LOCUS Exported 6124 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Bacterial expression of mutant human alpha synuclein. ACCESSION . VERSION . KEYWORDS pET11a asyn Q79N (Q109H) (TAC-->TAT) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6124) TITLE Lashuel Lab plasmids REFERENCE 2 (bases 1 to 6124) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6124 /organism="synthetic DNA construct" /mol_type="other DNA" rep_origin 2..590 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 491..510 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 744..761 /label=L4440 /note="L4440 vector, forward primer" misc_feature 776..915 /label=bom /note="basis of mobility region from pBR322" primer_bind complement(841..860) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 1001..1023 /label=pGEX 3' /note="pGEX vectors, reverse primer" CDS complement(1017..1208) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(2520..3602) /codon_start=1 /gene="lacI" /product="lac repressor" /label=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)." /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR ALADSLMQLARQVSRLESGQ" primer_bind 3564..3583 /label=LacI-R /note="LacI, reverse primer" promoter complement(3603..3680) /gene="lacI" /label=lacI promoter primer_bind 3886..3905 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" primer_bind 3989..4008 /label=T7 /note="T7 promoter, forward primer" promoter 3989..4007 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 4008..4032 /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)." RBS 4047..4069 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" CDS 4503..4535 /codon_start=1 /product="leader peptide from bacteriophage T7 gene 10" /label=T7 tag (gene 10 leader) /note="promotes efficient translation in E. coli" /translation="MASMTGGQQMG" primer_bind complement(4613..4631) /label=T7 Term /note="T7 terminator, reverse primer" terminator 4627..4674 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" promoter complement(4849..4877) /label=tet promoter /note="E. coli promoter for tetracycline efflux protein gene" primer_bind complement(4904..4922) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 4990..5094 /gene="bla" /label=AmpR promoter CDS 5095..5955 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(5313..5332) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" ORIGIN 1 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac 61 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct 121 tcagcagagc gcagatacca aatactgtyc ttctagtgta gccgtagtta ggccaccact 181 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg 241 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata 301 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga 361 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag 421 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg 481 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac 541 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca 601 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg 661 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc 721 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga 781 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcaatggt gcactctcag 841 tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac 901 tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt 961 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag 1021 aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg 1081 tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc 1141 agaagcgtta atgtctggct tctgataaag cgggccatgt taagggcggt tttttcctgt 1201 ttggtcactg atgcctccgt gtaaggggga tttctgttca tgggggtaat gataccgatg 1261 aaacgagaga ggatgctcac gatacgggtt actgatgatg aacatgcccg gttactggaa 1321 cgttgtgagg gtaaacaact ggcggtatgg atgcggcggg accagagaaa aatcactcag 1381 ggtcaatgcc agcgcttcgt taatacagat gtaggtgttc cacagggtag ccagcagcat 1441 cctgcgatgc agatccggaa cataatggtg cagggcgctg acttccgcgt ttccagactt 1501 tacgaaacac ggaaaccgaa gaccattcat gttgttgctc aggtcgcaga cgttttgcag 1561 cagcagtcgc ttcacgttcg ctcgcgtatc ggtgattcat tctgctaacc agtaaggcaa 1621 ccccgccagc ctagccgggt cctcaacgac aggagcacga tcatgcgcac ccgtggccag 1681 gacccaacgc tgcccgagat gcgccgcgtg cggctgctgg agatggcgga cgcgatggat 1741 atgttctgcc aagggttggt ttgcgcattc acagttctcc gcaagaattg attggctcca 1801 attcttggag tggtgaatcc gttagcgagg tgccgccggc ttccattcag gtcgaggtgg 1861 cccggctcca tgcaccgcga cgcaacgcgg ggaggcagac aaggtatagg gcggcgccta 1921 caatccatgc caacccgttc catgtgctcg ccgaggcggc ataaatcgcc gtgacgatca 1981 gcggtccaat gatcgaagtt aggctggtaa gagccgcgag cgatccttga agctgtccct 2041 gatggtcgtc atctacctgc ctggacagca tggcctgcaa cgcgggcatc ccgatgccgc 2101 cggaagcgag aagaatcata atggggaagg ccatccagcc tcgcgtcgcg aacgccagca 2161 agacgtagcc cagcgcgtcg gccgccatgc cggcgataat ggcctgcttc tcgccgaaac 2221 gtttggtggc gggaccagtg acgaaggctt gagcgagggc gtgcaagatt ccgaataccg 2281 caagcgacag gccgatcatc gtcgcgctcc agcgaaagcg gtcctcgccg aaaatgaccc 2341 agagcgctgc cggcacctgt cctacgagtt gcatgataaa gaagacagtc ataagtgcgg 2401 cgacgatagt catgccccgc gcccaccgga aggagctgac tgggttgaag gctctcaagg 2461 gcatcggtcg agatcccggt gcctaatgag tgagctaact tacattaatt gcgttgcgct 2521 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac 2581 gcgcggggag aggcggtttg cgtattgggc gccagggtgg tttttctttt caccagtgag 2641 acgggcaaca gctgattgcc cttcaccgcc tggccctgag agagttgcag caagcggtcc 2701 acgctggttt gccccagcag gcgaaaatcc tgtttgatgg tggttaacgg cgggatataa 2761 catgagctgt cttcggtatc gtcgtatccc actaccgaga tatccgcacc aacgcgcagc 2821 ccggactcgg taatggcgcg cattgcgccc agcgccatct gatcgttggc aaccagcatc 2881 gcagtgggaa cgatgccctc attcagcatt tgcatggttt gttgaaaacc ggacatggca 2941 ctccagtcgc cttcccgttc cgctatcggc tgaatttgat tgcgagtgag atatttatgc 3001 cagccagcca gacgcagacg cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt 3061 tgctggtgac ccaatgcgac cagatgctcc acgcccagtc gcgtaccgtc ttcatgggag 3121 aaaataatac tgttgatggg tgtctggtca gagacatcaa gaaataacgc cggaacatta 3181 gtgcaggcag cttccacagc aatggcatcc tggtcatcca gcggatagtt aatgatcagc 3241 ccactgacgc gttgcgcgag aagattgtgc accgccgctt tacaggcttc gacgccgctt 3301 cgttctacca tcgacaccac cacgctggca cccagttgat cggcgcgaga tttaatcgcc 3361 gcgacaattt gcgacggcgc gtgcagggcc agactggagg tggcaacgcc aatcagcaac 3421 gactgtttgc ccgccagttg ttgtgccacg cggttgggaa tgtaattcag ctccgccatc 3481 gccgcttcca ctttttcccg cgttttcgca gaaacgtggc tggcctggtt caccacgcgg 3541 gaaacggtct gataagagac accggcatac tctgcgacat cgtataacgt tactggtttc 3601 acattcacca ccctgaattg actctcttcc gggcgctatc atgccatacc gcgaaaggtt 3661 ttgcgccatt cgatggtgtc cgggatctcg acgctctccc ttatgcgact cctgcattag 3721 gaagcagccc agtagtaggt tgaggccgtt gagcaccgcc gccgcaagga atggtgcatg 3781 caaggagatg gcgcccaaca gtcccccggc cacggggcct gccaccatac ccacgccgaa 3841 acaagcgctc atgagcccga agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat 3901 ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta 3961 gaggatcgag atctcgatcc cgcgaaatta atacgactca ctatagggga attgtgagcg 4021 gataacaatt cccctctaga aataattttg tttaacttta agaaggagat atacatatgg 4081 atgtattcat gaaaggactt tcaaaggcca aggagggagt tgtggctgct gctgagaaaa 4141 ccaaacaggg tgtggcagaa gcagcaggaa agacaaaaga gggtgttctc tatgtaggct 4201 ccaaaaccaa ggagggagtg gtgcatggtg tggcaacagt ggctgagaag accaaagagc 4261 aagtgacaaa tgttggagga gcagtggtga cgggtgtgac agcagtagcc aacaagacag 4321 tggagggagc ggggagcatt gcagcagcca ctggctttgt caaaaaggac cagttgggca 4381 agaatgaaga aggagcccca cacgaaggaa ttctggaaga tatgcctgtg gatcctgaca 4441 atgaggctta tgaaatgcct tctgaggaag ggtatcaaga ctatgaacct gaagcctaac 4501 atatggctag catgactggt ggacagcaaa tgggtcgcgg atcccgggta cctcgagtcg 4561 accgatccgg ctgctaacaa agcccgaaag gaagctgagt tggctgctgc caccgctgag 4621 caataactag cataacccct tggggcctct aaacgggtct tgaggggttt tttgctgaaa 4681 ggaggaacta tatccggata tcccgcaaga ggcccggcag taccggcata accaagccta 4741 tgcctacagc atccagggtg acggtgccga ggatgacgat gagcgcattg ttagatttca 4801 tacacggtgc ctgactgcgt tagcaattta actgtgataa actaccgcat taaagcttat 4861 cgatgataag ctgtcaaaca tgagaattct tgaagacgaa agggcctcgt gatacgccta 4921 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg 4981 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg 5041 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt 5101 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt 5161 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg 5221 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa 5281 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt 5341 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag 5401 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt 5461 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga 5521 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt 5581 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgca 5641 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg 5701 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc 5761 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt 5821 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg 5881 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg 5941 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa 6001 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa 6061 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga 6121 tctt //