LOCUS Exported 5259 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION An AAV genome that expresses the fluorescent protein mTurquoise2 from the hSyn1 promoter. ACCESSION . VERSION . KEYWORDS pAAV-hSyn1-mTurquoise2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5259) 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 5259) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5259 /organism="synthetic DNA construct" /mol_type="other DNA" repeat_region 1..130 /label=AAV2 ITR (alternate) /note="Functional equivalent of wild-type AAV2 ITR" promoter 175..622 /label=hSyn promoter /note="human synapsin I promoter; confers neuron-specific expression (Kugler et al., 2003)" regulatory 636..645 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 642..1361 /codon_start=1 /product="enhanced monomeric variant of CFP (Goedhart et al., 2012)" /label=mTurquoise2 /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 642..1361 /codon_start=1 /product="enhanced monomeric variant of CFP (Goedhart et al., 2010)" /label=mTurquoise /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(687..708) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(948..967) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1295..1316 /label=EGFP-C /note="EGFP, forward primer" misc_feature 1386..1974 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(1439..1459) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(1857..1868) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 2006..2482 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(2205..2224) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" repeat_region 2522..2662 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 2522..2651 /label=AAV2 ITR rep_origin 2737..3192 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2824..2843) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3034..3055 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3209..3228) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 3328..3350 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(3388..3406) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 3474..3578 /gene="bla" /label=AmpR promoter CDS 3579..4439 /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(3797..3816) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 4610..5198 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5099..5118 /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 gcgtgtgtct agactgcaga gggccctgcg tatgagtgca 181 agtgggtttt aggaccagga tgaggcgggg tgggggtgcc tacctgacga ccgaccccga 241 cccactggac aagcacccaa cccccattcc ccaaattgcg catcccctat cagagagggg 301 gaggggaaac aggatgcggc gaggcgcgtg cgcactgcca gcttcagcac cgcggacagt 361 gccttcgccc ccgcctggcg gcgcgcgcca ccgccgcctc agcactgaag gcgcgctgac 421 gtcactcgcc ggtcccccgc aaactcccct tcccggccac cttggtcgcg tccgcgccgc 481 cgccggccca gccggaccgc accacgcgag gcgcgagata ggggggcacg ggcgcgacca 541 tctgcgctgc ggcgccggcg actcagcgct gcctcagtct gcggtgggca gcggaggagt 601 cgtgtcgtgc ctgagagcgc agtcgagaag gtaccgccac catggtgagc aagggcgagg 661 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca 721 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt 781 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgtcct 841 ggggcgtgca gtgcttcgcc cgctaccccg accacatgaa gcagcacgac ttcttcaagt 901 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact 961 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga 1021 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactact 1081 tcagcgacaa cgtctatatc accgccgaca agcagaagaa cggcatcaag gccaacttca 1141 agatccgcca caacatcgag gacggcggcg tgcagctcgc cgaccactac cagcagaaca 1201 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtcca 1261 agctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg 1321 ccgccgggat cactctcggc atggacgagc tgtacaagtg agaattcgat atcaagctta 1381 tcgataatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg 1441 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt 1501 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg 1561 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc 1621 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc 1681 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc 1741 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc 1801 tgctcgccta tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg 1861 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc 1921 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcatcgat 1981 accgagcgct gctcgagaga tctacgggtg gcatccctgt gacccctccc cagtgcctct 2041 cctggccctg gaagttgcca ctccagtgcc caccagcctt gtcctaataa aattaagttg 2101 catcattttg tctgactagg tgtccttcta taatattatg gggtggaggg gggtggtatg 2161 gagcaagggg caagttggga agacaacctg tagggcctgc ggggtctatt gggaaccaag 2221 ctggagtgca gtggcacaat cttggctcac tgcaatctcc gcctcctggg ttcaagcgat 2281 tctcctgcct cagcctcccg agttgttggg attccaggca tgcatgacca ggctcagcta 2341 atttttgttt ttttggtaga gacggggttt caccatattg gccaggctgg tctccaactc 2401 ctaatctcag gtgatctacc caccttggcc tcccaaattg ctgggattac aggcgtgaac 2461 cactgctccc ttccctgtcc ttctgatttt gtaggtaacc acgtgcggac cgagcggccg 2521 caggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag 2581 gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc agtgagcgag 2641 cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt ttctccttac gcatctgtgc 2701 ggtatttcac accgcatacg tcaaagcaac catagtacgc gccctgtagc ggcgcattaa 2761 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccttagcgc 2821 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag 2881 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca 2941 aaaaacttga tttgggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc 3001 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa 3061 cactcaactc tatctcgggc tattcttttg atttataagg gattttgccg atttcggtct 3121 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa 3181 cgtttacaat tttatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc 3241 agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat 3301 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt 3361 catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg 3421 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa 3481 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac 3541 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg 3601 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc 3661 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg 3721 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga 3781 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc 3841 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag 3901 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga 3961 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg 4021 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga 4081 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt 4141 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact 4201 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt 4261 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg 4321 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta 4381 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac 4441 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta 4501 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt 4561 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt 4621 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt 4681 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc 4741 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg 4801 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg 4861 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt 4921 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac 4981 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg 5041 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg 5101 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat 5161 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt 5221 tacggttcct ggccttttgc tggccttttg ctcacatgt //