LOCUS Exported 2900 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Bacterial expression of mutant human alpha synuclein. ACCESSION . VERSION . KEYWORDS pT7-7 asyn E46K SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 2900) AUTHORS Mbefo MK, Fares MB, Paleologou K, Oueslati A, Yin G, Tenreiro S, Pinto M, Outeiro T, Zweckstetter M, Masliah E, Lashuel HA TITLE Parkinson disease mutant E46K enhances alpha-synuclein phosphorylation in mammalian cell lines, in yeast, and in vivo. JOURNAL J Biol Chem. 2015 Apr 10;290(15):9412-27. doi: 10.1074/jbc.M114.610774. Epub 2015 Feb 5. PUBMED 25657004 REFERENCE 2 (bases 1 to 2900) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..2900 /organism="synthetic DNA construct" /mol_type="other DNA" rep_origin 9..597 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 498..517 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 751..768 /label=L4440 /note="L4440 vector, forward primer" primer_bind 1003..1025 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 1090..1109 /label=T7 /note="T7 promoter, forward primer" promoter 1090..1108 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" RBS 1140..1162 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" CDS complement(1729..2589) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 2352..2371 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(2590..2694) /gene="bla" /label=AmpR promoter primer_bind 2762..2780 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" ORIGIN 1 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca 61 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta 121 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc 181 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca 241 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta 301 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag 361 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt 421 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc 481 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac 541 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac 601 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc 661 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat 721 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag 781 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg cataggaaga 841 tcttccggaa gatcttccta tggtgcactc tcagtacaat ctgctctgat gcgctacgtg 901 actgggtcat ggctgcgccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt 961 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc 1021 agaggttttc accgtcatca ccgaaacgcg cgaggcccag cgattcgaac ttctgataga 1081 cttcgaaatt aatacgactc actataggga gaccacaacg gtttccctct agaaataatt 1141 ttgtttaact ttaagaagga gatatacata tggatgtatt catgaaagga ctttcaaagg 1201 ccaaggaggg agttgtggct gctgctgaga aaaccaaaca gggtgtggca gaagcagcag 1261 gaaagacaaa agagggtgtt ctctatgtag gctccaaaac caagaaggga gtggtgcatg 1321 gtgtggcaac agtggctgag aagaccaaag agcaagtgac aaatgttgga ggagcagtgg 1381 tgacgggtgt gacagcagta gcccagaaga cagtggaggg agcagggagc attgcagcag 1441 ccactggctt tgtcaaaaag gaccagttgg gcaagaatga agaaggagcc ccacaggaag 1501 gaattctgga agatatgcct gtggatcctg acaatgaggc ttatgaaatg ccttctgagg 1561 aagggtatca agactacgaa cctgaagcct aagaaatatc tttgctccca gtttcttgag 1621 atctgctgac agatgttcca tccaagcttg agagcttatc gatgataagc tgtcaaacat 1681 gagaattaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt 1741 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact 1801 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat 1861 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg 1921 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg 1981 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat 2041 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc 2101 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt 2161 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc 2221 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga 2281 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc 2341 gtcaacacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa 2401 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta 2461 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg 2521 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg 2581 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat 2641 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt 2701 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa 2761 aaataggcgt atcacgaggc cctttcgtct tcaagaataa aaggatctag gtgaagatcc 2821 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag 2881 accccgtaga aaagatcaaa //