LOCUS       sequence-229495-        7182 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mouse DYRK4 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 7182)
  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 7182)
  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-DYRK4-2A-GFP
FEATURES             Location/Qualifiers
     source          1..7182
                     /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             2912..2923
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             3117..3170
                     /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(3198..3200,3201..3914)
                     /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(3198..3200,3201..3911)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(3240..3261)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3501..3520)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3848..3869
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3929..4405
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4128..4147)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4445..4585
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4445..4574
                     /label=AAV2 ITR
     rep_origin      4660..5115
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4747..4766)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4957..4978
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5132..5151)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5251..5273
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5311..5329)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5397..5501
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5502..5570,5571..6362)
                     /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(5720..5739)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6533..7121
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7022..7041
                     /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 tgaattcccc
     1321 ggggatccat gaggacgtcc aaggctcagt cggaggttga gagtaagcct ttgggtcacc
     1381 tccagaagcc atcctccaag aacaagaagc tgagagtgaa ccgatgtcca caaaagattc
     1441 caagtaacac ggcgttccct tttgtggaca caaaggggaa gaggaacaca gttaacttcc
     1501 cccagattgg caacaaagtc ccatcaaagg ggcccatgca gtatcaggaa aatcaaattc
     1561 ataatcaagt gctatcctct gaactcaaga cttcagaaat acctttcaac attaacacca
     1621 aagcccaaga caccaagcca cacccagaac tacaaaagaa gcacaaagtc cctcttacag
     1681 tagcagaggc cctaaagttc tttaagaatc agctgtcccc ttatgagcaa agcgaaatcc
     1741 tgggctactc agagctgtgg ttcctgggcc ttgaagccaa gaagctcaac gtggttccag
     1801 agaagttcag caagacaagc tttgatgatg agcacggttc ctatatgaag gtcctgcatg
     1861 accacattgc ctaccgatat gaggttctgg agatgatcgg gaaagggtcc tttggacagg
     1921 tggccaagtg cttggatcac aaaaacaatg agttggtggc cctgaaaatc atcaggaaca
     1981 aaaagagatt tcaccatcag gctctggtgg agctgaagat cctggaagcc ctcagaagga
     2041 aggacaaaga caacaaccac aatgtggtcc acatgaagga tttcttctac ttccggaatc
     2101 acctctgtat cacctttgag ctcctgggaa tcaacctgta tgagttgatg aagaacaaca
     2161 gctttcatgg cttcaatctg tccatagttc ggcgcttcac cttttccatt ttgaaatgct
     2221 tacacatgct ttatgtagag aaaatcatcc actgtgacct caagccggaa aatattgtgc
     2281 tataccagag gggccaagtc actgttaaag ttattgactt tgggtcaagc tgttatgaac
     2341 accagaaagt gtatacgtac atccaaagtc gtttctaccg atccccagag gtgattctgg
     2401 gacaccccta taacatggcc attgacatgt ggagcctggg ctgcatcatg gcggagctgt
     2461 acaccggata cccactgttc cctggggaga acgaggtgga gcagctggct tgcatcatgg
     2521 aggtgctggg cctgccacct gcccacttca cccagacagc ctccaggaga caggtattct
     2581 ttgattccaa aggtcttccc aaaaatataa acaacaacag agggggaaaa agatacccag
     2641 attccaagga tctcacgatg gtggtgaaaa cctacgattc cagcttcctg gactttctca
     2701 gaaggtggga gccttctctg cgcatgaccc ccgagcaggc cctcaagcat gcctggattc
     2761 acgagccgag gaaattcaaa ccacggccca agccacagat attaaggaag cctggtgcca
     2821 gtatttcctc tgagataagc acggaaaagg ctgaagagca acaggccagc aaaggcaaaa
     2881 aagatgaggc taccaaagaa actacagaca aattgaagga cgaagctgag aagcaccttg
     2941 agaattcggg caaacagcag agctctgtgg agcacacagc agacaccatc cagctgcctc
     3001 acctaacaga agcttctgga aagtcagaga ctgttgctgg gtcagagatg tccgcagaac
     3061 agcagagcac ctcctccccc aagagcacaa acatcttacc acccatcgtg accggtgaag
     3121 gccgcggcag cctgctgacc tgcggcgatg tggaagaaaa cccgggcccg gcgccgggca
     3181 gcactagtcg gaagcttgtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc
     3241 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg
     3301 gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg
     3361 tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc
     3421 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg
     3481 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg
     3541 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
     3601 acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg
     3661 acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca
     3721 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
     3781 tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc
     3841 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg
     3901 agctgtacaa gtaactcgag agatctacgg gtggcatccc tgtgacccct ccccagtgcc
     3961 tctcctggcc ctggaagttg ccactccagt gcccaccagc cttgtcctaa taaaattaag
     4021 ttgcatcatt ttgtctgact aggtgtcctt ctataatatt atggggtgga ggggggtggt
     4081 atggagcaag gggcaagttg ggaagacaac ctgtagggcc tgcggggtct attgggaacc
     4141 aagctggagt gcagtggcac aatcttggct cactgcaatc tccgcctcct gggttcaagc
     4201 gattctcctg cctcagcctc ccgagttgtt gggattccag gcatgcatga ccaggctcag
     4261 ctaatttttg tttttttggt agagacgggg tttcaccata ttggccaggc tggtctccaa
     4321 ctcctaatct caggtgatct acccaccttg gcctcccaaa ttgctgggat tacaggcgtg
     4381 aaccactgct cccttccctg tccttctgat tttgtaggta accacgtgcg gaccgagcgg
     4441 ccgcaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact
     4501 gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc
     4561 gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt attttctcct tacgcatctg
     4621 tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt agcggcgcat
     4681 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccttag
     4741 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     4801 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     4861 ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     4921 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     4981 caacactcaa ctctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg
     5041 tctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     5101 taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     5161 gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     5221 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     5281 cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta
     5341 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     5401 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     5461 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     5521 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     5581 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     5641 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     5701 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     5761 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     5821 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     5881 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     5941 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     6001 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     6061 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     6121 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     6181 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     6241 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     6301 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     6361 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     6421 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     6481 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     6541 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     6601 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     6661 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     6721 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     6781 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     6841 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     6901 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     6961 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     7021 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     7081 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     7141 ttttacggtt cctggccttt tgctggcctt ttgctcacat gt
//
