LOCUS       sequence-190343-        8274 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid expressing kinase-dead mutant Nuak1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8274)
  AUTHORS   Lasagna-Reeves CA, de Haro M, Hao S, Park J, Rousseaux MW, Al-Ramahi
            I, Jafar-Nejad P, Vilanova-Velez L, See L, De Maio A, Nitschke L, Wu
            Z, Troncoso JC, Westbrook TF, Tang J, Botas J, Zoghbi HY
  TITLE     Reduction of Nuak1 Decreases Tau and Reverses Phenotypes in a 
            Tauopathy Mouse Model.
  JOURNAL   Neuron. 2016 Oct 19;92(2):407-418. doi: 
            10.1016/j.neuron.2016.09.022. Epub 2016 Oct 6.
  PUBMED    27720485
REFERENCE   2  (bases 1 to 8274)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.neuron.2016.09.022"; journalName: "Neuron"; date: 
            "2016-10-19- 19"; volume: "92"; issue: "2"; pages: "407-418"
COMMENT     Sequence Label: K84M kinase-dead Nuak1-3xflag
FEATURES             Location/Qualifiers
     source          1..8274
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     248..268
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    292..914
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     696..715
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     CDS             complement(2951..3016)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     promoter        complement(3046..3249)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3075..3095)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3250..3629
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     complement(3790..3807)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3790..3806)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3799..3821)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(3947..4402)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4084..4105)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4296..4315
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4429..4533
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4534..4602,4603..5394)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4752..4771)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5565..6153
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6054..6073
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6307..6324
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6441..6462
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6477..6482,6483..6500,6501..6507)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6515..6531
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     6520..6542
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6539..6555
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6539..6555
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6569..6588
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6569..6587
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    6593..6727
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6617..6636)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6671..6690
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(7379..8173)
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(7490..7509)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     8100..8119
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(8227..8274,1..250))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
ORIGIN
        1 gcctcggcct ctgcataaat aaaaaaaatt agtcagccat ggggcggaga atgggcggaa
       61 ctgggcggag ttaggggcgg gatgggcgga gttaggggcg ggactatggt tgctgactaa
      121 ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt tccacacctg
      181 gttgctgact aattgagatg catgctttgc atacttctgc ctgctgggga gcctggggac
      241 tttccacacc ctaactgaca cacattccac agccaagctg gccgcgtacc caattcaaca
      301 ggcatctact gagtggaccc aacgcatgag aggacagtgc caagcaagca actcaaatgt
      361 cccaccggtt gggcatggcc aggtagccta tgctgtgtct ggacgtcctc ctgctggtat
      421 agttatttta aaatcagaag gacagggaag ggagcagtgg ttcacgcctg taatcccagc
      481 aatttgggag gccaaggtgg gtagatcacc tgagattagg agttggagac cagcctggcc
      541 aatatggtga aaccccgtct ctaccaaaaa aacaaaaatt agctgagcct ggtcatgcat
      601 gcctggaatc ccaacaactc gggaggctga ggcaggagaa tcgcttgaac ccaggaggcg
      661 gagattgcag tgagccaaga ttgtgccact gcactccagc ttggttccca atagaccccg
      721 caggccctac aggttgtctt cccaacttgc cccttgctcc ataccacccc cctccacccc
      781 ataatattat agaaggacac ctagtcagac aaaatgatgc aacttaattt tattaggaca
      841 aggctggtgg gcactggagt ggcaacttcc agggccagga gaggcactgg ggaggggtca
      901 cagggatgcc acccgggatc ctctagagtc gactggtacc gatatcagat ctatcgatga
      961 attcctagtt gagcttgctg cagatctcca gcgcttgctt gtagacctga gtcacatcat
     1021 ccatgtctgt gaggagggag aagctgctgt ctgccagccg gttccggtac cgcttcaggt
     1081 actggggccg gggccggttc tggagcccct caaagtcctg gatctggagg aagttttctg
     1141 cagagacgca gctgcggatg cgctggcggg cagggcgatt ctcctgcaaa tccagcaagt
     1201 caaaagagtc gctggacagc acgctgtcat cgctgatgac actggaaggg cggctgtagc
     1261 tccgggagag gccctcggca gggacaccag gctctgacag ggattccagt gtgggcattt
     1321 cagggctgac cagggctggg tccatggtgc ccgctgagta tttgctgctg tgtttcaaga
     1381 tgcccttcct ccggcaggag aggctgtggg aggttaccct ggctgggtcc ggggggctgg
     1441 gggaggggat gctgctgccc atcacatcat tactgtccaa cagctccgaa gactcactgc
     1501 gctctgggga agagtagtaa cctgattctc tctgctgggt ctttttcaag atgcctttct
     1561 tgggcatcgt ggccgactgc ttggggctga gttttcccgg cacctctgcc tctggtgagc
     1621 ttgggaggag cacgccagtc ctgcacaagt cctgctccat cttgaaagta gagggtaagg
     1681 caggaccaac tacaccttca atgaagccag tgctgtgaga gcgatgctcg ctgttgcttc
     1741 gcttcttcag gatccccttg ggcctcttag aactcaactt ggatgggctt tcaggcactg
     1801 catcctgacc agactgagca aagtcattct ctttcttgga tttcttcagc gaccgctgcc
     1861 gctctagcat gacctcagag gtcgtgggtt tggccaggcc cttcattttg gcttcggtgt
     1921 cagcctgcag ccctgtggaa cggtggtgcc agtcaatgat ccgagccagg agtggggact
     1981 cagagtcatg gagggcatca cagtcacaca cgctgctctt atagccccag ttcacccacc
     2041 agtggttggc aatgtcctca atagtggccc ggcgatcggg gttcaccatc agcatccacc
     2101 gtatgagtcc tcgagcatct gagggctgtg ttggctcccg gtactctccg ctgctgattt
     2161 gccgaatgag gtttttgtga tcgaaaccat cgaagggcat tgttccataa acaagagtgt
     2221 aaagcaacac acccagggcc cagctgtcca cctctggccc tcggtaaggt ctcccattga
     2281 caatctcagg agatgcatag agtggactcc cacaaaacgt ttgtaagaac ttatccttct
     2341 ggtacaggtt ggaaagccca aagtcagcaa tcttaatatt gcagttgtca tcgagcagta
     2401 tattttccag cttcaagtcc cggtggacca caccgttctt gtgacaatag tgcacagcag
     2461 agacgatctg ccggaagaag tgccgggtct ccctctcact gaggcgtcgc cgctcactga
     2521 tgtaatcgta cagctcccct ttgctggcat attccatgat gatcacaatc ttatctttgt
     2581 tctcaaacac ttcataaata ctgatgatat gaggatggtt gagagatgac atgatctcaa
     2641 tctctcgtct gatgtgaacc atgtcttgtt catccttaat tttgtcctta cgaatggaca
     2701 ttatcgcaac cactcggcca gaaaacctct cggtggcccg cttgactttg ccgtaggtgc
     2761 ctttgcccag ggtctcctgc agctcgtagc ggtgcttcaa gttgtgcttg tggtgatgcc
     2821 gcttcacccc gtgcggcttc ctgggctcca gggctgcagt cgcccccgcc accgcctctc
     2881 ggggagagcc cggcgccccc agccccaagt cggggcggtc ccccgccaca ggcgcggcgg
     2941 ccccttcaag cttgtcatcg tcatccttgt aatcgatgtc atgatcttta taatcaccgt
     3001 catggtcttt gtagtccatg gttaattctg acggttcact aaacgagctc tgcttatata
     3061 gacctcccac cgtacacgcc taccgcccat ttgcgtcaac ggggcggggt tattacgaca
     3121 ttttggaaag tcccgttgat tttggtgcca aaacaaactc ccattgacgt caatggggtg
     3181 gagacttgga aatccccgtg agtcaaaccg ctatccacgc ccattggtgt actgccaaaa
     3241 ccgcatcacc atggtaatag cgatgactaa tacgtagatg tactgccaag taggaaagtc
     3301 ccgtaaggtc atgtactggg cataatgcca ggcgggccat ttaccgtcat tgacgtcaat
     3361 agggggcgga cttggcatat gatacacttg atgtactgcc aagtgggcag tttaccgtaa
     3421 atactccacc cattgacgtc aatggaaagt ccctattggc gttactatgg gaacatacgt
     3481 cattattgac gtcaatgggc gggggtcgtt gggcggtcag ccaggcgggc catttaccgt
     3541 aagttatgta acgcggaact ccatatatgg gctatgaact aatgaccccg taattgatta
     3601 ctattaataa ctagtcaata atcaatgtca acatggcggt catattggac atgagccaat
     3661 ataaatgtac atattatgat atagatacaa cgtatgcaat ggccaatagc caatattgat
     3721 ttatgctata taaccaatga ctaatatggc taattgccaa tattgattca atgtatagat
     3781 cagcttggca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca
     3841 acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg
     3901 caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggaaat tgtaagcgtt
     3961 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     4021 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     4081 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     4141 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     4201 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     4261 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     4321 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     4381 aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     4441 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     4501 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     4561 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     4621 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     4681 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     4741 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     4801 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     4861 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     4921 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     4981 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     5041 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     5101 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     5161 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     5221 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     5281 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     5341 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     5401 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     5461 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     5521 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     5581 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     5641 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     5701 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     5761 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     5821 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     5881 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     5941 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     6001 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     6061 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     6121 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     6181 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     6241 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     6301 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
     6361 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
     6421 ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta
     6481 cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca
     6541 ggaaacagct atgacatgat tacgaattta atacgactca ctatagggaa ttgatccaga
     6601 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     6661 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     6721 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga
     6781 ggttttttaa agcaagtaaa acctctacaa atgtggtatg gctgattatg atcggcggag
     6841 cctatggaaa aacgccagca acgcggcctt tttacggttc cagcttggcg agtgggtggt
     6901 gagcagctcg ccgtcgcgca gcaggcgcca gcgctccatc cacgcagtga cctcaggcgg
     6961 aaaacgggaa gacacactca tgagatgcct gcaagcaatt cgttctgtat caggcgcagg
     7021 agcgtcccgt ccgggtcgac caaagcggcc atcgtgcctc cccactcctg cagttcgggg
     7081 gcatggatgc gcggatagcc gctgctggtt tcctggatgc cgacggattt gcactgccgg
     7141 tagaactccg cgaggtcgtc cagcctcagg cagcagctga accaactcgc gaggggatcg
     7201 agcccggggt gggcgaagaa ctccagcatg agatccccgc gctggaggat catccagccg
     7261 gcgtcccgga aaacgattcc gaagcccaac ctttcataga aggcggcggt ggaatcgaaa
     7321 tctcgtgatg gcaggttggg cgtcgcttgg tcggtcattt cgaaccccag agtcccgctc
     7381 agaagaactc gtcaagaagg cgatagaagg cgatgcgctg cgaatcggga gcggcgatac
     7441 cgtaaagcac gaggaagcgg tcagcccatt cgccgccaag ctcttcagca atatcacggg
     7501 tagccaacgc tatgtcctga tagcggtccg ccacacccag ccggccacag tcgatgaatc
     7561 cagaaaagcg gccattttcc accatgatat tcggcaagca ggcatcgcca tgggtcacga
     7621 cgagatcctc gccgtcgggc atgcgcgcct tgagcctggc gaacagttcg gctggcgcga
     7681 gcccctgatg ctcttcgtcc agatcatcct gatcgacaag accggcttcc atccgagtac
     7741 gtgctcgctc gatgcgatgt ttcgcttggt ggtcgaatgg gcaggtagcc ggatcaagcg
     7801 tatgcagccg ccgcattgca tcagccatga tggatacttt ctcggcagga gcaaggtgag
     7861 atgacaggag atcctgcccc ggcacttcgc ccaatagcag ccagtccctt cccgcttcag
     7921 tgacaacgtc gagcacagct gcgcaaggaa cgcccgtcgt ggccagccac gatagccgcg
     7981 ctgcctcgtc ctgcagttca ttcagggcac cggacaggtc ggtcttgaca aaaagaaccg
     8041 ggcgcccctg cgctgacagc cggaacacgg cggcatcaga gcagccgatt gtctgttgtg
     8101 cccagtcata gccgaatagc ctctccaccc aagcggccgg agaacctgcg tgcaatccat
     8161 cttgttcaat catgcgaaac gatcctcatc ctgtctcttg atcagatccc aagctgatca
     8221 attcctccaa aaaagcctcc tcactacttc tggaatagct cagaggccga ggcg
//
