LOCUS Exported 5606 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION AAV-mediated expression of mCherry under the GFAP promoter. ACCESSION . VERSION . KEYWORDS pAAV-GFAP104-mCherry SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5606) AUTHORS Perea G, Yang A, Boyden ES, Sur M TITLE Optogenetic astrocyte activation modulates response selectivity of visual cortex neurons in vivo. JOURNAL Nat Commun. 2014;5:3262. doi: 10.1038/ncomms4262. PUBMED 24500276 REFERENCE 2 (bases 1 to 5606) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5606 /organism="synthetic DNA construct" /mol_type="other DNA" repeat_region 1..130 /label=AAV2 ITR (alternate) /note="Functional equivalent of wild-type AAV2 ITR" regulatory 998..1007 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1004..1714 /codon_start=1 /product="monomeric derivative of DsRed fluorescent protein (Shaner et al., 2004)" /label=mCherry /note="mammalian codon-optimized" /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA EGRHSTGGMDELYK" primer_bind complement(1149..1167) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 1417..1436 /label=mCherry-F /note="mCherry, forward primer" misc_feature 1739..2327 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(1792..1812) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(2210..2221) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 2347..2823 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(2546..2565) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" repeat_region 2869..3009 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 2869..2998 /label=AAV2 ITR rep_origin 3084..3539 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3171..3190) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3381..3402 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3556..3575) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 3675..3697 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(3735..3753) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 3821..3925 /gene="bla" /label=AmpR promoter CDS 3926..4786 /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(4144..4163) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 4957..5545 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5446..5465 /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 gcgttgcaaa gatggataaa gttttaaaca gagaggaatc 181 tttgcagcta atggaccttc taggtcttga aaggagtggg aattggctcc ggtgcccgtc 241 agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg gtcggcagcg 301 ctagatctaa catatcctgg tgtggagtag gggacgctgc tctgacagag gctcgggggc 361 ctgagctggc tctgtgagct ggggaggagg cagacagcca ggccttgtct gcaagcagac 421 ctggcagcat tgggctggcc gccccccagg gcctcctctt catgcccagt gaatgactca 481 ccttggcaca gacacaatgt tcggggtggg cacagtgcct gcttcccgcc gcaccccagc 541 ccccctcaaa tgccttccga gaagcccatt gagcaggggg cttgcattgc accccagcct 601 gacagcctgg catcttggga taaaagcagc acagccccct aggggctgcc cttgctgtgt 661 ggcgccaccg gcggtggaga acaaggctct attcagcctg tgcccaggaa aggggatcag 721 gggatgccca ggcatggaca gtgggtggca gggggggaga ggagggctgt ctgcttccca 781 gaagtccaag gacacaaatg ggtgagggga gagctctccc catagctggg ctgcggccca 841 accccacccc ctcaggctat gccagggggt gttgccaggg gcacccgggc atcgccagtc 901 tagcccactc cttcataaag ccctcgcatc ccaggagcga gcagagccag agcaggttgg 961 agaggagacg catcacctcc gctgctcgcg gggatccgcc accatggtga gcaagggcga 1021 ggaggataac atggccatca tcaaggagtt catgcgcttc aaggtgcaca tggagggctc 1081 cgtgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac 1141 ccagaccgcc aagctgaagg tgaccaaggg tggccccctg cccttcgcct gggacatcct 1201 gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag caccccgccg acatccccga 1261 ctacttgaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga 1321 cggcggcgtg gtgaccgtga cccaggactc ctccctgcag gacggcgagt tcatctacaa 1381 ggtgaagctg cgcggcacca acttcccctc cgacggcccc gtaatgcaga agaagaccat 1441 gggctgggag gcctcctccg agcggatgta ccccgaggac ggcgccctga agggcgagat 1501 caagcagagg ctgaagctga aggacggcgg ccactacgac gctgaggtca agaccaccta 1561 caaggccaag aagcccgtgc agctgcccgg cgcctacaac gtcaacatca agttggacat 1621 cacctcccac aacgaggact acaccatcgt ggaacagtac gaacgcgccg agggccgcca 1681 ctccaccggc ggcatggacg agctgtacaa gtaagaattc tgtacaaagc ttatcgataa 1741 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc 1801 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat 1861 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg 1921 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg 1981 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat 2041 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt 2101 gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc 2161 ctatgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa 2221 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg 2281 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcatc gataccggct 2341 cgagacgggt ggcatccctg tgacccctcc ccagtgcctc tcctggccct ggaagttgcc 2401 actccagtgc ccaccagcct tgtcctaata aaattaagtt gcatcatttt gtctgactag 2461 gtgtccttct ataatattat ggggtggagg ggggtggtat ggagcaaggg gcaagttggg 2521 aagacaacct gtagggcctg cggggtctat tgggaaccaa gctggagtgc agtggcacaa 2581 tcttggctca ctgcaatctc cgcctcctgg gttcaagcga ttctcctgcc tcagcctccc 2641 gagttgttgg gattccaggc atgcatgacc aggctcagct aatttttgtt tttttggtag 2701 agacggggtt tcaccatatt ggccaggctg gtctccaact cctaatctca ggtgatctac 2761 ccaccttggc ctcccaaatt gctgggatta caggcgtgaa ccactgctcc cttccctgtc 2821 cttctcgagc tgattttgta ggtaaccacg tgcggaccga gcggccgcag gaacccctag 2881 tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa 2941 aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct 3001 gcctgcaggg gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc 3061 gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt 3121 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc 3181 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg 3241 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt 3301 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt 3361 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat 3421 ctcgggctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa 3481 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt 3541 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc 3601 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca 3661 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg 3721 cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat 3781 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt 3841 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct 3901 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc 3961 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa 4021 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg 4081 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt 4141 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg 4201 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac 4261 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc 4321 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa 4381 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc 4441 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt 4501 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga 4561 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa 4621 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa 4681 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa 4741 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt 4801 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt 4861 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg 4921 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt 4981 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca 5041 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac 5101 tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac 5161 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct 5221 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg 5281 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca 5341 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt 5401 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta 5461 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc 5521 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc 5581 cttttgctgg ccttttgctc acatgt //