LOCUS Exported 6133 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses WT myc-M1 spastin isoform in mammalian cells. ACCESSION . VERSION . KEYWORDS pCMV-Tag 3A/WT myc-M1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6133) AUTHORS Solowska JM, Garbern JY, Baas PW TITLE Evaluation of loss of function as an explanation for SPG4-based hereditary spastic paraplegia. JOURNAL Hum Mol Genet. 2010 Jul 15;19(14):2767-79. doi: 10.1093/hmg/ddq177. Epub 2010 Apr 29. PUBMED 20430936 REFERENCE 2 (bases 1 to 6133) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6133 /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 604..613 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 613..642 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" primer_bind complement(2568..2587) /label=T7 /note="T7 promoter, forward primer" promoter complement(2569..2587) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" polyA_signal 2861..2982 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2898..2917) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2952..2971 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(2989..3444) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3126..3147) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 3338..3357 /label=F1ori-R /note="F1 origin, reverse primer" promoter 3471..3575 /gene="bla" /label=AmpR promoter promoter 3577..3934 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(3600..3620) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 3785..3920 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3847..3866 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3969..4763 /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(4023..4042) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4633..4652 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(4951..4970) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 4995..5042 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 5371..5959 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5860..5879 /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 gctggagctc caccgcggtg 601 gcggccgcca tggagcagaa actcatctct gaagaggatc tggcccgggc ggatcccccg 661 ggctgcagga attctccggg tggacgaggg aagaagaaag gctccggcgg cgccagcaac 721 ccggtgcctc ccaggcctcc gcccccttgc ctggcccccg cccctcccgc cgccgggccg 781 gcccctccgc ccgagtcgcc gcataagcgg aacctgtact atttctccta cccgctgttt 841 gtaggcttcg cgctgctgcg tttggtcgcc ttccacctgg ggctcctctt cgtgtggctc 901 tgccagcgct tctcccgcgc cctcatggca gccaagagga gctccggggc cgcgccagca 961 cctgcctcgg cctcggcccc ggcgccggtg ccgggcggcg aggccgagcg cgtccgagtc 1021 ttccacaaac aggccttcga gtacatctcc attgccctgc gcatcgatga ggatgagaaa 1081 gcaggacaga aggagcaagc tgtggaatgg tataagaaag gtattgaaga actggaaaaa 1141 ggaatagctg ttatagttac aggacaaggt gaacagtgtg aaagagctag acgccttcaa 1201 gctaaaatga tgactaattt ggttatggcc aaggaccgct tacaacttct agagaagatg 1261 caaccagttt tgccattttc caagtcacaa acggacgtct ataatgacag tactaacttg 1321 gcatgccgca atggacatct ccagtcagaa agtggagctg ttccaaaaag aaaagacccc 1381 ttaacacaca ctagtaattc actgcctcgt tcaaaaacag ttatgaaaac tggatctgca 1441 ggcctttcag gccaccatag agcacctagt tacagtggtt tatccatggt ttctggagtg 1501 aaacagggat ctggtcctgc tcctaccact cataagggta ctccgaaaac aaataggaca 1561 aataaacctt ctacccctac aactgctact cgtaagaaaa aagacttgaa gaattttagg 1621 aatgtggaca gcaaccttgc taaccttata atgaatgaaa ttgtggacaa tggaacagct 1681 gttaaatttg atgatatagc tggtcaagac ttggcaaaac aagcattgca agaaattgtt 1741 attcttcctt ctctgaggcc tgagttgttc acagggctta gagctcctgc cagagggctg 1801 ttactctttg gtccacctgg gaatgggaag acaatgctgg ctaaagcagt agctgcagaa 1861 tcgaatgcaa ccttctttaa tataagtgct gcaagtttaa cttcaaaata cgtgggagaa 1921 ggagagaaat tggtgagggc tctttttgct gtggctcgag aacttcaacc ttctataatt 1981 tttatagatg aagttgatag ccttttgtgt gaaagaagag aaggggagca cgatgctagt 2041 agacgcctaa aaactgaatt tctaatagaa tttgatggtg tacagtctgc tggagatgac 2101 agagtacttg taatgggtgc aactaatagg ccacaagagc ttgatgaggc tgttctcagg 2161 cgtttcatca aacgggtata tgtgtcttta ccaaatgagg agacaagact acttttgctt 2221 aaaaatctgt tatgtaaaca aggaagtcca ttgacccaaa aagaactagc acaacttgct 2281 agaatgactg atggatactc aggaagtgac ctaacagctt tggcaaaaga tgcagcactg 2341 ggtcctatcc gagaactaaa accagaacag gtgaagaata tgtctgccag tgagatgaga 2401 aatattcgat tatctgactt cactgaatcc ttgaaaaaaa taaaacgcag cgtcagccct 2461 caaactttag aagcgtacat acgttggaac aaggactttg gagataccac tgtttaaggg 2521 cccggtacct taattaatta aggtaccagg taagtgtacc caattcgccc tatagtgagt 2581 cgtattacaa ttcactcgat cgcccttccc aacagttgcg cagcctgaat ggcgaatgga 2641 gatccaattt ttaagtgtat aatgtgttaa actactgatt ctaattgttt gtgtatttta 2701 gattcacagt cccaaggctc atttcaggcc cctcagtcct cacagtctgt tcatgatcat 2761 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc 2821 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta 2881 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact 2941 gcattctagt tgtggtttgt ccaaactcat caatgtatct taacgcgtaa attgtaagcg 3001 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat 3061 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg 3121 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc 3181 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt 3241 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag 3301 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg 3361 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc 3421 ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc 3481 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct 3541 gataaatgct tcaataatat tgaaaaagga agaatcctga ggcggaaaga accagctgtg 3601 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca 3661 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg 3721 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc 3781 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat 3841 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg 3901 aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat 3961 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga 4021 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc 4081 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga 4141 atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg 4201 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc 4261 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg 4321 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga 4381 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc 4441 tggacgaaga acatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca 4501 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg 4561 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct 4621 atcaggacat agcgttggct acccgtgata ttgctgaaga acttggcggc gaatgggctg 4681 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc 4741 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac 4801 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt 4861 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga 4921 gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg 4981 aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt 5041 tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca 5101 ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga 5161 aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc 5221 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat 5281 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc 5341 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg 5401 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct 5461 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct 5521 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct 5581 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg 5641 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc 5701 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga 5761 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg 5821 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta 5881 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg 5941 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg 6001 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat 6061 taccgccatg cattagttat taatagtaat caattacggg gtcattagtt catagcccat 6121 atatggagtt ccg //