LOCUS       sequence-229471-        7642 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mouse DYRK1A tagged with 2A peptide at it's C-terminus and
            mCherry in mammalian cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7642)
  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 7642)
  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-mCherry
FEATURES             Location/Qualifiers
     source          1..7642
                     /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             3667..4374
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(3809..3827)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4077..4096
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    4389..4865
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4588..4607)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4905..5045
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4905..5034
                     /label=AAV2 ITR
     rep_origin      5120..5575
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5207..5226)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5417..5438
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5592..5611)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5711..5733
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5771..5789)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5857..5961
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5962..6030,6031..6822)
                     /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(6180..6199)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6993..7581
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7482..7501
                     /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 ggaggataac atggccatca tcaaggagtt catgcgcttc
     3721 aaggtgcaca tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgagggcgag
     3781 ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg tggccccctg
     3841 cccttcgcct gggacatcct gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag
     3901 caccccgccg acatccccga ctacttgaag ctgtccttcc ccgagggctt caagtgggag
     3961 cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag
     4021 gacggcgagt tcatctacaa ggtgaagctg cgcggcacca acttcccctc cgacggcccc
     4081 gtaatgcaga agaagaccat gggctgggag gcctcctccg agcggatgta ccccgaggac
     4141 ggcgccctga agggcgagat caagcagagg ctgaagctga aggacggcgg ccactacgac
     4201 gctgaggtca agaccaccta caaggccaag aagcccgtgc agctgcccgg cgcctacaac
     4261 gtcaacatca agttggacat cacctcccac aacgaggact acaccatcgt ggaacagtac
     4321 gaacgcgccg agggccgcca ctccaccggc ggcatggacg agctgtacaa gtaactcgag
     4381 agatctacgg gtggcatccc tgtgacccct ccccagtgcc tctcctggcc ctggaagttg
     4441 ccactccagt gcccaccagc cttgtcctaa taaaattaag ttgcatcatt ttgtctgact
     4501 aggtgtcctt ctataatatt atggggtgga ggggggtggt atggagcaag gggcaagttg
     4561 ggaagacaac ctgtagggcc tgcggggtct attgggaacc aagctggagt gcagtggcac
     4621 aatcttggct cactgcaatc tccgcctcct gggttcaagc gattctcctg cctcagcctc
     4681 ccgagttgtt gggattccag gcatgcatga ccaggctcag ctaatttttg tttttttggt
     4741 agagacgggg tttcaccata ttggccaggc tggtctccaa ctcctaatct caggtgatct
     4801 acccaccttg gcctcccaaa ttgctgggat tacaggcgtg aaccactgct cccttccctg
     4861 tccttctgat tttgtaggta accacgtgcg gaccgagcgg ccgcaggaac ccctagtgat
     4921 ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt
     4981 cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct
     5041 gcaggggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
     5101 acgtcaaagc aaccatagta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     5161 gttacgcgca gcgtgaccgc tacacttgcc agcgccttag cgcccgctcc tttcgctttc
     5221 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     5281 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgatttgggt
     5341 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     5401 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ctctatctcg
     5461 ggctattctt ttgatttata agggattttg ccgatttcgg tctattggtt aaaaaatgag
     5521 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aattttatgg
     5581 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca
     5641 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
     5701 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
     5761 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     5821 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     5881 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     5941 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     6001 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     6061 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     6121 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     6181 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     6241 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     6301 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     6361 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     6421 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     6481 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     6541 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     6601 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     6661 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     6721 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     6781 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     6841 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     6901 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     6961 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     7021 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     7081 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     7141 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     7201 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     7261 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     7321 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     7381 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     7441 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     7501 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     7561 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     7621 tgctggcctt ttgctcacat gt
//
