LOCUS       sequence-223418-        7665 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mouse DYRK1A tagged with 2A peptide at it's C-terminus and
            GFP in mammalian cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7665)
  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 7665)
  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-2A-GFP
FEATURES             Location/Qualifiers
     source          1..7665
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     enhancer        221..524
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        525..727
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     678..698
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     724..748
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          876..1254
                     /label=chimeric intron
                     /note="fusion between human cytomegalovirus intron A and 
                     human beta-globin intron 2"
     CDS             3123..3152
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             3126..3155
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             3129..3158
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             3132..3161
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             3600..3653
                     /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             join(3681..3683,3684..4397)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(3681..3683,3684..4394)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(3723..3744)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3984..4003)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4331..4352
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4412..4888
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4611..4630)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4928..5068
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4928..5057
                     /label=AAV2 ITR
     rep_origin      5143..5598
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5230..5249)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5440..5461
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5615..5634)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5734..5756
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5794..5812)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5880..5984
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5985..6053,6054..6845)
                     /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(6203..6222)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7016..7604
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7505..7524
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtggagc tagttattaa tagtaatcaa
      181 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
      241 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
      301 ttcccatagt aacgtcaata gggactttcc attgacgtca atgggtggag tatttacggt
      361 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
      421 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
      481 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
      541 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
      601 ttgacgtcaa tgggagtttg ttttgcacca aaatcaacgg gactttccaa aatgtcgtaa
      661 caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
      721 cagagctcgt ttagtgaacc gtcagatcgc ctggagacgc catccacgct gttttgacct
      781 ccatagaaga caccgggacc gatccagcct ccgcggattc gaatcccggc cgggaacggt
      841 gcattggaac gcggattccc cgtgccaaga gtgacgtaag taccgcctat agagtctata
      901 ggcccacaaa aaatgctttc ttcttttaat atactttttt gtttatctta tttctaatac
      961 tttccctaat ctctttcttt cagggcaata atgatacaat gtatcatgcc tctttgcacc
     1021 attctaaaga ataacagtga taatttctgg gttaaggcaa tagcaatatt tctgcatata
     1081 aatatttctg catataaatt gtaactgatg taagaggttt catattgcta atagcagcta
     1141 caatccagct accattctgc ttttatttta tggttgggat aaggctggat tattctgagt
     1201 ccaagctagg cccttttgct aatcatgttc atacctctta tcttcctccc acagctcctg
     1261 ggcaacgtgc tggtctgtgt gctggcccat cactttggca aagaattggg attcgaacat
     1321 cgattgaatt catgcataca ggaggagaga cttcagcatg caaaccttca tctgtccggc
     1381 ttgcaccgtc gttctcattc catgctgctg gccttcagat ggctgcacag atgccccact
     1441 cacaccagta cagtgaccgt cgccagccga gcataagtga ccagcaggtg tctgccttac
     1501 catattctga ccagattcag caacctctaa ctaaccagag gcggatgccc caaaccttcc
     1561 gtgatccagc aactgctcct ctgagaaaac tctctgtgga cttgatcaaa acatacaagc
     1621 atattaatga ggtttactat gcaaaaaaga agcgaagaca ccaacagggc cagggggacg
     1681 attccagtca taagaaggag cggaaggttt acaatgatgg ttacgatgat gataactatg
     1741 attatattgt aaaaaacggg gaaaagtgga tggatcggta tgaaatcgac tccttaatag
     1801 gcaaaggttc atttggacag gttgtgaaag cttatgacag agtggagcaa gaatgggtcg
     1861 ccattaaaat catcaagaac aagaaagcgt ttctgaatca agcccagata gaagtgcggc
     1921 tgcttgagct catgaacaaa cacgacactg aaatgaagta ctacatagtg catttgaaac
     1981 gccactttat gtttcgaaac catctctgtt tagtgtttga aatgctgtcc tataatctct
     2041 atgatttgtt gagaaacacc aacttccgag gggtctcttt gaacctaaca cgaaagtttg
     2101 cgcaacagat gtgcacagca ttgctttttc ttgcgactcc agaacttagt atcattcact
     2161 gtgacttaaa gcctgagaac atccttctgt gtaaccccaa acggagtgca atcaagattg
     2221 ttgattttgg cagctcttgt cagttggggc agaggatata ccagtatatt cagagtcgct
     2281 tttatcggtc tccagaggtg ctactgggaa tgccttatga ccttgctatc gacatgtggt
     2341 cccttggatg tatcttggtt gaaatgcaca ctggagagcc tctgttcagt ggtgccaatg
     2401 aggtcgatca gatgaataaa atagtggaag tcttgggcat cccacctgct catattcttg
     2461 accaagcacc gaaagcaaga aagttctttg agaagttgcc cgatggcact tggagcttaa
     2521 agaagaccaa agatggaaaa cgggagtaca aaccaccagg aacccgtaaa cttcataata
     2581 ttcttggagt agaaacagga ggacctggcg ggcggcgtgc tggggaatcg ggtcatactg
     2641 tagctgacta cttgaagttc aaagacctca ttttaaggat gcttgattat gaccccaaaa
     2701 ctcggattca accttattat gccctgcagc acagtttttt caagaaaaca gctgatgaag
     2761 gtaccaacac aagtaacagt gtgtctacca gccctgcgat ggagcagtct cagtcttcag
     2821 gcaccacctc cagcacctcc tccagctcag gtggatcctc gggaacgagt aacagtggga
     2881 gagccaggtc ggatccgacg caccagcatc gacacagcgg tggacacttc gctgctgctg
     2941 tccaggccat ggactgtgag acacacagtc cccaggtgcg ccagcagttt ccggctcctc
     3001 tgggatggtc aggcactgaa gctcctacac aagtcactgt tgaaactcat cctgttcaag
     3061 agacaacctt tcatgtagcc ccccagcaga acgcattgca tcatcaccat ggaaacagtt
     3121 cccatcacca ccaccaccac catcaccacc accaccacca tggacagcaa gccttgggta
     3181 accggaccag gccaagggtc tacaattctc caacaaatag ctcctctacc caggattcta
     3241 tggaggttgg ccacagtcac cactccatga catccctgtc ttcctcaaca acttcttcct
     3301 cgacatcttc ctcctctact ggtaatcaag gcaatcaggc ctatcagaac cgcccagtgg
     3361 ctgctaacac cttggacttt ggacagaatg gagctatgga cgttaatttg accgtctact
     3421 ccaatccccg ccaagagact ggcatagctg gacatccaac ataccaattt tctgctaata
     3481 caggtcctgc acattacatg actgaaggac atctggcgat gagacaaggg gctgatagag
     3541 aagagtctcc catgacagga gtttgtgtgc aacagagtcc tgtagctagc tcgaccggtg
     3601 aaggccgcgg cagcctgctg acctgcggcg atgtggaaga aaacccgggc ccggcgccgg
     3661 gcagcactag tcggaagctt gtgagcaagg gcgaggagct gttcaccggg gtggtgccca
     3721 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg
     3781 agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc
     3841 ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct
     3901 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc
     3961 aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt
     4021 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg
     4081 gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg
     4141 ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg
     4201 gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc
     4261 tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga
     4321 agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg
     4381 acgagctgta caagtaactc gagagatcta cgggtggcat ccctgtgacc cctccccagt
     4441 gcctctcctg gccctggaag ttgccactcc agtgcccacc agccttgtcc taataaaatt
     4501 aagttgcatc attttgtctg actaggtgtc cttctataat attatggggt ggaggggggt
     4561 ggtatggagc aaggggcaag ttgggaagac aacctgtagg gcctgcgggg tctattggga
     4621 accaagctgg agtgcagtgg cacaatcttg gctcactgca atctccgcct cctgggttca
     4681 agcgattctc ctgcctcagc ctcccgagtt gttgggattc caggcatgca tgaccaggct
     4741 cagctaattt ttgttttttt ggtagagacg gggtttcacc atattggcca ggctggtctc
     4801 caactcctaa tctcaggtga tctacccacc ttggcctccc aaattgctgg gattacaggc
     4861 gtgaaccact gctcccttcc ctgtccttct gattttgtag gtaaccacgt gcggaccgag
     4921 cggccgcagg aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc
     4981 actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg
     5041 agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc ggtattttct ccttacgcat
     5101 ctgtgcggta tttcacaccg catacgtcaa agcaaccata gtacgcgccc tgtagcggcg
     5161 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     5221 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     5281 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     5341 accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     5401 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     5461 gaacaacact caaccctatc tcgggctatt cttttgattt ataagggatt ttgccgattt
     5521 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     5581 tattaacgtt tacaatttta tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     5641 taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
     5701 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
     5761 caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg
     5821 ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc
     5881 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     5941 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     6001 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     6061 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     6121 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     6181 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     6241 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     6301 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     6361 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     6421 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     6481 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     6541 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     6601 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     6661 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     6721 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     6781 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     6841 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     6901 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     6961 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     7021 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     7081 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     7141 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
     7201 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     7261 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     7321 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     7381 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     7441 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     7501 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     7561 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     7621 cctttttacg gttcctggcc ttttgctggc cttttgctca catgt
//
