LOCUS Exported 4533 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Inducible expression of WT M1 spastin in mammalian cells. ACCESSION . VERSION . KEYWORDS pTRE/SP M1WT SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4533) AUTHORS Solowska JM, D'Rozario M, Jean DC, Davidson MW, Marenda DR, Baas PW TITLE Pathogenic mutation of spastin has gain-of-function effects on microtubule dynamics. JOURNAL J Neurosci. 2014 Jan 29;34(5):1856-67. doi: 10.1523/JNEUROSCI.3309-13.2014. PUBMED 24478365 REFERENCE 2 (bases 1 to 4533) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4533 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 1..315 /label=tight TRE promoter /note="Tet-responsive promoter PTight, consisting of seven tet operator sequences followed by the minimal CMV promoter" protein_bind 9..27 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 45..63 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 80..98 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 116..134 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 152..170 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 187..205 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 223..241 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" primer_bind 288..312 /label=LNCX /note="Human CMV promoter, forward primer" polyA_signal 2444..2525 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2481..2500) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind complement(2536..2553) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2707..3295) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2787..2806) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3466..4326) /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 4089..4108 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4327..4431) /gene="bla" /label=AmpR promoter primer_bind 4499..4517 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" ORIGIN 1 gagtttactc cctatcagtg atagagaacg tatgtcgagt ttactcccta tcagtgatag 61 agaacgatgt cgagtttact ccctatcagt gatagagaac gtatgtcgag tttactccct 121 atcagtgata gagaacgtat gtcgagttta ctccctatca gtgatagaga acgtatgtcg 181 agtttatccc tatcagtgat agagaacgta tgtcgagttt actccctatc agtgatagag 241 aacgtatgtc gaggtaggcg tgtacggtgg gaggcctata taagcagagc tcgtttagtg 301 aaccgtcaga tcgcctggag aattcgagct cggtacccat cgcggcggcg gcagtgagag 361 gccaccatga attctccggg tggacgaggg aagaagaaag gctccggcgg cgccagcaac 421 ccggtgcctc ccaggcctcc gcccccttgc ctggcccccg cccctcccgc cgccgggccg 481 gcccctccgc ccgagtcgcc gcataagcgg aacctgtact atttctccta cccgctgttt 541 gtaggcttcg cgctgctgcg tttggtcgcc ttccacctgg ggctcctctt cgtgtggctc 601 tgccagcgct tctcccgcgc cctcatggca gccaagagga gctccggggc cgcgccagca 661 cctgcctcgg cctcggcccc ggcgccggtg ccgggcggcg aggccgagcg cgtccgagtc 721 ttccacaaac aggccttcga gtacatctcc attgccctgc gcatcgatga ggatgagaaa 781 gcaggacaga aggagcaagc tgtggaatgg tataagaaag gtattgaaga actggaaaaa 841 ggaatagctg ttatagttac aggacaaggt gaacagtgtg aaagagctag acgccttcaa 901 gctaaaatga tgactaattt ggttatggcc aaggaccgct tacaacttct agagaagatg 961 caaccagttt tgccattttc caagtcacaa acggacgtct ataatgacag tactaacttg 1021 gcatgccgca atggacatct ccagtcagaa agtggagctg ttccaaaaag aaaagacccc 1081 ttaacacaca ctagtaattc actgcctcgt tcaaaaacag ttatgaaaac tggatctgca 1141 ggcctttcag gccaccatag agcacctagt tacagtggtt tatccatggt ttctggagtg 1201 aaacagggat ctggtcctgc tcctaccact cataagggta ctccgaaaac aaataggaca 1261 aataaacctt ctacccctac aactgctact cgtaagaaaa aagacttgaa gaattttagg 1321 aatgtggaca gcaaccttgc taaccttata atgaatgaaa ttgtggacaa tggaacagct 1381 gttaaatttg atgatatagc tggtcaagac ttggcaaaac aagcattgca agaaattgtt 1441 attcttcctt ctctgaggcc tgagttgttc acagggctta gagctcctgc cagagggctg 1501 ttactctttg gtccacctgg gaatgggaag acaatgctgg ctaaagcagt agctgcagaa 1561 tcgaatgcaa ccttctttaa tataagtgct gcaagtttaa cttcaaaata cgtgggagaa 1621 ggagagaaat tggtgagggc tctttttgct gtggctcgag aacttcaacc ttctataatt 1681 tttatagatg aagttgatag ccttttgtgt gaaagaagag aaggggagca cgatgctagt 1741 agacgcctaa aaactgaatt tctaatagaa tttgatggtg tacagtctgc tggagatgac 1801 agagtacttg taatgggtgc aactaatagg ccacaagagc ttgatgaggc tgttctcagg 1861 cgtttcatca aacgggtata tgtgtcttta ccaaatgagg agacaagact acttttgctt 1921 aaaaatctgt tatgtaaaca aggaagtcca ttgacccaaa aagaactagc acaacttgct 1981 agaatgactg atggatactc aggaagtgac ctaacagctt tggcaaaaga tgcagcactg 2041 ggtcctatcc gagaactaaa accagaacag gtgaagaata tgtctgccag tgagatgaga 2101 aatattcgat tatctgactt cactgaatcc ttgaaaaaaa taaaacgcag cgtcagccct 2161 caaactttag aagcgtacat acgttggaac aaggactttg gagataccac tgtttaagga 2221 aatacctttg taaacctgca gaacatttta cttaaaagag gaaacacaag atcctctagt 2281 cagctgacgc gtgctagcgc ggccgcatcg ataagcttgt cgacgatatc tctagaggat 2341 cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct 2401 ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc 2461 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc 2521 actgcctcga gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc 2581 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg 2641 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct 2701 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca 2761 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct 2821 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc 2881 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt 2941 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc 3001 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc 3061 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg 3121 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc 3181 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag 3241 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga 3301 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat 3361 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag 3421 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat 3481 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc 3541 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat 3601 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag 3661 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg 3721 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc 3781 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca 3841 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg 3901 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc 3961 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta 4021 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc 4081 aacacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg 4141 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc 4201 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc 4261 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat 4321 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag 4381 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc 4441 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa 4501 taggcgtatc acgaggccct ttcgtcttca ctc //