LOCUS Exported 7656 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses mouse DYRK1A tagged with 2A peptide at it's C-terminus and mCherry in mammalian cells.. ACCESSION . VERSION . KEYWORDS pAAV-DYRK1A-2A-mCherry 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 May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7656 /organism="synthetic DNA construct" /mol_type="other DNA" 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" 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 5976..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 //