LOCUS       sequence-223476-        7656 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 7656)
  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 7656)
  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..7656
                     /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             3123..3152
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             3126..3155
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             3129..3158
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             3132..3161
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             3600..3653
                     /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             3681..4388
                     /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(3823..3841)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4091..4110
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    4403..4879
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4602..4621)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4919..5059
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4919..5048
                     /label=AAV2 ITR
     rep_origin      5134..5589
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5221..5240)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5431..5452
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5606..5625)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5725..5747
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5785..5803)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5871..5975
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5976..6044,6045..6836)
                     /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(6194..6213)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7007..7595
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7496..7515
                     /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 catgcataca ggaggagaga cttcagcatg caaaccttca tctgtccggc
     1381 ttgcaccgtc gttctcattc catgctgctg gccttcagat ggctgcacag atgccccact
     1441 cacaccagta cagtgaccgt cgccagccga gcataagtga ccagcaggtg tctgccttac
     1501 catattctga ccagattcag caacctctaa ctaaccagag gcggatgccc caaaccttcc
     1561 gtgatccagc aactgctcct ctgagaaaac tctctgtgga cttgatcaaa acatacaagc
     1621 atattaatga ggtttactat gcaaaaaaga agcgaagaca ccaacagggc cagggggacg
     1681 attccagtca taagaaggag cggaaggttt acaatgatgg ttacgatgat gataactatg
     1741 attatattgt aaaaaacggg gaaaagtgga tggatcggta tgaaatcgac tccttaatag
     1801 gcaaaggttc atttggacag gttgtgaaag cttatgacag agtggagcaa gaatgggtcg
     1861 ccattaaaat catcaagaac aagaaagcgt ttctgaatca agcccagata gaagtgcggc
     1921 tgcttgagct catgaacaaa cacgacactg aaatgaagta ctacatagtg catttgaaac
     1981 gccactttat gtttcgaaac catctctgtt tagtgtttga aatgctgtcc tataatctct
     2041 atgatttgtt gagaaacacc aacttccgag gggtctcttt gaacctaaca cgaaagtttg
     2101 cgcaacagat gtgcacagca ttgctttttc ttgcgactcc agaacttagt atcattcact
     2161 gtgacttaaa gcctgagaac atccttctgt gtaaccccaa acggagtgca atcaagattg
     2221 ttgattttgg cagctcttgt cagttggggc agaggatata ccagtatatt cagagtcgct
     2281 tttatcggtc tccagaggtg ctactgggaa tgccttatga ccttgctatc gacatgtggt
     2341 cccttggatg tatcttggtt gaaatgcaca ctggagagcc tctgttcagt ggtgccaatg
     2401 aggtcgatca gatgaataaa atagtggaag tcttgggcat cccacctgct catattcttg
     2461 accaagcacc gaaagcaaga aagttctttg agaagttgcc cgatggcact tggagcttaa
     2521 agaagaccaa agatggaaaa cgggagtaca aaccaccagg aacccgtaaa cttcataata
     2581 ttcttggagt agaaacagga ggacctggcg ggcggcgtgc tggggaatcg ggtcatactg
     2641 tagctgacta cttgaagttc aaagacctca ttttaaggat gcttgattat gaccccaaaa
     2701 ctcggattca accttattat gccctgcagc acagtttttt caagaaaaca gctgatgaag
     2761 gtaccaacac aagtaacagt gtgtctacca gccctgcgat ggagcagtct cagtcttcag
     2821 gcaccacctc cagcacctcc tccagctcag gtggatcctc gggaacgagt aacagtggga
     2881 gagccaggtc ggatccgacg caccagcatc gacacagcgg tggacacttc gctgctgctg
     2941 tccaggccat ggactgtgag acacacagtc cccaggtgcg ccagcagttt ccggctcctc
     3001 tgggatggtc aggcactgaa gctcctacac aagtcactgt tgaaactcat cctgttcaag
     3061 agacaacctt tcatgtagcc ccccagcaga acgcattgca tcatcaccat ggaaacagtt
     3121 cccatcacca ccaccaccac catcaccacc accaccacca tggacagcaa gccttgggta
     3181 accggaccag gccaagggtc tacaattctc caacaaatag ctcctctacc caggattcta
     3241 tggaggttgg ccacagtcac cactccatga catccctgtc ttcctcaaca acttcttcct
     3301 cgacatcttc ctcctctact ggtaatcaag gcaatcaggc ctatcagaac cgcccagtgg
     3361 ctgctaacac cttggacttt ggacagaatg gagctatgga cgttaatttg accgtctact
     3421 ccaatccccg ccaagagact ggcatagctg gacatccaac ataccaattt tctgctaata
     3481 caggtcctgc acattacatg actgaaggac atctggcgat gagacaaggg gctgatagag
     3541 aagagtctcc catgacagga gtttgtgtgc aacagagtcc tgtagctagc tcgaccggtg
     3601 aaggccgcgg cagcctgctg acctgcggcg atgtggaaga aaacccgggc ccggcgccgg
     3661 gcagcactag tcggaagctt gtgagcaagg gcgaggagga taacatggcc atcatcaagg
     3721 agttcatgcg cttcaaggtg cacatggagg gctccgtgaa cggccacgag ttcgagatcg
     3781 agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg aaggtgacca
     3841 agggtggccc cctgcccttc gcctgggaca tcctgtcccc tcagttcatg tacggctcca
     3901 aggcctacgt gaagcacccc gccgacatcc ccgactactt gaagctgtcc ttccccgagg
     3961 gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc gtgacccagg
     4021 actcctccct gcaggacggc gagttcatct acaaggtgaa gctgcgcggc accaacttcc
     4081 cctccgacgg ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc tccgagcgga
     4141 tgtaccccga ggacggcgcc ctgaagggcg agatcaagca gaggctgaag ctgaaggacg
     4201 gcggccacta cgacgctgag gtcaagacca cctacaaggc caagaagccc gtgcagctgc
     4261 ccggcgccta caacgtcaac atcaagttgg acatcacctc ccacaacgag gactacacca
     4321 tcgtggaaca gtacgaacgc gccgagggcc gccactccac cggcggcatg gacgagctgt
     4381 acaagtaact cgagagatct acgggtggca tccctgtgac ccctccccag tgcctctcct
     4441 ggccctggaa gttgccactc cagtgcccac cagccttgtc ctaataaaat taagttgcat
     4501 cattttgtct gactaggtgt ccttctataa tattatgggg tggagggggg tggtatggag
     4561 caaggggcaa gttgggaaga caacctgtag ggcctgcggg gtctattggg aaccaagctg
     4621 gagtgcagtg gcacaatctt ggctcactgc aatctccgcc tcctgggttc aagcgattct
     4681 cctgcctcag cctcccgagt tgttgggatt ccaggcatgc atgaccaggc tcagctaatt
     4741 tttgtttttt tggtagagac ggggtttcac catattggcc aggctggtct ccaactccta
     4801 atctcaggtg atctacccac cttggcctcc caaattgctg ggattacagg cgtgaaccac
     4861 tgctcccttc cctgtccttc tgattttgta ggtaaccacg tgcggaccga gcggccgcag
     4921 gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc
     4981 gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga
     5041 gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc tccttacgca tctgtgcggt
     5101 atttcacacc gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg
     5161 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     5221 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     5281 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     5341 aacttgattt gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     5401 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     5461 tcaaccctat ctcgggctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     5521 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt
     5581 ttacaatttt atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc
     5641 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
     5701 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
     5761 caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca
     5821 tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     5881 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     5941 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     6001 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     6061 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     6121 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     6181 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     6241 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     6301 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     6361 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     6421 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     6481 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     6541 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     6601 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     6661 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     6721 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     6781 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     6841 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     6901 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     6961 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     7021 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     7081 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     7141 taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     7201 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     7261 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     7321 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     7381 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     7441 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     7501 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     7561 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     7621 ggttcctggc cttttgctgg ccttttgctc acatgt
//
