caaccgtcgt acctaccacc gtcgccacca
     3901 ccattgccac aactactaca accactgccg ccgccaccac cactaccacc acggagagtc
     3961 ctcccactac caccactggg actaagattc acgaaaccgc ccccgacgag cagtccattt
     4021 ggaacgtcac ggtgctcccc aacagtaaat gggccaacat cacctggaag cacaatttca
     4081 ggcctggaac tgactttgtg gttgagtaca tcgacagcaa ccatacgaaa aaaactgtcc
     4141 ctgttaaggc ccaagcccag cccatacagc tgacagacct ctttcccggg atgacgtaca
     4201 cgttgcgggt gtattcccgg gacaacgagg gcatcagcag taccgtcatc acctttatga
     4261 ccagtacagc ttacaccaac aaccagactg acatcgccac ccagggctgg ttcatcgggc
     4321 tcatgtgtgc cattgccctt ctggtgctga tcctgcttat cgtctgcttc atcaagagga
     4381 gtcgtggcgg caagtaccca gtgcgagaaa agaaggatgt ccccttgggc cctgaagacc
     4441 ccaaagaaga agatggttca tttgactaca gtgacgagga caacaagccc ctgcagggca
     4501 gccagacatc tctggacggc accatcaagc agcaggagag tgacgacagc ctagtggact
     4561 acggcgaggg tggcgagggc cagttcaacg aagatggctc ctttattggc cagtacactg
     4621 tcagaaagga caaggaggag accgagggca atgagagctc agaggccaca tctccagtca
     4681 atgccatcta ttccctggcc tacccatacg acgtcccaga ctacgcttga gcggccgcga
     4741 ctctagatca taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc
     4801 ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt
     4861 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     4921 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaaggcgta
     4981 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     5041 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     5101 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     5161 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
     5221 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     5281 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     5341 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     5401 acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt
     5461 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     5521 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtcctg aggcggaaag
     5581 aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     5641 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     5701 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     5761 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     5821 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     5881 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac
     5941 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc
     6001 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     6061 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     6121 cggtgccctg aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg
     6181 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     6241 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     6301 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     6361 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     6421 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     6481 caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     6541 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     6601 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     6661 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     6721 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc
     6781 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
     6841 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     6901 ctcatgctgg agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca
     6961 ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt
     7021 gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca
     7081 ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca
     7141 agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc
     7201 tcaggttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     7261 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     7321 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     7381 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     7441 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     7501 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     7561 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     7621 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     7681 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     7741 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     7801 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     7861 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     7921 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     7981 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     8041 gata
//
