LOCUS Exported 5861 bp ds-DNA circular SYN 05-JAN-2023 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pHM6-alphasynuclein-A53T SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5861) AUTHORS Furlong RA, Narain Y, Rankin J, Wyttenbach A, Rubinsztein DC TITLE Alpha-synuclein overexpression promotes aggregation of mutant huntingtin. JOURNAL Biochem J. 2000 Mar 15;346 Pt 3:577-81. PUBMED 10698681 REFERENCE 2 (bases 1 to 5861) AUTHORS . TITLE Direct Submission JOURNAL Exported Jan 5, 2023 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5861 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(67..86) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 258..637 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 638..841 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 792..812 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 886..905 /label=T7 /note="T7 promoter, forward primer" promoter 886..904 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 927..953 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" CDS 1407..1424 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" promoter complement(1437..1455) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(1438..1455) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(1475..1492) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 1481..1705 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 1751..2179 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(1838..1857) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2048..2069 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(2188..2208) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 2193..2522 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 2373..2508 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2435..2454 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 2589..3383 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(2643..2662) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3253..3272 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 3557..3678 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3594..3613) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3648..3667 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(3727..3743) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3727..3743) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(3740..3762) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 3751..3767 /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)." promoter complement(3775..3805) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 3820..3841 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(3958..3975) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4129..4717) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4209..4228) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4888..5748) /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 5511..5530 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5749..5853) /gene="bla" /label=AmpR promoter ORIGIN 1 cccgaaaagt gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtgcac 61 tctcagtaca atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt 121 gttggaggtc gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac 181 cgacaattgc atgaagaatc tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg 241 ggccagatat acgcgttgac attgattatt gactagttat taatagtaat caattacggg 301 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc 361 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat 421 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc 481 ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga 541 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg 601 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat 661 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt 721 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc 781 cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc 841 tctctggcta actagagaac ccactgctta ctggcttatc gaaattaata cgactcacta 901 tagggagacc caagctggag accatgtacc catacgacgt cccagactac gctggaagct 961 tgatggatgt attcatgaaa ggactttcaa aggccaagga gggagttgtg gctgctgctg 1021 agaaaaccaa acagggtgtg gcagaagcag caggaaagac aaaagagggt gttctctatg 1081 taggctccaa aaccaaggag ggagtggtgc atggtgtgac aacagtggct gagaagacca 1141 aagagcaagt gacaaatgtt ggaggagcag tggtgacggg tgtgacagca gtagcccaga 1201 agacagtgga gggagcaggg agcattgcag cagccactgg ctttgtcaaa aaggaccagt 1261 tgggcaagaa tgaagaagga gccccacagg aaggaattct ggaagatatg cctgtggatc 1321 ctgacaatga ggcttatgaa atgccttctg aggaagggta tcaagactac gaacctgaag 1381 ccgagaattc gcggccgcta cacgtgcatc atcatcatca tcattaaggg ccctattcta 1441 tagtgtcacc taaatgctag agctcgctga tcagcctcga ctgtgccttc tagttgccag 1501 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact 1561 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt 1621 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat 1681 gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg 1741 gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc 1801 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc 1861 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg 1921 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca 1981 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc 2041 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct 2101 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa 2161 caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc 2221 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt 2281 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt 2341 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg 2401 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct 2461 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca 2521 aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat 2581 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga 2641 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc 2701 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga 2761 atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg 2821 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc 2881 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg 2941 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga 3001 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc 3061 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca 3121 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg 3181 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct 3241 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg 3301 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc 3361 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac 3421 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt 3481 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga 3541 gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag 3601 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa 3661 actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta 3721 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat 3781 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt 3841 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta 3901 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc 3961 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa 4021 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa 4081 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct 4141 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac 4201 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc 4261 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc 4321 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg 4381 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga 4441 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag 4501 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta 4561 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag 4621 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg 4681 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac 4741 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc 4801 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag 4861 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc 4921 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac 4981 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc 5041 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg 5101 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag 5161 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc 5221 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac 5281 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag 5341 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac 5401 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg 5461 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc 5521 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact 5581 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg 5641 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa 5701 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt 5761 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg 5821 tatttagaaa aataaacaaa taggggttcc gcgcacattt c //