LOCUS       sequence-223589-        7187 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 7187)
  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 7187)
  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..7187
                     /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             2926..2937
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             3131..3184
                     /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             3212..3919
                     /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(3354..3372)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3622..3641
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3934..4410
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4133..4152)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4450..4590
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4450..4579
                     /label=AAV2 ITR
     rep_origin      4665..5120
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4752..4771)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4962..4983
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5137..5156)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5256..5278
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5316..5334)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5402..5506
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5507..5575,5576..6367)
                     /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(5725..5744)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6538..7126
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7027..7046
                     /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 ccccggggat ccatgaggac gtccaaggct cagtcggagg ttgagagtaa
     1381 gcctttgggt cacctccaga agccatcctc caagaacaag aagctgagag tgaaccgatg
     1441 tccacaaaag attccaagta acacggcgtt cccttttgtg gacacaaagg ggaagaggaa
     1501 cacagttaac ttcccccaga ttggcaacaa agtcccatca aaggggccca tgcagtatca
     1561 ggaaaatcaa attcataatc aagtgctatc ctctgaactc aagacttcag aaataccttt
     1621 caacattaac accaaagccc aagacaccaa gccacaccca gaactacaaa agaagcacaa
     1681 agtccctctt acagtagcag aggccctaaa gttctttaag aatcagctgt ccccttatga
     1741 gcaaagcgaa atcctgggct actcagagct gtggttcctg ggccttgaag ccaagaagct
     1801 caacgtggtt ccagagaagt tcagcaagac aagctttgat gatgagcacg gttcctatat
     1861 gaaggtcctg catgaccaca ttgcctaccg atatgaggtt ctggagatga tcgggaaagg
     1921 gtcctttgga caggtggcca agtgcttgga tcacaaaaac aatgagttgg tggccctgaa
     1981 aatcatcagg aacaaaaaga gatttcacca tcaggctctg gtggagctga agatcctgga
     2041 agccctcaga aggaaggaca aagacaacaa ccacaatgtg gtccacatga aggatttctt
     2101 ctacttccgg aatcacctct gtatcacctt tgagctcctg ggaatcaacc tgtatgagtt
     2161 gatgaagaac aacagctttc atggcttcaa tctgtccata gttcggcgct tcaccttttc
     2221 cattttgaaa tgcttacaca tgctttatgt agagaaaatc atccactgtg acctcaagcc
     2281 ggaaaatatt gtgctatacc agaggggcca agtcactgtt aaagttattg actttgggtc
     2341 aagctgttat gaacaccaga aagtgtatac gtacatccaa agtcgtttct accgatcccc
     2401 agaggtgatt ctgggacacc cctataacat ggccattgac atgtggagcc tgggctgcat
     2461 catggcggag ctgtacaccg gatacccact gttccctggg gagaacgagg tggagcagct
     2521 ggcttgcatc atggaggtgc tgggcctgcc acctgcccac ttcacccaga cagcctccag
     2581 gagacaggta ttctttgatt ccaaaggtct tcccaaaaat ataaacaaca acagaggggg
     2641 aaaaagatac ccagattcca aggatctcac gatggtggtg aaaacctacg attccagctt
     2701 cctggacttt ctcagaaggt gggagccttc tctgcgcatg acccccgagc aggccctcaa
     2761 gcatgcctgg attcacgagc cgaggaaatt caaaccacgg cccaagccac agatattaag
     2821 gaagcctggt gccagtattt cctctgagat aagcacggaa aaggctgaag agcaacaggc
     2881 cagcaaaggc aaaaaagatg aggctaccaa agaaactaca gacaaattga aggacgaagc
     2941 tgagaagcac cttgagaatt cgggcaaaca gcagagctct gtggagcaca cagcagacac
     3001 catccagctg cctcacctaa cagaagcttc tggaaagtca gagactgttg ctgggtcaga
     3061 gatgtccgca gaacagcaga gcacctcctc ccccaagagc acaaacatct taccacccat
     3121 cgtgaccggt gaaggccgcg gcagcctgct gacctgcggc gatgtggaag aaaacccggg
     3181 cccggcgccg ggcagcacta gtcggaagct tgtgagcaag ggcgaggagg ataacatggc
     3241 catcatcaag gagttcatgc gcttcaaggt gcacatggag ggctccgtga acggccacga
     3301 gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga ccgccaagct
     3361 gaaggtgacc aagggtggcc ccctgccctt cgcctgggac atcctgtccc ctcagttcat
     3421 gtacggctcc aaggcctacg tgaagcaccc cgccgacatc cccgactact tgaagctgtc
     3481 cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac
     3541 cgtgacccag gactcctccc tgcaggacgg cgagttcatc tacaaggtga agctgcgcgg
     3601 caccaacttc ccctccgacg gccccgtaat gcagaagaag accatgggct gggaggcctc
     3661 ctccgagcgg atgtaccccg aggacggcgc cctgaagggc gagatcaagc agaggctgaa
     3721 gctgaaggac ggcggccact acgacgctga ggtcaagacc acctacaagg ccaagaagcc
     3781 cgtgcagctg cccggcgcct acaacgtcaa catcaagttg gacatcacct cccacaacga
     3841 ggactacacc atcgtggaac agtacgaacg cgccgagggc cgccactcca ccggcggcat
     3901 ggacgagctg tacaagtaac tcgagagatc tacgggtggc atccctgtga cccctcccca
     3961 gtgcctctcc tggccctgga agttgccact ccagtgccca ccagccttgt cctaataaaa
     4021 ttaagttgca tcattttgtc tgactaggtg tccttctata atattatggg gtggaggggg
     4081 gtggtatgga gcaaggggca agttgggaag acaacctgta gggcctgcgg ggtctattgg
     4141 gaaccaagct ggagtgcagt ggcacaatct tggctcactg caatctccgc ctcctgggtt
     4201 caagcgattc tcctgcctca gcctcccgag ttgttgggat tccaggcatg catgaccagg
     4261 ctcagctaat ttttgttttt ttggtagaga cggggtttca ccatattggc caggctggtc
     4321 tccaactcct aatctcaggt gatctaccca ccttggcctc ccaaattgct gggattacag
     4381 gcgtgaacca ctgctccctt ccctgtcctt ctgattttgt aggtaaccac gtgcggaccg
     4441 agcggccgca ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc
     4501 tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag
     4561 tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc
     4621 atctgtgcgg tatttcacac cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg
     4681 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     4741 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     4801 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     4861 cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac
     4921 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     4981 tggaacaaca ctcaacccta tctcgggcta ttcttttgat ttataaggga ttttgccgat
     5041 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     5101 aatattaacg tttacaattt tatggtgcac tctcagtaca atctgctctg atgccgcata
     5161 gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
     5221 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
     5281 ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata
     5341 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
     5401 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     5461 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     5521 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     5581 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     5641 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     5701 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     5761 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     5821 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     5881 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     5941 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     6001 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     6061 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     6121 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     6181 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     6241 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     6301 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     6361 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     6421 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     6481 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     6541 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     6601 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     6661 cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa
     6721 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     6781 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     6841 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     6901 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     6961 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     7021 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     7081 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     7141 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgt
//
