LOCUS Exported 5925 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION expression of M87 N184X spastin in mammalian cells. ACCESSION . VERSION . KEYWORDS pCMV-Tag/M87 N184X SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5925) AUTHORS Solowska JM, Rao AN, Baas PW TITLE Studies on truncating mutations of SPAST associated with Hereditary Spastic Paraplegia indicate greater accumulation and toxicity of the M1 isoform of spastin. JOURNAL Mol Biol Cell. 2017 May 11. pii: mbc.E17-01-0047. doi: 10.1091/mbc.E17-01-0047. PUBMED 28495799 REFERENCE 2 (bases 1 to 5925) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5925 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 1..304 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 305..508 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 459..479 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" promoter 554..572 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" regulatory 599..608 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" primer_bind complement(2288..2304) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2289..2305) /label=pBluescriptKS /note="For pBluescript vector" primer_bind complement(2360..2379) /label=T7 /note="T7 promoter, forward primer" promoter complement(2361..2379) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" polyA_signal 2653..2774 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2690..2709) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2744..2763 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(2781..3236) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2918..2939) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 3130..3149 /label=F1ori-R /note="F1 origin, reverse primer" promoter 3263..3367 /gene="bla" /label=AmpR promoter promoter 3369..3726 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(3392..3412) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 3577..3712 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3639..3658 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3761..4555 /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 GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(3815..3834) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4425..4444 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(4743..4762) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 4787..4834 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 5163..5751 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5652..5671 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt 61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca 121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc 181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta 241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac 301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg 361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg 421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt 481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgatt 541 acgccaagct cgaaattaac cctcactaaa gggaacaaaa gctggaggct tctcccgcgc 601 caccatggca gccaagagga gctccggggc cgcgccagca cctgcctcgg cctcggcccc 661 ggcgccggtg ccgggcggcg aggccgagcg cgtccgagtc ttccacaaac aggccttcga 721 gtacatctcc attgccctgc gcatcgatga ggatgagaaa gcaggacaga aggagcaagc 781 tgtggaatgg tataagaaag gtattgaaga actggaaaaa ggaatagctg ttatagttac 841 aggacaaggt gaacagtgtg aaagagctag acgccttcaa gctaaaatga tgacttaatt 901 tggttatggc caaggaccgc ttacaacttc tagagaagat gcaaccagtt ttgccatttt 961 ccaagtcaca aacggacgtc tataatgaca gtactaactt ggcatgccgc aatggacatc 1021 tccagtcaga aagtggagct gttccaaaaa gaaaagaccc cttaacacac actagtaatt 1081 cactgcctcg ttcaaaaaca gttatgaaaa ctggatctgc aggcctttca ggccaccata 1141 gagcacctag ttacagtggt ttatccatgg tttctggagt gaaacaggga tctggtcctg 1201 ctcctaccac tcataagggt actccgaaaa caaataggac aaataaacct tctaccccta 1261 caactgctac tcgtaagaaa aaagacttga agaattttag gaatgtggac agcaaccttg 1321 ctaaccttat aatgaatgaa attgtggaca atggaacagc tgttaaattt gatgatatag 1381 ctggtcaaga cttggcaaaa caagcattgc aagaaattgt tattcttcct tctctgaggc 1441 ctgagttgtt cacagggctt agagctcctg ccagagggct gttactcttt ggtccacctg 1501 ggaatgggaa gacaatgctg gctaaagcag tagctgcaga atcgaatgca accttcttta 1561 atataagtgc tgcaagttta acttcaaaat acgtgggaga aggagagaaa ttggtgaggg 1621 ctctttttgc tgtggctcga gaacttcaac cttctataat ttttatagat gaagttgata 1681 gccttttgtg tgaaagaaga gaaggggagc acgatgctag tagacgccta aaaactgaat 1741 ttctaataga atttgatggt gtacagtctg ctggagatga cagagtactt gtaatgggtg 1801 caactaatag gccacaagag cttgatgagg ctgttctcag gcgtttcatc aaacgggtat 1861 atgtgtcttt accaaatgag gagacaagac tacttttgct taaaaatctg ttatgtaaac 1921 aaggaagtcc attgacccaa aaagaactag cacaacttgc tagaatgact gatggatact 1981 caggaagtga cctaacagct ttggcaaaag atgcagcact gggtcctatc cgagaactaa 2041 aaccagaaca ggtgaagaat atgtctgcca gtgagatgag aaatattcga ttatctgact 2101 tcactgaatc cttgaaaaaa ataaaacgca gcgtcagccc tcaaacttta gaagcgtaca 2161 tacgttggaa caaggacttt ggagatacca ctgtttaagg aaataccttt gtaaacctgc 2221 agaacatttt acttaaaaga ggaaacacaa gatcccccgg gctgcaggaa ttcgatatca 2281 agcttatcga taccgtcgac ctcgaggggg ggcccggtac cttaattaat taaggtacca 2341 ggtaagtgta cccaattcgc cctatagtga gtcgtattac aattcactcg atcgcccttc 2401 ccaacagttg cgcagcctga atggcgaatg gagatccaat ttttaagtgt ataatgtgtt 2461 aaactactga ttctaattgt ttgtgtattt tagattcaca gtcccaaggc tcatttcagg 2521 cccctcagtc ctcacagtct gttcatgatc ataatcagcc ataccacatt tgtagaggtt 2581 ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca 2641 attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc 2701 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc 2761 atcaatgtat cttaacgcgt aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa 2821 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa 2881 atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact 2941 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc 3001 actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa 3061 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc 3121 gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt 3181 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg 3241 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc 3301 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag 3361 gaagaatcct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag 3421 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc 3481 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat 3541 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt 3601 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc 3661 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt 3721 tgcaaagatc gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt 3781 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca 3841 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct 3901 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caagacgagg cagcgcggct 3961 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc 4021 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct 4081 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga 4141 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg 4201 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gaacatcagg ggctcgcgcc 4261 agccgaactg ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac 4321 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat 4381 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga 4441 tattgctgaa gaacttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc 4501 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg 4561 actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat 4621 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg 4681 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccctag ggggaggcta 4741 actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga 4801 cagaataaaa cgcacggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc 4861 tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc 4921 ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg 4981 gcggcaggcc ctgccatagc ctcaggttac tcatatatac tttagattga tttaaaactt 5041 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc 5101 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct 5161 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta 5221 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc 5281 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac 5341 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct 5401 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat 5461 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg 5521 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa 5581 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg 5641 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga 5701 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc 5761 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct 5821 gcgttatccc ctgattctgt ggataaccgt attaccgcca tgcattagtt attaatagta 5881 atcaattacg gggtcattag ttcatagccc atatatggag ttccg //