LOCUS       sequence-229497-        7163 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mouse DYRK3 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 7163)
  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 7163)
  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-DYRK3-2A-GFP
FEATURES             Location/Qualifiers
     source          1..7163
                     /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             3098..3151
                     /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(3179..3181,3182..3895)
                     /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(3179..3181,3182..3892)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(3221..3242)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3482..3501)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3829..3850
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3910..4386
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4109..4128)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4426..4566
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4426..4555
                     /label=AAV2 ITR
     rep_origin      4641..5096
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4728..4747)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4938..4959
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5113..5132)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5232..5254
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5292..5310)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5378..5482
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5483..5551,5552..6343)
                     /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(5701..5720)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6514..7102
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7003..7022
                     /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 tgaattcaat
     1321 gggaggcgca gcccgcgatc gcgggaggaa ggacgcggcg ctgccggggg ccgggctccc
     1381 gccgcagcag cggaggttgg gggatggtgt ctatgatact ttcatgatga tagatgaaac
     1441 caagggccca ccctattcgg acacattcag caacccctct gaagcacctg tctccagaag
     1501 gctaaatatt accactgagc cactcacgag aggccacact cagcactttg tgaatggaag
     1561 tgagatgaag gtggaacaac tgtttcaaga atttggcaac agaagatcca atactctcca
     1621 gtcagatggc atcagcaact cggaaaagtc ttctccggct tctcagggga aaagttcaga
     1681 aagcctgagc gcggtgaagt gcaacctttc ctccaggccg tctaaggtgc tgccgctgac
     1741 tcctgagcaa gcgctgaagc agtataaaca ccacctcact gcctacgaga agctggagat
     1801 cgtcagctac ccagaaatct actttgtggg tccgaatgcc aaaaagcggc agggagttat
     1861 tggtggtccc aataacggtg gctacgacga tgcggatggg gcctatattc atgtgccccg
     1921 ggaccatctg gcttaccgct atgaggtgct gaaaatcatc ggcaagggga gttttggaca
     1981 ggtagcccgg gtctatgatc acaaacttcg gcagtacgtg gccctgaaaa tggtgcgcaa
     2041 tgagaaacgc ttccatcgcc aggcagccga ggagatccgg atcttggagc atcttaaaaa
     2101 gcaagacaaa actggtagca tgaacgtcat ccacatgcta gaaagtttca ccttccggaa
     2161 ccacgtgtgc atggcctttg aactgctaag catagacctg tatgagctta ttaaaaaaaa
     2221 caagttccag ggcttcagcg tccagttggt tcggaagttc gcccagtcca tcctacagtc
     2281 cctggacgct ctgcacaaaa acaagatcat ccactgcgac ctgaagccgg aaaacattct
     2341 cctgaaacac cacggccgaa gcgccaccaa ggtcatcgac tttggctcca gctgcttcga
     2401 gtatcagaag ctttacacgt atatccagtc ccgcttctac agagccccag agatcatcct
     2461 ggggtgccgc tacagcaccc cgatcgacat atggagcttt ggttgcatcc ttgcagaact
     2521 tttgacagga cagcccctgt tccctggaga ggacgaagga gaccagttgg cctgcatgat
     2581 agagttgcta ggaatgccac cgcagaaact tctggagcaa tccaagcgtg ccaagtactt
     2641 tattaactcc aaaggcttgc ctcgatactg ctccgtatct acccagacgg acgggagggt
     2701 ggtgcttctc gggggtcgct cacgcagggg taaaaagcga ggcccgccag gcagcaaaga
     2761 ctgggcaacc gcactgaagg gctgtggtga ctacttgttc atagagtttc tgaaacgatg
     2821 cctccagtgg gacccctctg cccgcctcac cccggctcaa gcattaagac atccttggat
     2881 tagcaagtct acacccaaac ctctcaccat ggacaaggtg ccagggaagc gggtagttaa
     2941 ccctacaaat gctttccagg gactgggttc caagctgcct ccagtcgttg ggatagccag
     3001 taagcttaaa gctaacctaa tgtccgaaac cagtggtagt atacctctgt gcagtgtatt
     3061 gccaaagctg attagcctcg accggggtac caccggtgaa ggccgcggca gcctgctgac
     3121 ctgcggcgat gtggaagaaa acccgggccc ggcgccgggc agcactagtc ggaagcttgt
     3181 gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga
     3241 cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa
     3301 gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt
     3361 gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca
     3421 cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa
     3481 ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa
     3541 ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct
     3601 ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat
     3661 caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca
     3721 ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct
     3781 gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct
     3841 ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca agtaactcga
     3901 gagatctacg ggtggcatcc ctgtgacccc tccccagtgc ctctcctggc cctggaagtt
     3961 gccactccag tgcccaccag ccttgtccta ataaaattaa gttgcatcat tttgtctgac
     4021 taggtgtcct tctataatat tatggggtgg aggggggtgg tatggagcaa ggggcaagtt
     4081 gggaagacaa cctgtagggc ctgcggggtc tattgggaac caagctggag tgcagtggca
     4141 caatcttggc tcactgcaat ctccgcctcc tgggttcaag cgattctcct gcctcagcct
     4201 cccgagttgt tgggattcca ggcatgcatg accaggctca gctaattttt gtttttttgg
     4261 tagagacggg gtttcaccat attggccagg ctggtctcca actcctaatc tcaggtgatc
     4321 tacccacctt ggcctcccaa attgctggga ttacaggcgt gaaccactgc tcccttccct
     4381 gtccttctga ttttgtaggt aaccacgtgc ggaccgagcg gccgcaggaa cccctagtga
     4441 tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg
     4501 tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc
     4561 tgcaggggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     4621 tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     4681 ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt
     4741 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     4801 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg
     4861 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     4921 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca actctatctc
     4981 gggctattct tttgatttat aagggatttt gccgatttcg gtctattggt taaaaaatga
     5041 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg
     5101 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc
     5161 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     5221 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     5281 gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     5341 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     5401 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     5461 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     5521 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     5581 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     5641 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     5701 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     5761 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     5821 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     5881 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     5941 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     6001 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     6061 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     6121 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     6181 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     6241 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     6301 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     6361 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     6421 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     6481 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     6541 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     6601 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     6661 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     6721 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     6781 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     6841 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     6901 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     6961 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     7021 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     7081 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     7141 ttgctggcct tttgctcaca tgt
//
