LOCUS pCMV/MASC_(GLuc) 4010 bp DNA circular UNA 07-JUL-2015 DEFINITION Transient expression of the "TOP1" construct, targeting the GLuc reporter, in mammalian cells. CMV/MASC expression backbone. ACCESSION urn.local...1435877562843.99 VERSION urn.local...1435877562843.99 KEYWORDS . SOURCE ORGANISM . FEATURES Location/Qualifiers CDS complement(1..861) /gene="bla" /note="ampR (confers resistance to ampicillin)" /codon_start=1 /product="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA EIGASLIKHW" /label="bla CDS" gene complement(1..861) /gene="bla" /label="bla gene" sig_peptide complement(793..861) /gene="bla" /note="required for secretion to the periplasm; cleaved off to form the mature beta-lactamase protein." /label="bla signal peptide" -10_signal complement(905..910) /note="bla promoter (counter-clockwise); GAGACA; transcript start 2548 (complementary strand)" /label="-10 signal" -35_signal complement(926..931) /note="bla promoter (counter-clockwise); TTCAAA" /label="-35 signal" source order(<1058..>1461,<2806..>1057) /organism="Cloning vector pNEB193" /mol_type="other DNA" /label="source Cloning vector pNEB193" primer_bind 1401..1424 /created_by="User" /label="M13F (-47) Primer from GeneWiz" primer_bind 1427..1444 /created_by="User" /label="M13F(-21) Primer From GeneWiz" promoter 1463..2079 /label="CMV_immearly_promoter" source <1463..>2079 /organism="pcDNA3.1(+)" /mol_type="other DNA" /label="source pcDNA3.1(+)" misc_feature 1542..1829 /label="CAG_enhancer" primer_bind 1996..2016 /modified_by="User" /label="CMV_fwd_primer" ncRNA 2080..2100 /created_by="User" /label="GLuc_gRNA" ncRNA 2101..2176 /created_by="User" /label="sgRNA_Core" ncRNA 2177..2201 /created_by="User" /label="Linker" ncRNA 2202..2509 /created_by="User" /label="P4-P6_[5xPP7_SL]" misc_feature 2339..2363 /created_by="User" /label="PP7_SL1" misc_feature 2369..2393 /created_by="User" /label="PP7_SL2" misc_feature 2399..2423 /created_by="User" /label="PP7_SL3" misc_feature 2429..2453 /created_by="User" /label="PP7_SL4" misc_feature 2459..2483 /created_by="User" /label="PP7_SL5" ncRNA 2512..2685 /created_by="User" /modified_by="User" /label="MALAT1_ENE_and_MASC_RNA" misc_feature 2622 /created_by="User" /label="RNAseP_Cleavage" terminator 2686..2805 /label="SV40_PA_terminator" source <2686..>2805 /organism="pcDNA3.1(+)" /mol_type="other DNA" /label="source pcDNA3.1(+)" misc_feature complement(2720..2739) /created_by="User" /modified_by="User" /label="SV40pA Rev Primer from Genewiz" /note="Geneious type: primer_bind_reverse" primer_bind 2774..2793 /modified_by="User" /label="EBV_rev_primer" misc_feature complement(2850..2866) /created_by="User" /label="M13R Primer From GeneWiz" /note="Geneious type: primer_bind_reverse" -10_signal complement(2899..2904) /note="Plac promoter (drives lacZalpha; counter-clockwise); TATGTT" /label="-10 signal" -35_signal complement(2923..2928) /note="Plac promoter (drives lacZalpha; counter-clockwise); TTTACA" /label="-35 signal" protein_bind complement(2948..2960) /note="CAP protein binding site" /bound_moiety="catabolite activator protein (CAP)" /label="CAP protein binding site protein bind" rep_origin complement(3252..3840) /note="pMB1 origin of replication (counter-clockwise) (RNAII -35 to RNA/DNA switch point)" /label="rep origin" misc_RNA complement(3252..3804) /note="RNAII transcript (complementary strand)" /label="RNA" -35_signal 3658..3663 /note="RNAI promoter (clockwise); TTGAAG" /label="RNAI promoter (clockwise); TTGAAG -35 signal" -10_signal 3680..3685 /note="RNAI promoter (clockwise); GCTACA" /label="RNAI promoter (clockwise); GCTACA -10 signal" misc_RNA 3694..3801 /note="RNAI transcript" /label="RNAI transcript" -10_signal complement(3814..3819) /note="RNAII promoter (counter-clockwise); CGTAAT" /label="-10 signal" -35_signal complement(3835..3840) /note="RNAII promoter (counter-clockwise); TTGAGA" /label="-35 signal" ORIGIN 1 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat 61 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc 121 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa 181 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca 241 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa 301 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt 361 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc 421 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact 481 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc 541 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg 601 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct 661 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc 721 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag 781 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac 841 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg 901 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt 961 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac 1021 attaacctat aaaaataggc gtatcacgag gccctttgcg cgtttcggtg atgacggtga 1081 aaacctctga cacatgcagc tcccggtcac agcttgtctg taagcggatg ccgggagcag 1141 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc 1201 ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg 1261 cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggga 1321 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc 1381 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc 1441 cagtgaattc gagctcggta ctacattgat tattgactag ttattaatag taatcaatta 1501 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg 1561 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc 1621 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa 1681 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca 1741 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta 1801 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt 1861 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg 1921 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca 1981 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca 2041 gagctctctg gctaactaga gaacccactg cttactggcg atctagatac gactcactat 2101 gttttagagc tagaaatagc aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt 2161 ggcaccgagt cggtgctctt caacatccct cctctaaacc cggaattgcg ggaaaggggt 2221 caacagccgt tcagtaccaa gtctcagggg aaactttgag atggccttgc aaagggtatg 2281 gtaataagct gacggacatg gtcctaacac gcagccaagt cctaagtcaa cagtttttgg 2341 agcagacgat atggcgtcgc tcctttttcc agcagagcat atgggctccg tggtttttcc 2401 agcagacgat atggcgtcgc tggtttttgg agcagagcat atgggctcgc tcctttttgc 2461 agcagacgat atggcgtcgc tgctttttct gttgatatgg atgcagttct cgattcgtca 2521 gtagggttgt aaaggttttt cttttcctga gaaaacaacc ttttgttttc tcaggttttg 2581 ctttttggcc tttccctagc tttaaaaaaa aaaaagcaaa agacgctggt ggctggcact 2641 cctggtttcc aggacggggt tcaagtccct gcggtgtctt tgcttttgtt tattgcagct 2701 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca 2761 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttattcgac tgtttaaacc 2821 tgcaggcatg caagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat 2881 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc 2941 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga 3001 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt 3061 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg 3121 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac 3181 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg 3241 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca 3301 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc 3361 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc 3421 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag 3481 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc 3541 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca 3601 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg 3661 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg 3721 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct 3781 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa 3841 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa 3901 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa 3961 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag //