LOCUS       sequence-349444-        7642 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mouse DYRK1A tagged with 2A peptide at it's C-terminus and
            mCherry in mammalian cells. Lysine at position 188 substituted with 
            Arginine..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7642)
  AUTHORS   Slepak TI, Salay LD, Lemmon VP, Bixby JL
  TITLE     Dyrk kinases regulate phosphorylation of doublecortin, cytoskeletal 
            organization, and neuronal morphology.
  JOURNAL   Cytoskeleton (Hoboken). 2012 Jul;69(7):514-27. doi: 
            10.1002/cm.21021. Epub 2012 Mar 7.
  PUBMED    22359282
REFERENCE   2  (bases 1 to 7642)
  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: "10.1002/cm.21021"; 
            journalName: "Cytoskeleton (Hoboken)"; date: "2012-07"; volume: 
            "69"; issue: "7"; pages: "514-27"
COMMENT     Sequence Label: pAAV-DYRK1A(K188R)-2A-mCherry
FEATURES             Location/Qualifiers
     source          1..7642
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        214..417
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     368..388
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     414..438
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          566..944
                     /label=chimeric intron
                     /note="fusion between human cytomegalovirus intron A and 
                     human beta-globin intron 2"
     CDS             2813..2842
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             2816..2845
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             2819..2848
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             2822..2851
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             3290..3343
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             3371..4078
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(3513..3531)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3781..3800
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    4093..4569
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4292..4311)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4609..4749
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4609..4738
                     /label=AAV2 ITR
     rep_origin      4824..5279
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4911..4930)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5121..5142
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5296..5315)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5415..5437
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5475..5493)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5561..5665
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5666..5734,5735..6526)
                     /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(5884..5903)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6697..7285
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7186..7205
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   7347..7476
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     enhancer        join(7552..7642,1..213)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
       61 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
      121 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
      181 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
      241 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
      301 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
      361 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctcgt
      421 ttagtgaacc gtcagatcgc ctggagacgc catccacgct gttttgacct ccatagaaga
      481 caccgggacc gatccagcct ccgcggattc gaatcccggc cgggaacggt gcattggaac
      541 gcggattccc cgtgccaaga gtgacgtaag taccgcctat agagtctata ggcccacaaa
      601 aaatgctttc ttcttttaat atactttttt gtttatctta tttctaatac tttccctaat
      661 ctctttcttt cagggcaata atgatacaat gtatcatgcc tctttgcacc attctaaaga
      721 ataacagtga taatttctgg gttaaggcaa tagcaatatt tctgcatata aatatttctg
      781 catataaatt gtaactgatg taagaggttt catattgcta atagcagcta caatccagct
      841 accattctgc ttttatttta tggttgggat aaggctggat tattctgagt ccaagctagg
      901 cccttttgct aatcatgttc atacctctta tcttcctccc acagctcctg ggcaacgtgc
      961 tggtctgtgt gctggcccat cactttggca aagaattggg attcgaacat cgattgaatt
     1021 catgcataca ggaggagaga cttcagcatg caaaccttca tctgtccggc ttgcaccgtc
     1081 gttctcattc catgctgctg gccttcagat ggctgcacag atgccccact cacaccagta
     1141 cagtgaccgt cgccagccga gcataagtga ccagcaggtg tctgccttac catattctga
     1201 ccagattcag caacctctaa ctaaccagag gcggatgccc caaaccttcc gtgatccagc
     1261 aactgctcct ctgagaaaac tctctgtgga cttgatcaaa acatacaagc atattaatga
     1321 ggtttactat gcaaaaaaga agcgaagaca ccaacagggc cagggggacg attccagtca
     1381 taagaaggag cggaaggttt acaatgatgg ttacgatgat gataactatg attatattgt
     1441 aaaaaacggg gaaaagtgga tggatcggta tgaaatcgac tccttaatag gcaaaggttc
     1501 atttggacag gttgtgaaag cttatgacag agtggagcaa gaatgggtcg ccattagaat
     1561 catcaagaac aagaaagcgt ttctgaatca agcccagata gaagtgcggc tgcttgagct
     1621 catgaacaaa cacgacactg aaatgaagta ctacatagtg catttgaaac gccactttat
     1681 gtttcgaaac catctctgtt tagtgtttga aatgctgtcc tataatctct atgatttgtt
     1741 gagaaacacc aacttccgag gggtctcttt gaacctaaca cgaaagtttg cgcaacagat
     1801 gtgcacagca ttgctttttc ttgcgactcc agaacttagt atcattcact gtgacttaaa
     1861 gcctgagaac atccttctgt gtaaccccaa acggagtgca atcaagattg ttgattttgg
     1921 cagctcttgt cagttggggc agaggatata ccagtatatt cagagtcgct tttatcggtc
     1981 tccagaggtg ctactgggaa tgccttatga ccttgctatc gacatgtggt cccttggatg
     2041 tatcttggtt gaaatgcaca ctggagagcc tctgttcagt ggtgccaatg aggtcgatca
     2101 gatgaataaa atagtggaag tcttgggcat cccacctgct catattcttg accaagcacc
     2161 gaaagcaaga aagttctttg agaagttgcc cgatggcact tggagcttaa agaagaccaa
     2221 agatggaaaa cgggagtaca aaccaccagg aacccgtaaa cttcataata ttcttggagt
     2281 agaaacagga ggacctggcg ggcggcgtgc tggggaatcg ggtcatactg tagctgacta
     2341 cttgaagttc aaagacctca ttttaaggat gcttgattat gaccccaaaa ctcggattca
     2401 accttattat gccctgcagc acagtttttt caagaaaaca gctgatgaag gtaccaacac
     2461 aagtaacagt gtgtctacca gccctgcgat ggagcagtct cagtcttcag gcaccacctc
     2521 cagcacctcc tccagctcag gtggatcctc gggaacgagt aacagtggga gagccaggtc
     2581 ggatccgacg caccagcatc gacacagcgg tggacacttc gctgctgctg tccaggccat
     2641 ggactgtgag acacacagtc cccaggtgcg ccagcagttt ccggctcctc tgggatggtc
     2701 aggcactgaa gctcctacac aagtcactgt tgaaactcat cctgttcaag agacaacctt
     2761 tcatgtagcc ccccagcaga acgcattgca tcatcaccat ggaaacagtt cccatcacca
     2821 ccaccaccac catcaccacc accaccacca tggacagcaa gccttgggta accggaccag
     2881 gccaagggtc tacaattctc caacaaatag ctcctctacc caggattcta tggaggttgg
     2941 ccacagtcac cactccatga catccctgtc ttcctcaaca acttcttcct cgacatcttc
     3001 ctcctctact ggtaatcaag gcaatcaggc ctatcagaac cgcccagtgg ctgctaacac
     3061 cttggacttt ggacagaatg gagctatgga cgttaatttg accgtctact ccaatccccg
     3121 ccaagagact ggcatagctg gacatccaac ataccaattt tctgctaata caggtcctgc
     3181 acattacatg actgaaggac atctggcgat gagacaaggg gctgatagag aagagtctcc
     3241 catgacagga gtttgtgtgc aacagagtcc tgtagctagc tcgaccggtg aaggccgcgg
     3301 cagcctgctg acctgcggcg atgtggaaga aaacccgggc ccggcgccgg gcagcactag
     3361 tcggaagctt gtgagcaagg gcgaggagga taacatggcc atcatcaagg agttcatgcg
     3421 cttcaaggtg cacatggagg gctccgtgaa cggccacgag ttcgagatcg agggcgaggg
     3481 cgagggccgc ccctacgagg gcacccagac cgccaagctg aaggtgacca agggtggccc
     3541 cctgcccttc gcctgggaca tcctgtcccc tcagttcatg tacggctcca aggcctacgt
     3601 gaagcacccc gccgacatcc ccgactactt gaagctgtcc ttccccgagg gcttcaagtg
     3661 ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc gtgacccagg actcctccct
     3721 gcaggacggc gagttcatct acaaggtgaa gctgcgcggc accaacttcc cctccgacgg
     3781 ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc tccgagcgga tgtaccccga
     3841 ggacggcgcc ctgaagggcg agatcaagca gaggctgaag ctgaaggacg gcggccacta
     3901 cgacgctgag gtcaagacca cctacaaggc caagaagccc gtgcagctgc ccggcgccta
     3961 caacgtcaac atcaagttgg acatcacctc ccacaacgag gactacacca tcgtggaaca
     4021 gtacgaacgc gccgagggcc gccactccac cggcggcatg gacgagctgt acaagtaact
     4081 cgagagatct acgggtggca tccctgtgac ccctccccag tgcctctcct ggccctggaa
     4141 gttgccactc cagtgcccac cagccttgtc ctaataaaat taagttgcat cattttgtct
     4201 gactaggtgt ccttctataa tattatgggg tggagggggg tggtatggag caaggggcaa
     4261 gttgggaaga caacctgtag ggcctgcggg gtctattggg aaccaagctg gagtgcagtg
     4321 gcacaatctt ggctcactgc aatctccgcc tcctgggttc aagcgattct cctgcctcag
     4381 cctcccgagt tgttgggatt ccaggcatgc atgaccaggc tcagctaatt tttgtttttt
     4441 tggtagagac ggggtttcac catattggcc aggctggtct ccaactccta atctcaggtg
     4501 atctacccac cttggcctcc caaattgctg ggattacagg cgtgaaccac tgctcccttc
     4561 cctgtccttc tgattttgta ggtaaccacg tgcggaccga gcggccgcag gaacccctag
     4621 tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa
     4681 aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct
     4741 gcctgcaggg gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     4801 gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     4861 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc
     4921 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     4981 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt
     5041 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     5101 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaactctat
     5161 ctcgggctat tcttttgatt tataagggat tttgccgatt tcggtctatt ggttaaaaaa
     5221 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt
     5281 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc
     5341 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     5401 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     5461 cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     5521 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     5581 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     5641 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     5701 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     5761 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     5821 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     5881 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     5941 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     6001 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     6061 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     6121 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     6181 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     6241 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     6301 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     6361 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     6421 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     6481 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     6541 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     6601 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     6661 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     6721 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     6781 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     6841 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     6901 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     6961 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     7021 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     7081 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     7141 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     7201 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     7261 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     7321 cttttgctgg ccttttgctc acatgtcctg caggcagctg cgcgctcgct cgctcactga
     7381 ggccgcccgg gcgtcgggcg acctttggtc gcccggcctc agtgagcgag cgagcgcgca
     7441 gagagggagt ggccaactcc atcactaggg gttcctgcgg ccgcacgcgt ctagttatta
     7501 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
     7561 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
     7621 aatgacgtat gttcccatag ta
//
