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(44..63) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 235..614 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 615..818 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 769..789 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 863..882 /label=T7 /note="T7 promoter, forward primer" promoter 863..881 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 904..930 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" CDS 1384..1401 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" promoter complement(1414..1432) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(1415..1432) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(1452..1469) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 1458..1682 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 1728..2156 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(1815..1834) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2025..2046 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(2165..2185) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 2170..2499 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 2350..2485 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2412..2431 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 2566..3360 /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(2620..2639) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3230..3249 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 3534..3655 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3571..3590) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3625..3644 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(3704..3720) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3704..3720) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(3717..3739) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 3728..3744 /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(3752..3782) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 3797..3818 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(3935..3952) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4106..4694) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4186..4205) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4865..5725) /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 5488..5507 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5726..5830) /gene="bla" /label=AmpR promoter ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg 61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt 241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata 301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc 361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc 421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt 481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt 541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca 601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg 661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc 721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg 781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca 841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggagacc 901 atgtacccat acgacgtccc agactacgct ggaagcttga tggatgtatt catgaaagga 961 ctttcaaagg ccaaggaggg agttgtggct gctgctgaga aaaccaaaca gggtgtggca 1021 gaagcagcag gaaagacaaa agagggtgtt ctctatgtag gctccaaaac caaggaggga 1081 gtggtgcatg gtgtgacaac agtggctgag aagaccaaag agcaagtgac aaatgttgga 1141 ggagcagtgg tgacgggtgt gacagcagta gcccagaaga cagtggaggg agcagggagc 1201 attgcagcag ccactggctt tgtcaaaaag gaccagttgg gcaagaatga agaaggagcc 1261 ccacaggaag gaattctgga agatatgcct gtggatcctg acaatgaggc ttatgaaatg 1321 ccttctgagg aagggtatca agactacgaa cctgaagccg agaattcgcg gccgctacac 1381 gtgcatcatc atcatcatca ttaagggccc tattctatag tgtcacctaa atgctagagc 1441 tcgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc 1501 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga 1561 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga 1621 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat 1681 ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag 1741 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag 1801 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt 1861 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca 1921 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata 1981 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca 2041 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc 2101 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt 2161 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt 2221 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca 2281 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta 2341 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga 2401 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag 2461 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata 2521 tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat 2581 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca 2641 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg 2701 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg 2761 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact 2821 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct 2881 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg 2941 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt 3001 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc 3061 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc 3121 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga 3181 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc 3241 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt 3301 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga 3361 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt 3421 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg 3481 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt 3541 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag 3601 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg 3661 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg 3721 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta 3781 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg 3841 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga 3901 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg 3961 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag 4021 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc 4081 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca 4141 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt 4201 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc 4261 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc 4321 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc 4381 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact 4441 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg 4501 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta 4561 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca 4621 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa 4681 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg 4741 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc 4801 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg 4861 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat 4921 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg 4981 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa 5041 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca 5101 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc 5161 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt 5221 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa 5281 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat 5341 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct 5401 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga 5461 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag 5521 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga 5581 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca 5641 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg 5701 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc 5761 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag 5821 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt c //