LOCUS       sequence-229468-        7173 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mouse DYRK4 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 7173)
  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 7173)
  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-mCherry
FEATURES             Location/Qualifiers
     source          1..7173
                     /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             3198..3905
                     /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(3340..3358)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3608..3627
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3920..4396
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4119..4138)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4436..4576
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4436..4565
                     /label=AAV2 ITR
     rep_origin      4651..5106
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4738..4757)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4948..4969
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5123..5142)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5242..5264
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5302..5320)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5388..5492
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5493..5561,5562..6353)
                     /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(5711..5730)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6524..7112
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7013..7032
                     /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 aggaggataa catggccatc atcaaggagt
     3241 tcatgcgctt caaggtgcac atggagggct ccgtgaacgg ccacgagttc gagatcgagg
     3301 gcgagggcga gggccgcccc tacgagggca cccagaccgc caagctgaag gtgaccaagg
     3361 gtggccccct gcccttcgcc tgggacatcc tgtcccctca gttcatgtac ggctccaagg
     3421 cctacgtgaa gcaccccgcc gacatccccg actacttgaa gctgtccttc cccgagggct
     3481 tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg acccaggact
     3541 cctccctgca ggacggcgag ttcatctaca aggtgaagct gcgcggcacc aacttcccct
     3601 ccgacggccc cgtaatgcag aagaagacca tgggctggga ggcctcctcc gagcggatgt
     3661 accccgagga cggcgccctg aagggcgaga tcaagcagag gctgaagctg aaggacggcg
     3721 gccactacga cgctgaggtc aagaccacct acaaggccaa gaagcccgtg cagctgcccg
     3781 gcgcctacaa cgtcaacatc aagttggaca tcacctccca caacgaggac tacaccatcg
     3841 tggaacagta cgaacgcgcc gagggccgcc actccaccgg cggcatggac gagctgtaca
     3901 agtaactcga gagatctacg ggtggcatcc ctgtgacccc tccccagtgc ctctcctggc
     3961 cctggaagtt gccactccag tgcccaccag ccttgtccta ataaaattaa gttgcatcat
     4021 tttgtctgac taggtgtcct tctataatat tatggggtgg aggggggtgg tatggagcaa
     4081 ggggcaagtt gggaagacaa cctgtagggc ctgcggggtc tattgggaac caagctggag
     4141 tgcagtggca caatcttggc tcactgcaat ctccgcctcc tgggttcaag cgattctcct
     4201 gcctcagcct cccgagttgt tgggattcca ggcatgcatg accaggctca gctaattttt
     4261 gtttttttgg tagagacggg gtttcaccat attggccagg ctggtctcca actcctaatc
     4321 tcaggtgatc tacccacctt ggcctcccaa attgctggga ttacaggcgt gaaccactgc
     4381 tcccttccct gtccttctga ttttgtaggt aaccacgtgc ggaccgagcg gccgcaggaa
     4441 cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg
     4501 cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg
     4561 cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
     4621 tcacaccgca tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg
     4681 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc
     4741 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     4801 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     4861 ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     4921 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     4981 actctatctc gggctattct tttgatttat aagggatttt gccgatttcg gtctattggt
     5041 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta
     5101 caattttatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc
     5161 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     5221 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     5281 cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
     5341 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     5401 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     5461 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     5521 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     5581 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     5641 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     5701 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     5761 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     5821 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     5881 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     5941 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     6001 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     6061 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     6121 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     6181 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     6241 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     6301 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     6361 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     6421 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     6481 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     6541 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     6601 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     6661 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     6721 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     6781 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     6841 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     6901 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     6961 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     7021 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     7081 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     7141 tcctggcctt ttgctggcct tttgctcaca tgt
//
