LOCUS Exported 6891 bp ds-DNA circular SYN 17-JUN-2022 DEFINITION AAV vector expressing GFP. ACCESSION . VERSION . KEYWORDS AAV-CMV-GFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6891) AUTHORS Xiong W, MacColl Garfinkel AE, Li Y, Benowitz LI, Cepko CL TITLE NRF2 promotes neuronal survival in neurodegeneration and acute nerve damage. JOURNAL J Clin Invest. 2015 Apr;125(4):1433-45. doi: 10.1172/JCI79735. Epub 2015 Mar 23. PUBMED 25798616 REFERENCE 2 (bases 1 to 6891) AUTHORS . TITLE Direct Submission JOURNAL Exported Jun 17, 2022 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6891 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 10..389 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 390..593 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 544..564 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 590..614 /label=LNCX /note="Human CMV promoter, forward primer" intron 707..1182 /label=-beta-globin intron /note="internally truncated intron from human beta-globin" regulatory 1260..1269 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1266..1985 /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 1266..1982 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(1311..1332) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(1572..1591) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1919..1940 /label=EGFP-C /note="EGFP, forward primer" primer_bind complement(2015..2031) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2016..2032) /label=pBluescriptKS /note="For pBluescript vector" polyA_signal 2125..2519 /label=-beta-globin poly(A) /note="human beta-globin polyadenylation signal" repeat_region 2874..3003 /label=AAV2 ITR rep_origin 3374..3887 /direction=RIGHT /label=M13 ori /note="M13 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3461..3480) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3671..3692 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4369..4388) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 4488..4510 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(4548..4566) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 4634..4738 /gene="bla" /label=AmpR promoter CDS 4739..5599 /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(4957..4976) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 5770..6358 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6259..6278 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 6512..6529 /label=L4440 /note="L4440 vector, forward primer" repeat_region 6595..6724 /label=AAV2 ITR primer_bind complement(6780..6799) /label=SV40pro-F /note="SV40 promoter/origin, forward primer" ORIGIN 1 cgccatgttg acattgatta ttgactagtt attaatagta atcaattacg gggtcattag 61 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct 121 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc 181 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg 241 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat 301 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca 361 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc 421 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga 481 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat 541 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctcgtttag 601 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagacacc 661 gggaccgatc cagcctcccc tcgaagctga tcctgagaac ttcagggtga gtctatggga 721 cccttgatgt tttctttccc cttcttttct atggttaagt tcatgtcata ggaaggggag 781 aagtaacagg gtacacatat tgaccaaatc agggtaattt tgcatttgta attttaaaaa 841 atgctttctt cttttaatat acttttttgt ttatcttatt tctaatactt tccctaatct 901 ctttctttca gggcaataat gatacaatgt atcatgcctc tttgcaccat tctaaagaat 961 aacagtgata atttctgggt taaggcaata gcaatatttc tgcatataaa tatttctgca 1021 tataaattgt aactgatgta agaggtttca tattgctaat agcagctaca atccagctac 1081 cattctgctt ttattttatg gttgggataa ggctggatta ttctgagtcc aagctaggcc 1141 cttttgctaa tcatgttcat acctcttatc ttcctcccac agctcctggg caacgtgctg 1201 gtctgtgtgc tggcccatca ctttggcaaa gaattccgcg ggcccgggat ccaccggtcg 1261 ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc 1321 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca 1381 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc 1441 ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca 1501 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca 1561 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca 1621 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg 1681 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga 1741 agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc 1801 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca 1861 accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca 1921 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca 1981 agtaaagcgg ccgctctaga ggatccaagc ttatcgatac cgtcgacctc gagggcccag 2041 atctaattca ccccaccagt gcaggctgcc tatcagaaag tggtggctgg tgtggctaat 2101 gccctggccc acaagtatca ctaagctcgc tttcttgctg tccaatttct attaaaggtt 2161 cctttgttcc ctaagtccaa ctactaaact gggggatatt atgaagggcc ttgagcatct 2221 ggattctgcc taataaaaaa catttatttt cattgcaatg atgtatttaa attatttctg 2281 aatattttac taaaaaggga atgtgggagg tcagtgcatt taaaacataa agaaatgaag 2341 agctagttca aaccttggga aaatacacta tatcttaaac tccatgaaag aaggtgaggc 2401 tgcaaacagc taatgcacat tggcaacagc ccctgatgcc tatgccttat tcatccctca 2461 gaaaaggatt caagtagagg cttgatttgg aggttaaagt tttgctatgc tgtattttac 2521 attacttatt gttttagctg tcctcatgaa tgtcttttca ctacccattt gcttatcctg 2581 catctctcag ccttgactcc actcagttct cttgcttaga gataccacct ttcccctgaa 2641 gtgttccttc catgttttac ggcgagatgg tttctcctcg cctggccact cagccttagt 2701 tgtctctgtt gtcttataga ggtctacttg aagaaggaaa aacagggggc atggtttgac 2761 tgtcctgtga gcccttcttc cctgcctccc ccactcacag tgacccggaa tccctcgaca 2821 tctagagcat ggctacgtag ataagtagca tggcgggtta atcattaact acaaggaacc 2881 cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg 2941 accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg 3001 cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct 3061 gaatggcgaa tggcgattcc gttgcaatgg ctggcggtaa tattgttctg gatattacca 3121 gcaaggccga tagtttgagt tcttctactc aggcaagtga tgttattact aatcaaagaa 3181 gtattgcgac aacggttaat ttgcgtgatg gacagactct tttactcggt ggcctcactg 3241 attataaaaa cacttctcag gattctggcg taccgttcct gtctaaaatc cctttaatcg 3301 gcctcctgtt tagctcccgc tctgattcta acgaggaaag cacgttatac gtgctcgtca 3361 aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg 3421 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct 3481 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta 3541 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt 3601 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg 3661 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat 3721 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt 3781 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattta aatatttgct 3841 tatacaatct tcctgttttt ggggcttttc tgattatcaa ccggggtaca tatgattgac 3901 atgctagttt tacgattacc gttcatcgat tctcttgttt gctccagact ctcaggcaat 3961 gacctgatag cctttgtaga gacctctcaa aaatagctac cctctccggc atgaatttat 4021 cagctagaac ggttgaatat catattgatg gtgatttgac tgtctccggc ctttctcacc 4081 cgtttgaatc tttacctaca cattactcag gcattgcatt taaaatatat gagggttcta 4141 aaaattttta tccttgcgtt gaaataaagg cttctcccgc aaaagtatta cagggtcata 4201 atgtttttgg tacaaccgat ttagctttat gctctgaggc tttattgctt aattttgcta 4261 attctttgcc ttgcctgtat gatttattgg atgttggaat cgcctgatgc ggtattttct 4321 ccttacgcat ctgtgcggta tttcacaccg catatggtgc actctcagta caatctgctc 4381 tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg 4441 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat 4501 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg 4561 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt 4621 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta 4681 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat 4741 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt 4801 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg 4861 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga 4921 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg 4981 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt 5041 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg 5101 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg 5161 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga 5221 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc 5281 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc 5341 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc 5401 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg 5461 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac 5521 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc 5581 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt 5641 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac 5701 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa 5761 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc 5821 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt 5881 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg 5941 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc 6001 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt 6061 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga 6121 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct 6181 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg 6241 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca 6301 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa 6361 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt 6421 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga 6481 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga 6541 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctgcgc 6601 gctcgctcgc tcactgaggc cgcccgggca aagcccgggc gtcgggcgac ctttggtcgc 6661 ccggcctcag tgagcgagcg agcgcgcaga gagggagtgg ccaactccat cactaggggt 6721 tccttgtagt taatgattaa cccgccatgc tacttatcta cgtagccatg ctctagcggc 6781 ctcggcctct gcataaataa aaaaaattag tcagccatga gcttggccca ttgcatacgt 6841 tgtatccata tcataatatg tacatttata ttggctcatg tccaacatta c //