LOCUS       sequence-229494-        7651 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 7651)
  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 7651)
  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..7651
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     enhancer        206..509
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        510..713
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     664..684
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     710..734
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          862..1240
                     /label=chimeric intron
                     /note="fusion between human cytomegalovirus intron A and 
                     human beta-globin intron 2"
     CDS             3109..3138
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             3112..3141
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             3115..3144
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             3118..3147
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             3586..3639
                     /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(3667..3669,3670..4383)
                     /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(3667..3669,3670..4380)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(3709..3730)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3970..3989)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4317..4338
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4398..4874
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4597..4616)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4914..5054
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4914..5043
                     /label=AAV2 ITR
     rep_origin      5129..5584
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5216..5235)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5426..5447
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5601..5620)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5720..5742
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5780..5798)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5866..5970
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5971..6039,6040..6831)
                     /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(6189..6208)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7002..7590
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7491..7510
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtctagtt attaatagta atcaattacg gggtcattag
      181 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
      241 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
      301 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
      361 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
      421 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
      481 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
      541 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
      601 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
      661 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctcgtttag
      721 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagacacc
      781 gggaccgatc cagcctccgc ggattcgaat cccggccggg aacggtgcat tggaacgcgg
      841 attccccgtg ccaagagtga cgtaagtacc gcctatagag tctataggcc cacaaaaaat
      901 gctttcttct tttaatatac ttttttgttt atcttatttc taatactttc cctaatctct
      961 ttctttcagg gcaataatga tacaatgtat catgcctctt tgcaccattc taaagaataa
     1021 cagtgataat ttctgggtta aggcaatagc aatatttctg catataaata tttctgcata
     1081 taaattgtaa ctgatgtaag aggtttcata ttgctaatag cagctacaat ccagctacca
     1141 ttctgctttt attttatggt tgggataagg ctggattatt ctgagtccaa gctaggccct
     1201 tttgctaatc atgttcatac ctcttatctt cctcccacag ctcctgggca acgtgctggt
     1261 ctgtgtgctg gcccatcact ttggcaaaga attgggattc gaacatcgat tgaattcatg
     1321 catacaggag gagagacttc agcatgcaaa ccttcatctg tccggcttgc accgtcgttc
     1381 tcattccatg ctgctggcct tcagatggct gcacagatgc cccactcaca ccagtacagt
     1441 gaccgtcgcc agccgagcat aagtgaccag caggtgtctg ccttaccata ttctgaccag
     1501 attcagcaac ctctaactaa ccagaggcgg atgccccaaa ccttccgtga tccagcaact
     1561 gctcctctga gaaaactctc tgtggacttg atcaaaacat acaagcatat taatgaggtt
     1621 tactatgcaa aaaagaagcg aagacaccaa cagggccagg gggacgattc cagtcataag
     1681 aaggagcgga aggtttacaa tgatggttac gatgatgata actatgatta tattgtaaaa
     1741 aacggggaaa agtggatgga tcggtatgaa atcgactcct taataggcaa aggttcattt
     1801 ggacaggttg tgaaagctta tgacagagtg gagcaagaat gggtcgccat taaaatcatc
     1861 aagaacaaga aagcgtttct gaatcaagcc cagatagaag tgcggctgct tgagctcatg
     1921 aacaaacacg acactgaaat gaagtactac atagtgcatt tgaaacgcca ctttatgttt
     1981 cgaaaccatc tctgtttagt gtttgaaatg ctgtcctata atctctatga tttgttgaga
     2041 aacaccaact tccgaggggt ctctttgaac ctaacacgaa agtttgcgca acagatgtgc
     2101 acagcattgc tttttcttgc gactccagaa cttagtatca ttcactgtga cttaaagcct
     2161 gagaacatcc ttctgtgtaa ccccaaacgg agtgcaatca agattgttga ttttggcagc
     2221 tcttgtcagt tggggcagag gatataccag tatattcaga gtcgctttta tcggtctcca
     2281 gaggtgctac tgggaatgcc ttatgacctt gctatcgaca tgtggtccct tggatgtatc
     2341 ttggttgaaa tgcacactgg agagcctctg ttcagtggtg ccaatgaggt cgatcagatg
     2401 aataaaatag tggaagtctt gggcatccca cctgctcata ttcttgacca agcaccgaaa
     2461 gcaagaaagt tctttgagaa gttgcccgat ggcacttgga gcttaaagaa gaccaaagat
     2521 ggaaaacggg agtacaaacc accaggaacc cgtaaacttc ataatattct tggagtagaa
     2581 acaggaggac ctggcgggcg gcgtgctggg gaatcgggtc atactgtagc tgactacttg
     2641 aagttcaaag acctcatttt aaggatgctt gattatgacc ccaaaactcg gattcaacct
     2701 tattatgccc tgcagcacag ttttttcaag aaaacagctg atgaaggtac caacacaagt
     2761 aacagtgtgt ctaccagccc tgcgatggag cagtctcagt cttcaggcac cacctccagc
     2821 acctcctcca gctcaggtgg atcctcggga acgagtaaca gtgggagagc caggtcggat
     2881 ccgacgcacc agcatcgaca cagcggtgga cacttcgctg ctgctgtcca ggccatggac
     2941 tgtgagacac acagtcccca ggtgcgccag cagtttccgg ctcctctggg atggtcaggc
     3001 actgaagctc ctacacaagt cactgttgaa actcatcctg ttcaagagac aacctttcat
     3061 gtagcccccc agcagaacgc attgcatcat caccatggaa acagttccca tcaccaccac
     3121 caccaccatc accaccacca ccaccatgga cagcaagcct tgggtaaccg gaccaggcca
     3181 agggtctaca attctccaac aaatagctcc tctacccagg attctatgga ggttggccac
     3241 agtcaccact ccatgacatc cctgtcttcc tcaacaactt cttcctcgac atcttcctcc
     3301 tctactggta atcaaggcaa tcaggcctat cagaaccgcc cagtggctgc taacaccttg
     3361 gactttggac agaatggagc tatggacgtt aatttgaccg tctactccaa tccccgccaa
     3421 gagactggca tagctggaca tccaacatac caattttctg ctaatacagg tcctgcacat
     3481 tacatgactg aaggacatct ggcgatgaga caaggggctg atagagaaga gtctcccatg
     3541 acaggagttt gtgtgcaaca gagtcctgta gctagctcga ccggtgaagg ccgcggcagc
     3601 ctgctgacct gcggcgatgt ggaagaaaac ccgggcccgg cgccgggcag cactagtcgg
     3661 aagcttgtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
     3721 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc
     3781 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
     3841 accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg
     3901 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
     3961 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
     4021 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     4081 cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga caagcagaag
     4141 aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc
     4201 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     4261 cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg
     4321 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag
     4381 taactcgaga gatctacggg tggcatccct gtgacccctc cccagtgcct ctcctggccc
     4441 tggaagttgc cactccagtg cccaccagcc ttgtcctaat aaaattaagt tgcatcattt
     4501 tgtctgacta ggtgtccttc tataatatta tggggtggag gggggtggta tggagcaagg
     4561 ggcaagttgg gaagacaacc tgtagggcct gcggggtcta ttgggaacca agctggagtg
     4621 cagtggcaca atcttggctc actgcaatct ccgcctcctg ggttcaagcg attctcctgc
     4681 ctcagcctcc cgagttgttg ggattccagg catgcatgac caggctcagc taatttttgt
     4741 ttttttggta gagacggggt ttcaccatat tggccaggct ggtctccaac tcctaatctc
     4801 aggtgatcta cccaccttgg cctcccaaat tgctgggatt acaggcgtga accactgctc
     4861 ccttccctgt ccttctgatt ttgtaggtaa ccacgtgcgg accgagcggc cgcaggaacc
     4921 cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg
     4981 accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg
     5041 cagctgcctg caggggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     5101 acaccgcata cgtcaaagca accatagtac gcgccctgta gcggcgcatt aagcgcggcg
     5161 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccttagc gcccgctcct
     5221 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     5281 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     5341 gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     5401 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     5461 tctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggt ctattggtta
     5521 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca
     5581 attttatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga
     5641 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     5701 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     5761 aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     5821 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     5881 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     5941 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     6001 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     6061 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     6121 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     6181 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     6241 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     6301 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     6361 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     6421 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     6481 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     6541 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     6601 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     6661 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     6721 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     6781 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     6841 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     6901 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     6961 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     7021 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     7081 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     7141 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     7201 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     7261 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     7321 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     7381 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     7441 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     7501 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     7561 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     7621 ctggcctttt gctggccttt tgctcacatg t
//
