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" primer_bind 1..23 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(61..79) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 147..251 /gene="bla" /label=AmpR promoter CDS 252..1112 /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(470..489) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 1283..1871 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 1772..1791 /label=pBR322ori-F /note="pBR322 origin, forward primer" repeat_region 1933..2062 /label=AAV2 ITR (alternate) /note="Functional equivalent of wild-type AAV2 ITR" regulatory 2930..2939 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 2936..3646 /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(3081..3099) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 3349..3368 /label=mCherry-F /note="mCherry, forward primer" misc_feature 3671..4259 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(3724..3744) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(4142..4153) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 4279..4755 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(4478..4497) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" repeat_region 4801..4941 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 4801..4930 /label=AAV2 ITR rep_origin 5016..5471 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5103..5122) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 5313..5334 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(5488..5507) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" ORIGIN 1 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg 61 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt 121 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac 181 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa 241 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat 301 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc 361 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga 421 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg 481 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc 541 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag 601 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc 661 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg 721 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg 781 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac 841 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac 901 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg 961 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg 1021 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg 1081 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac 1141 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg 1201 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg 1261 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc 1321 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc 1381 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt 1441 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc 1501 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact 1561 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac 1621 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag 1681 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg 1741 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg 1801 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga 1861 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt 1921 ttgctcacat gtcctgcagg cagctgcgcg ctcgctcgct cactgaggcc gcccgggcgt 1981 cgggcgacct ttggtcgccc ggcctcagtg agcgagcgag cgcgcagaga gggagtggcc 2041 aactccatca ctaggggttc ctgcggccgc acgcgttgca aagatggata aagttttaaa 2101 cagagaggaa tctttgcagc taatggacct tctaggtctt gaaaggagtg ggaattggct 2161 ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag ttggggggag 2221 gggtcggcag cgctagatct aacatatcct ggtgtggagt aggggacgct gctctgacag 2281 aggctcgggg gcctgagctg gctctgtgag ctggggagga ggcagacagc caggccttgt 2341 ctgcaagcag acctggcagc attgggctgg ccgcccccca gggcctcctc ttcatgccca 2401 gtgaatgact caccttggca cagacacaat gttcggggtg ggcacagtgc ctgcttcccg 2461 ccgcacccca gcccccctca aatgccttcc gagaagccca ttgagcaggg ggcttgcatt 2521 gcaccccagc ctgacagcct ggcatcttgg gataaaagca gcacagcccc ctaggggctg 2581 cccttgctgt gtggcgccac cggcggtgga gaacaaggct ctattcagcc tgtgcccagg 2641 aaaggggatc aggggatgcc caggcatgga cagtgggtgg caggggggga gaggagggct 2701 gtctgcttcc cagaagtcca aggacacaaa tgggtgaggg gagagctctc cccatagctg 2761 ggctgcggcc caaccccacc ccctcaggct atgccagggg gtgttgccag gggcacccgg 2821 gcatcgccag tctagcccac tccttcataa agccctcgca tcccaggagc gagcagagcc 2881 agagcaggtt ggagaggaga cgcatcacct ccgctgctcg cggggatccg ccaccatggt 2941 gagcaagggc gaggaggata acatggccat catcaaggag ttcatgcgct tcaaggtgca 3001 catggagggc tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc 3061 ctacgagggc acccagaccg ccaagctgaa ggtgaccaag ggtggccccc tgcccttcgc 3121 ctgggacatc ctgtcccctc agttcatgta cggctccaag gcctacgtga agcaccccgc 3181 cgacatcccc gactacttga agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat 3241 gaacttcgag gacggcggcg tggtgaccgt gacccaggac tcctccctgc aggacggcga 3301 gttcatctac aaggtgaagc tgcgcggcac caacttcccc tccgacggcc ccgtaatgca 3361 gaagaagacc atgggctggg aggcctcctc cgagcggatg taccccgagg acggcgccct 3421 gaagggcgag atcaagcaga ggctgaagct gaaggacggc ggccactacg acgctgaggt 3481 caagaccacc tacaaggcca agaagcccgt gcagctgccc ggcgcctaca acgtcaacat 3541 caagttggac atcacctccc acaacgagga ctacaccatc gtggaacagt acgaacgcgc 3601 cgagggccgc cactccaccg gcggcatgga cgagctgtac aagtaagaat tctgtacaaa 3661 gcttatcgat aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa 3721 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat 3781 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta 3841 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc 3901 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt 3961 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg 4021 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc 4081 ttggctgctc gcctatgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc 4141 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct 4201 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgca 4261 tcgataccgg ctcgagacgg gtggcatccc tgtgacccct ccccagtgcc tctcctggcc 4321 ctggaagttg ccactccagt gcccaccagc cttgtcctaa taaaattaag ttgcatcatt 4381 ttgtctgact aggtgtcctt ctataatatt atggggtgga ggggggtggt atggagcaag 4441 gggcaagttg ggaagacaac ctgtagggcc tgcggggtct attgggaacc aagctggagt 4501 gcagtggcac aatcttggct cactgcaatc tccgcctcct gggttcaagc gattctcctg 4561 cctcagcctc ccgagttgtt gggattccag gcatgcatga ccaggctcag ctaatttttg 4621 tttttttggt agagacgggg tttcaccata ttggccaggc tggtctccaa ctcctaatct 4681 caggtgatct acccaccttg gcctcccaaa ttgctgggat tacaggcgtg aaccactgct 4741 cccttccctg tccttctcga gctgattttg taggtaacca cgtgcggacc gagcggccgc 4801 aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg 4861 ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc 4921 gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt tctccttacg catctgtgcg 4981 gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag 5041 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc 5101 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc 5161 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa 5221 aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg 5281 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac 5341 actcaactct atctcgggct attcttttga tttataaggg attttgccga tttcggtcta 5401 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac 5461 gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca 5521 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc 5581 cgcttacaga caagctgtga ccgtct //