LOCUS       sequence-223498-        7177 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 7177)
  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 7177)
  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..7177
                     /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             3112..3165
                     /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(3193..3195,3196..3909)
                     /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(3193..3195,3196..3906)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(3235..3256)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3496..3515)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3843..3864
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3924..4400
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4123..4142)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4440..4580
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4440..4569
                     /label=AAV2 ITR
     rep_origin      4655..5110
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4742..4761)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4952..4973
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5127..5146)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5246..5268
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5306..5324)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5392..5496
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5497..5565,5566..6357)
                     /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(5715..5734)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6528..7116
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7017..7036
                     /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 caatgggagg cgcagcccgc gatcgcggga ggaaggacgc ggcgctgccg
     1381 ggggccgggc tcccgccgca gcagcggagg ttgggggatg gtgtctatga tactttcatg
     1441 atgatagatg aaaccaaggg cccaccctat tcggacacat tcagcaaccc ctctgaagca
     1501 cctgtctcca gaaggctaaa tattaccact gagccactca cgagaggcca cactcagcac
     1561 tttgtgaatg gaagtgagat gaaggtggaa cagctgtttc aagaatttgg caacagaaga
     1621 tccaatactc tccagtcaga tggcatcagc aactcggaaa agtcttctcc ggcttctcag
     1681 gggaaaagtt cggaaagcct gagcgcggtg aagtgcaacc tttcctccag gccgtctaag
     1741 gtgctgccgc tgactcctga gcaagcgctg aagcagtata aacaccacct cactgcctac
     1801 gagaagctgg agatcgtcag ctacccagaa atctactttg tgggtccgaa tgccaaaaag
     1861 cggcagggag ttattggtgg tcccaataac ggtggctacg acgatgcgga tggggcctat
     1921 attcatgtgc cccgggacca tctggcttac cgctatgagg tgctgaaaat catcggcaag
     1981 gggagttttg gacaggtagc ccgggtctat gatcacaaac ttcggcagta cgtggccctg
     2041 aaaatggtgc gcaatgagaa acgcttccat cgccaggcgg ccgaggagat ccggatcttg
     2101 gagcatctta aaaagcaaga caaaactggt agcatgaacg tcatccacat gctagaaagt
     2161 ttcaccttcc ggaaccacgt gtgcatggcc tttgaactgc taagcataga cctgtatgag
     2221 cttattaaaa aaaacaagtt ccagggcttc agcgtccagt tggttcggaa gttcgcccag
     2281 tccatcctac agtccctgga cgctctgcac aaaaacaaga tcatccactg cgacctgaag
     2341 ccggaaaaca ttctcctgaa acaccacggc cgaagcgcca ccaaggtcat cgactttggc
     2401 tccagctgct tcgagtatca gaagctttac acgtatatcc agtcccgctt ctacagagcc
     2461 ccagagatca tcctggggtg ccgctacagc accccgatcg acatatggag ctttggttgc
     2521 atccttgcag aacttttgac aggacagccc ctgttccctg gagaggacga aggagaccag
     2581 ttggcctgca tgatagagtt gctaggaatg ccaccgcaga aacttctgga gcaatccaag
     2641 cgtgccaagt actttattaa ctccaaaggc ttgcctcgat actgctccgt atctacccag
     2701 acggacggga gggtggtgct tctcgggggt cgctcacgca ggggtaaaaa gcgaggcccg
     2761 ccaggcagca aagactgggc aaccgcactg aagggctgtg gtgactactt gttcatagag
     2821 tttctgaaac gatgcctcca gtgggacccc tctgcccgcc tcaccccggc tcaagcatta
     2881 agacatcctt ggattagcaa gtctacaccc aaacctctca ccatggacaa ggtgccaggg
     2941 aagcgggtag ttaaccctac aaatgctttc cagggactgg gttccaagct gcctccagtc
     3001 gttgggatag ccagtaagct taaagctaac ctaatgtccg aaaccagtgg tagtatacct
     3061 ctgtgcagtg tattgccaaa gctgattagc ctcgaccggg gtaccaccgg tgaaggccgc
     3121 ggcagcctgc tgacctgcgg cgatgtggaa gaaaacccgg gcccggcgcc gggcagcact
     3181 agtcggaagc ttgtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
     3241 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
     3301 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
     3361 tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac
     3421 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
     3481 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
     3541 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     3601 ctggggcaca agctggagta caactacaac agccacaacg tctatatcat ggccgacaag
     3661 cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg
     3721 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     3781 gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat
     3841 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     3901 tacaagtaac tcgagagatc tacgggtggc atccctgtga cccctcccca gtgcctctcc
     3961 tggccctgga agttgccact ccagtgccca ccagccttgt cctaataaaa ttaagttgca
     4021 tcattttgtc tgactaggtg tccttctata atattatggg gtggaggggg gtggtatgga
     4081 gcaaggggca agttgggaag acaacctgta gggcctgcgg ggtctattgg gaaccaagct
     4141 ggagtgcagt ggcacaatct tggctcactg caatctccgc ctcctgggtt caagcgattc
     4201 tcctgcctca gcctcccgag ttgttgggat tccaggcatg catgaccagg ctcagctaat
     4261 ttttgttttt ttggtagaga cggggtttca ccatattggc caggctggtc tccaactcct
     4321 aatctcaggt gatctaccca ccttggcctc ccaaattgct gggattacag gcgtgaacca
     4381 ctgctccctt ccctgtcctt ctgattttgt aggtaaccac gtgcggaccg agcggccgca
     4441 ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc
     4501 cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg
     4561 agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg
     4621 tatttcacac cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc
     4681 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     4741 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     4801 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     4861 aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     4921 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     4981 ctcaacccta tctcgggcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     5041 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     5101 tttacaattt tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     5161 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
     5221 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
     5281 tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
     5341 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     5401 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     5461 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     5521 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     5581 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     5641 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     5701 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     5761 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     5821 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     5881 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     5941 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     6001 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     6061 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     6121 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     6181 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     6241 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     6301 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     6361 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     6421 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     6481 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     6541 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     6601 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     6661 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     6721 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     6781 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     6841 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     6901 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     6961 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     7021 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     7081 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     7141 cggttcctgg ccttttgctg gccttttgct cacatgt
//
