LOCUS Exported 5519 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION An AAV genome that expresses nuclear localized mRuby2 from the mouse Dlx5/6 enhancer. ACCESSION . VERSION . KEYWORDS pAAV-mDlx-NLS-mRuby2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5519) AUTHORS Chan KY, Jang MJ, Yoo BB, Greenbaum A, Ravi N, Wu WL, Sanchez-Guardado L, Lois C, Mazmanian SK, Deverman BE, Gradinaru V TITLE Engineered AAVs for efficient noninvasive gene delivery to the central and peripheral nervous systems. JOURNAL Nat Neurosci. 2017 Aug;20(8):1172-1179. doi: 10.1038/nn.4593. Epub 2017 Jun 26. PUBMED 28671695 REFERENCE 2 (bases 1 to 5519) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5519 /organism="synthetic DNA construct" /mol_type="other DNA" repeat_region 1..130 /label=AAV2 ITR (alternate) /note="Functional equivalent of wild-type AAV2 ITR" intron 849..981 /label=chimeric intron /note="chimera between introns from human beta-globin and immunoglobulin heavy chain genes" primer_bind 964..983 /label=pMT2-F /note="Synthetic intron, forward primer" CDS 1078..1098 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 1108..1818 /codon_start=1 /product="monomeric red fluorescent protein derived from mRuby, with improved photophysical properties (Lam et al., 2012)" /label=mRuby2 /translation="MVSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGNPYMGTQTM RIKVIEGGPLPFAFDILATSFMYGSRTFIKYPKGIPDFFKQSFPEGFTWERVTRYEDGG VVTVMQDTSLEDGCLVYHVQVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTH MALKVDGGGHLSCSFVTTYRSKKTVGNIKMPGIHAVDHRLERLEESDNEMFVVQREHAV AKFAGLGGGMDELYK" misc_feature 1865..2453 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(1918..1938) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(2336..2347) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 2622..2703 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2659..2678) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2713..2732 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" repeat_region 2782..2922 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 2782..2911 /label=AAV2 ITR rep_origin 2997..3452 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3084..3103) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3294..3315 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3469..3488) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 3588..3610 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(3648..3666) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 3734..3838 /gene="bla" /label=AmpR promoter CDS 3839..4699 /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(4057..4076) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 4870..5458 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5359..5378 /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 gcgtttaatt aagaattcta tacactcaca gtggtttggc 181 atatatttgg tgaaattttt taaggaaaaa ttagtgttgg tttcgatata tggtagcttt 241 ttctctaaca taatttgaat aattcagcaa agccctacta ccagctgtac ttctgcagcc 301 tcttccattc tttccagcat tataattttg gttaattttc aattttaggt cctacgtctc 361 tgcaatttgt gtatgaataa cagaataatt tccctctttt gtttcgcctt tcctgttcct 421 gaatctaaat aaagatggct ttttagtatt aaaagtggaa gaaaattaca ggtaattatc 481 tttgacggta aaaacgctgt aatcagcggg ctacatgaaa aattactcta attatggctg 541 catttaagag aatggaaaaa aaccttcttg tggataaaaa ccttaaattg tccccaatgt 601 ctgcttcaaa ttggatggca ctgcagctgg aggctttgtt cagaattgat cctggggagc 661 tacgaaccca aagtttcaca gtaggaagct tgatatcgaa ttcctgcagc ccgggctggg 721 cataaaagtc agggcagagc catctattgc ttacatttgc ttctagcctg caggtcgagg 781 agcgcagcct tccagaagca gagcgcggcg ccttaagctg cagaagttgg tcgtgaggca 841 ctgggcaggt aagtatcaag gttacaagac aggtttaagg agaccaatag aaactgggct 901 tgtcgagaca gagaagactc ttgcgtttct gataggcacc tattggtctt actgacatcc 961 actttgcctt tctctccaca ggtgtccact cccagttcaa ttacagctct taagaaacta 1021 gtagccacca tgtctagtga tgatgaggct actgctgact ctcaacattc tactcctcca 1081 aaaaagaaga gaaaggtaga agaccccatg gtgtctaagg gcgaagagct gatcaaggaa 1141 aatatgcgta tgaaggtggt catggaaggt tcggtcaacg gccaccaatt caaatgcaca 1201 ggtgaaggag aaggcaatcc gtacatggga actcaaacca tgaggatcaa agtcatcgag 1261 ggaggacccc tgccatttgc ctttgacatt cttgccacgt cgttcatgta tggcagccgt 1321 acttttatca agtacccgaa aggcattcct gatttcttta aacagtcctt tcctgagggt 1381 tttacttggg aaagagttac gagatacgaa gatggtggag tcgtcaccgt catgcaggac 1441 accagccttg aggatggctg tctcgtttac cacgtccaag tcagaggggt aaactttccc 1501 tccaatggtc ccgtgatgca gaagaagacc aagggttggg agcctaatac agagatgatg 1561 tatccagcag atggtggtct gaggggatac actcatatgg cactgaaagt tgatggtggt 1621 ggccatctgt cttgctcttt cgtaacaact tacaggtcaa aaaagaccgt cgggaacatc 1681 aagatgcccg gtatccatgc cgttgatcac cgcctggaaa ggttagagga aagtgacaat 1741 gaaatgttcg tagtacaacg cgaacacgca gttgccaagt tcgccgggct tggtggtggg 1801 atggacgagc tgtacaagta aggcgcgcca cccctgcagg gaattcgata tcaagcttat 1861 cgataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt 1921 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc 1981 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga 2041 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc 2101 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct 2161 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg 2221 gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct 2281 gctcgcctat gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc 2341 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg 2401 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcatcgata 2461 ccgagcgctg ctcgagagat ctgtgatagc ggccatcaag ctggccgcga ctctagatca 2521 taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc 2581 ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt 2641 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac 2701 tgcattctag ttgtggtttg tccaaactca tcaatgtatc agcttatcga taccgcatgc 2761 acgtgcggac cgagcggccg caggaacccc tagtgatgga gttggccact ccctctctgc 2821 gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc 2881 gggcggcctc agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt 2941 ttctccttac gcatctgtgc ggtatttcac accgcatacg tcaaagcaac catagtacgc 3001 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac 3061 acttgccagc gccttagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt 3121 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc 3181 tttacggcac ctcgacccca aaaaacttga tttgggtgat ggttcacgta gtgggccatc 3241 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact 3301 cttgttccaa actggaacaa cactcaactc tatctcgggc tattcttttg atttataagg 3361 gattttgccg atttcggtct attggttaaa aaatgagctg atttaacaaa aatttaacgc 3421 gaattttaac aaaatattaa cgtttacaat tttatggtgc actctcagta caatctgctc 3481 tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg 3541 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat 3601 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg 3661 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt 3721 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta 3781 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat 3841 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt 3901 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg 3961 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga 4021 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg 4081 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt 4141 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg 4201 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg 4261 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga 4321 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc 4381 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc 4441 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc 4501 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg 4561 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac 4621 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc 4681 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt 4741 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac 4801 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa 4861 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc 4921 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt 4981 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg 5041 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc 5101 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt 5161 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga 5221 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct 5281 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg 5341 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca 5401 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa 5461 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgt //