LOCUS Exported 7651 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses mouse DYRK1A tagged with 2A peptide at it's C-terminus and GFP in mammalian cells.. ACCESSION . VERSION . KEYWORDS pAAV-DYRK1A-2A-GFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7651) 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 7651) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7651 /organism="synthetic DNA construct" /mol_type="other DNA" 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" 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..4383 /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 3667..4380 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE FVTAAGITLGMDELYK" primer_bind complement(3709..3730) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(3970..3989) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 4317..4338 /label=EGFP-C /note="EGFP, forward primer" polyA_signal 4398..4874 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(4597..4616) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" repeat_region 4914..5054 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 4914..5043 /label=AAV2 ITR rep_origin 5129..5584 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5216..5235) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 5426..5447 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(5601..5620) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 5720..5742 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(5780..5798) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 5866..5970 /gene="bla" /label=AmpR promoter CDS 5971..6831 /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(6189..6208) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 7002..7590 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 7491..7510 /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 ggagctgttc accggggtgg tgcccatcct ggtcgagctg 3721 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc 3781 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc 3841 accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg 3901 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc 3961 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc 4021 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg 4081 cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga caagcagaag 4141 aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc 4201 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac 4261 cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg 4321 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag 4381 taactcgaga gatctacggg tggcatccct gtgacccctc cccagtgcct ctcctggccc 4441 tggaagttgc cactccagtg cccaccagcc ttgtcctaat aaaattaagt tgcatcattt 4501 tgtctgacta ggtgtccttc tataatatta tggggtggag gggggtggta tggagcaagg 4561 ggcaagttgg gaagacaacc tgtagggcct gcggggtcta ttgggaacca agctggagtg 4621 cagtggcaca atcttggctc actgcaatct ccgcctcctg ggttcaagcg attctcctgc 4681 ctcagcctcc cgagttgttg ggattccagg catgcatgac caggctcagc taatttttgt 4741 ttttttggta gagacggggt ttcaccatat tggccaggct ggtctccaac tcctaatctc 4801 aggtgatcta cccaccttgg cctcccaaat tgctgggatt acaggcgtga accactgctc 4861 ccttccctgt ccttctgatt ttgtaggtaa ccacgtgcgg accgagcggc cgcaggaacc 4921 cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg 4981 accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg 5041 cagctgcctg caggggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc 5101 acaccgcata cgtcaaagca accatagtac gcgccctgta gcggcgcatt aagcgcggcg 5161 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccttagc gcccgctcct 5221 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat 5281 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt 5341 gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg 5401 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac 5461 tctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggt ctattggtta 5521 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca 5581 attttatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga 5641 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac 5701 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg 5761 aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata 5821 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt 5881 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa 5941 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt 6001 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa 6061 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac 6121 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt 6181 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt 6241 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat 6301 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac 6361 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg 6421 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc 6481 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa 6541 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag 6601 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct 6661 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat 6721 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa 6781 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac 6841 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc 6901 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc 6961 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg 7021 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg 7081 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca 7141 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg 7201 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg 7261 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga 7321 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac 7381 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat 7441 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc 7501 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga 7561 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc 7621 ctggcctttt gctggccttt tgctcacatg t //