LOCUS pU6_(GLuc)_TOP1 3303 bp DNA circular UNA 07-JUL-2015 DEFINITION Transient expression of the "TOP1" construct, targeting the GLuc reporter, in mammalian cells. U6 promoter. ACCESSION urn.local...1435877939364.145 VERSION urn.local...1435877939364.145 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" origin 1061^1062 /label="origin" source order(1062..>1487,<2089..1061) /organism="Cloning vector pNEB193" /mol_type="other DNA" /label="source Cloning vector pNEB193" primer_bind 1413..1436 /created_by="User" /label="M13F(-47) From Genewiz" primer_bind 1419..1436 /created_by="User" /label="M13(-41)F From Genewiz" primer_bind 1439..1456 /created_by="User" /label="M13F(-21) From Genewiz" promoter 1488..1727 /created_by="User" /label="U6_Promoter" ncRNA 1728..1748 /created_by="User" /modified_by="User" /label="GLuc_gRNA" ncRNA 1749..1834 /created_by="User" /modified_by="User" /label="sgRNA_Core" ncRNA 1835..1840 /created_by="User" /modified_by="User" /label="Linker" ncRNA 1841..2081 /created_by="User" /modified_by="User" /label="P4-P6[3xPP7_SL]" misc_feature 1976..2000 /created_by="User" /label="PP7_SL1" misc_feature 2004..2028 /created_by="User" /label="PP7_SL2" misc_feature 2032..2056 /created_by="User" /label="PP7_SL3" terminator 2082..2088 /created_by="User" /modified_by="User" /label="Pol_III_Termination" misc_feature complement(2150..2166) /created_by="User" /label="M13R From Genewiz" /note="Geneious type: primer_bind_reverse" -10_signal complement(2199..2204) /note="Plac promoter (drives lacZalpha; counter-clockwise); TATGTT" /label="-10 signal" -35_signal complement(2223..2228) /note="Plac promoter (drives lacZalpha; counter-clockwise); TTTACA" /label="-35 signal" protein_bind complement(2248..2260) /note="CAP protein binding site" /bound_moiety="catabolite activator protein (CAP)" /label="CAP protein binding site protein bind" rep_origin complement(2545..3133) /note="pMB1 origin of replication (counter-clockwise) (RNAII -35 to RNA/DNA switch point)" /label="rep origin" misc_RNA complement(2545..3097) /note="RNAII transcript (complementary strand)" /label="RNA" -35_signal 2951..2956 /note="RNAI promoter (clockwise); TTGAAG" /label="RNAI promoter (clockwise); TTGAAG -35 signal" -10_signal 2973..2978 /note="RNAI promoter (clockwise); GCTACA" /label="RNAI promoter (clockwise); GCTACA -10 signal" misc_RNA 2987..3094 /note="RNAI transcript" /label="RNAI transcript" -10_signal complement(3107..3112) /note="RNAII promoter (counter-clockwise); CGTAAT" /label="-10 signal" -35_signal complement(3128..3133) /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 gccctttcgt ctcgcgcgtt tcggtgatga 1081 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga 1141 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg 1201 gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat 1261 accgcacaga tgcgtaagga gaaaataccg catcaggcgc cattcgccat tcaggctgcg 1321 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg 1381 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg 1441 taaaacgacg gccagtgaat tcgagctcgg tacccggggg cgcgccgttt cccatgattc 1501 cttcatattt gcatatacga tacaaggctg ttagagagat aattggaatt aatttgactg 1561 taaacacaaa gatattagta caaaatacgt gacgtagaaa gtaataattt cttgggtagt 1621 ttgcagtttt aaaattatgt tttaaaatgg actatcatat gcttaccgta acttgaaagt 1681 atttcgattt cttggcttta tatatcttgt ggaaaggacg aaacaccgat ctagatacga 1741 ctcactatgt ttaagagcta tgctggaaac agcatagcaa gtttaaataa ggctagtccg 1801 ttatcaactt gaaaaagtgg caccgagtcg gtgcttcatt ggaattgcgg gaaaggggtc 1861 aacagccgtt cagtaccaag tctcagggga aactttgaga tggccttgca aagggtatgg 1921 taataagctg acggacatgg tcctaacacg cagccaagtc ctaagtcaac agtctggagc 1981 agacgatatg gcgtcgctcc tctccacgag agcatatggg ctccgtggtc tccagcagac 2041 gatatggcgt cgctggtctc tgttgatatg gatgcagttc atttttttct taattaagtc 2101 tagagtcgac tgtttaaacc tgcaggcatg caagcttggc gtaatcatgg tcatagctgt 2161 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa 2221 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac 2281 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg 2341 cggggagagg cggtttgcgt attgggccgc ttcctcgctc actgactcgc tgcgctcggt 2401 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga 2461 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg 2521 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa 2581 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt 2641 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct 2701 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct 2761 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc 2821 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt 2881 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc 2941 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat 3001 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa 3061 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa 3121 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga 3181 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct 3241 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga 3301 cag //