LOCUS Exported 5683 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION AAV expression of EGFP in astrocytes. ACCESSION . VERSION . KEYWORDS pAAV.GFA104.PI.eGFP.WPRE.bGH SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5683) AUTHORS Ortinski PI, Dong J, Mungenast A, Yue C, Takano H, Watson DJ, Haydon PG, Coulter DA TITLE Selective induction of astrocytic gliosis generates deficits in neuronal inhibition. JOURNAL Nat Neurosci. 2010 May;13(5):584-91. doi: 10.1038/nn.2535. Epub 2010 Apr 25. PUBMED 20418874 REFERENCE 2 (bases 1 to 5683) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5683 /organism="synthetic DNA construct" /mol_type="other DNA" intron 816..948 /label=chimeric intron /note="chimera between introns from human beta-globin and immunoglobulin heavy chain genes" primer_bind 931..950 /label=pMT2-F /note="Synthetic intron, forward primer" regulatory 960..969 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 966..1685 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 966..1682 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(1011..1032) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(1272..1291) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1619..1640 /label=EGFP-C /note="EGFP, forward primer" primer_bind complement(1752..1772) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(2170..2181) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind complement(2248..2265) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2254..2461 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" repeat_region 2549..2678 /label=AAV2 ITR primer_bind complement(2697..2714) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(2697..2713) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2706..2728) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 2855..3310 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2942..2961) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3152..3173 /label=F1ori-F /note="F1 origin, forward primer" promoter 3336..3440 /gene="bla" /label=AmpR promoter CDS 3441..4301 /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(3659..3678) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 4472..5060 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4961..4980 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 5214..5231 /label=L4440 /note="L4440 vector, forward primer" protein_bind 5348..5369 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 5384..5414 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5422..5438 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." primer_bind 5427..5449 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 5446..5462 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 5446..5462 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" repeat_region 5500..5629 /label=AAV2 ITR ORIGIN 1 agatcggaat tcgcccttaa gctagccgag ctcggtacca gatctaacat atcctggtgt 61 ggagtagggg acgctgctct gacagaggct cgggggcctg agctggctct gtgagctggg 121 gaggaggcag acagccaggc cttgtctgca agcagacctg gcagcattgg gctggccgcc 181 ccccagggcc tcctcttcat gcccagtgaa tgactcacct tggcacagac acaatgttcg 241 gggtgggcac agtgcctgct tcccgccgca ccccagcccc cctcaaatgc cttccgagaa 301 gcccattgag cagggggctt gcattgcacc ccagcctgac agcctggcat cttgggataa 361 aagcagcaca gccccctagg ggctgccctt gctgtgtggc gccaccggcg gtggagaaca 421 aggctctatt cagcctgtgc ccaggaaagg ggatcagggg atgcccaggc atggacagtg 481 ggtggcaggg ggggagagga gggctgtctg cttcccagaa gtccaaggac acaaatgggt 541 gaggggagag ctctccccat agctgggctg cggcccaacc ccaccccctc aggctatgcc 601 agggggtgtt gccaggggca cccgggcatc gccagtctag cccactcctt cataaagccc 661 tcgcatccca ggagcgagca gagccagagc aggttggaga ggagacgcat cacctccgct 721 gctcgcgggg atcctctaga gtcgacggat ccggggaatt ccccagtttc aggatccacc 781 atggctgcag aagttggtcg tgaggcactg ggcaggtaag tatcaaggtt acaagacagg 841 tttaaggaga ccaatagaaa ctgggcttgt cgagacagag aagactcttg cgtttctgat 901 aggcacctat tggtcttact gacatccact ttgcctttct ctccacaggt gtccaggcgg 961 ccgccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc 1021 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca 1081 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc 1141 ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca 1201 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca 1261 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca 1321 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg 1381 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga 1441 agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc 1501 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca 1561 accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca 1621 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca 1681 agtaataagc ttggatccaa tcaacctctg gattacaaaa tttgtgaaag attgactggt 1741 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat 1801 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg 1861 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt 1921 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact 1981 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc 2041 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg 2101 tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc 2161 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg 2221 cggcctcttc cgcgtcttcg agatctgcct cgactgtgcc ttctagttgc cagccatctg 2281 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt 2341 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg 2401 gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg 2461 actcgagtta agggcgaatt cccgataagg atcttcctag agcatggcta cgtagataag 2521 tagcatggcg ggttaatcat taactacaag gaacccctag tgatggagtt ggccactccc 2581 tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc 2641 tttgcccggg cggcctcagt gagcgagcga gcgcgcagcc ttaattaacc taattcactg 2701 gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt 2761 gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct 2821 tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc cctgtagcgg cgcattaagc 2881 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc 2941 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct 3001 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa 3061 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc 3121 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca 3181 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat 3241 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg 3301 cttacaattt aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt 3361 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat 3421 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt 3481 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg 3541 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga 3601 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc 3661 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac 3721 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg 3781 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca 3841 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg 3901 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg 3961 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg 4021 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag 4081 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg 4141 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct 4201 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac 4261 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact 4321 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga 4381 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt 4441 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct 4501 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc 4561 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc 4621 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc 4681 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg 4741 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt 4801 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg 4861 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg 4921 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt 4981 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag 5041 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt 5101 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta 5161 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt 5221 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc 5281 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca 5341 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc 5401 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg 5461 accatgatta cgccagattt aattaaggcc ttaattaggc tgcgcgctcg ctcgctcact 5521 gaggccgccc gggcaaagcc cgggcgtcgg gcgacctttg gtcgcccggc ctcagtgagc 5581 gagcgagcgc gcagagaggg agtggccaac tccatcacta ggggttcctt gtagttaatg 5641 attaacccgc catgctactt atctacgtag ccatgctcta gga //