LOCUS Exported 4669 bp ds-DNA circular SYN 17-JAN-2023 DEFINITION Expresses human alpha-Synuclein WT. ACCESSION . VERSION . KEYWORDS pscAAV-CBh-aSyn-WPRE3-enSV40pA SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4669) TITLE GeneDetect Michael J Fox Foundation Shared Tools Plasmids 2022 REFERENCE 2 (bases 1 to 4669) AUTHORS . TITLE Direct Submission JOURNAL Exported Jan 17, 2023 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4669 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 189..474 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer; contains an 18-bp deletion relative to the standard CMV enhancer" promoter 476..753 /label=chicken beta-actin promoter intron 754..981 /label=hybrid intron /note="hybrid between chicken beta-actin (CBA) and minute virus of mice (MMV) introns (Gray et al., 2011)" regulatory 1006..1015 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" primer_bind complement(1503..1523) /label=WPRE-R /note="WPRE, reverse primer" primer_bind complement(2024..2041) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(2024..2040) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2033..2055) /label=M13/pUC Forward /note="In lacZ gene" primer_bind complement(2249..2268) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 2368..2390 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(2428..2446) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 2514..2618 /gene="bla" /label=AmpR promoter CDS 2619..3479 /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(2837..2856) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 3650..4238 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4139..4158 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 4392..4409 /label=L4440 /note="L4440 vector, forward primer" protein_bind 4526..4547 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 4562..4592 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4600..4616 /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 4605..4627 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 4624..4640 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 4624..4640 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" ORIGIN 1 aaagcttccc ggggggatct gggccactcc ctctctgcgc gctcgctcgc tcactgaggc 61 cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg 121 agcgcgcaga gagggagtgg ccaactccat cactaggggt tcctggaggg gtggagtcgt 181 gacctaggcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc 241 cgcccattga cgtcaatagt aacgccaata gggactttcc attgacgtca atgggtggag 301 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc 361 cctattgacg tcaatgacgg taaatggccc gcctggcatt gtgcccagta catgacctta 421 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catggtcgag 481 gtgagcccca cgttctgctt cactctcccc atctcccccc cctccccacc cccaattttg 541 tatttattta ttttttaatt attttgtgca gcgatggggg cggggggggg gggggggcgc 601 gcgccagggg gggggggggg gggsgagggg gggggsgggg sgaggsggag aggtgcggcg 661 gcagccaatc agagcggcgc gctccgaaag tttcctttta tggcgaggcg gcggcggcgg 721 cggccctata aaaagcgaag cgcgcggcgg gcgggagtcg ctgcgcgctg ccttcgcccc 781 gtgccccgct ccgccgccgc ctcgcgccgc ccgccccggc tctgactgac cgcgttactc 841 ccacaggtga gcgggcggga cggcccttct cctccgggct gtaattagct gagcaagagg 901 taagggttta agggatggtt ggttggtggg gtattaatgt ttaattacct ggagcacctg 961 cctgaaatca ctttttttca ggttggaccg gctagcaccg gtgcggccgc catggatgta 1021 ttcatgaaag gactttcaaa ggccaaggag ggagttgtgg ctgctgctga gaaaaccaaa 1081 cagggtgtgg cagaagcagc aggaaagaca aaagagggtg ttctctatgt aggctccaaa 1141 accaaggagg gagtggtgca tggtgtggca acagtggctg agaagaccaa agagcaagtg 1201 acaaatgttg gaggagcagt ggtgacgggt gtgacagcag tagcccagaa gacagtggag 1261 ggagcaggga gcattgcagc agccactggc tttgtcaaaa aggaccagtt gggcaagaat 1321 gaagaaggag ccccacagga aggaattctg gaagatatgc ctgtggatcc tgacaatgag 1381 gcttatgaaa tgccttctga ggaagggtat caagactacg aacctgaagc ctaagcggcc 1441 gctaggccta atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac 1501 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt 1561 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttagt tcttgccacg 1621 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact 1681 gacaattccg tggtgtttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatc 1741 tagctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa 1801 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg 1861 ggaggttttt taaaactagt ccactccctc tctgcgcgct cgctcgctca ctgaggccgg 1921 gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc 1981 gcgcagagag ggacagatcc gggcccgcat gcgtcgacaa ttcactggcc gtcgttttac 2041 aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc 2101 ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc 2161 gcagcctgaa tggcgaatgg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta 2221 tttcacaccg catatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc 2281 agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat 2341 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt 2401 catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg 2461 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa 2521 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac 2581 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg 2641 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc 2701 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg 2761 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga 2821 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc 2881 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag 2941 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga 3001 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg 3061 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga 3121 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt 3181 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact 3241 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt 3301 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg 3361 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta 3421 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac 3481 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta 3541 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt 3601 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt 3661 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt 3721 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc 3781 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg 3841 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg 3901 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt 3961 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac 4021 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg 4081 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg 4141 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat 4201 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt 4261 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg 4321 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa 4381 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc 4441 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga 4501 aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg 4561 ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc 4621 acacaggaaa cagctatgac catgattacg ccaagctctc gagatctag //