LOCUS pU1/Sm/3«Box_(GL 3578 bp DNA circular UNA 07-JUL-2015 DEFINITION Transient expression of the "TOP1" construct, targeting the GLuc reporter, in mammalian cells. U1Pro/SmBox/3«Box expression backbone. ACCESSION urn.local...1435877562864.105 VERSION urn.local...1435877562864.105 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..>1450,<2401..>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 1451..1843 /created_by="User" /label="hU1_Pro" ncRNA 1844..1865 /created_by="User" /modified_by="User" /label="GLuc_gRNA (with additional 5«A)" ncRNA 1866..1951 /created_by="User" /modified_by="User" /label="sgRNA_Core" ncRNA 1952..1957 /created_by="User" /modified_by="User" /label="Linker" ncRNA 1958..2198 /created_by="User" /modified_by="User" /label="P4-P6_[3xPP7_SL]" misc_feature 2093..2117 /created_by="User" /label="PP7_SL1" misc_feature 2121..2145 /created_by="User" /label="PP7_SL2" misc_feature 2149..2173 /created_by="User" /label="PP7_SL3" ncRNA 2199..2228 /created_by="User" /modified_by="User" /label="""Linker""_stem_loop" ncRNA 2229..2319 /created_by="User" /modified_by="User" /label="U2_snRNA_sm_Box" ncRNA 2320..2329 /created_by="User" /modified_by="User" /label="U1 Linker" terminator 2330..2401 /created_by="User" /modified_by="User" /label="U1 snRNA Terminator" misc_feature 2330..2343 /created_by="User" /modified_by="User" /label="3«Box" misc_feature 2350..2372 /created_by="User" /modified_by="User" /label="T-Rich_Footprint_Region_(Proudfoot)" misc_feature complement(2418..2434) /created_by="User" /label="M13R Primer From GeneWiz" /note="Geneious type: primer_bind_reverse" -10_signal complement(2467..2472) /note="Plac promoter (drives lacZalpha; counter-clockwise); TATGTT" /label="-10 signal" -35_signal complement(2491..2496) /note="Plac promoter (drives lacZalpha; counter-clockwise); TTTACA" /label="-35 signal" protein_bind complement(2516..2528) /note="CAP protein binding site" /bound_moiety="catabolite activator protein (CAP)" /label="CAP protein binding site protein bind" rep_origin complement(2820..3408) /note="pMB1 origin of replication (counter-clockwise) (RNAII -35 to RNA/DNA switch point)" /label="rep origin" misc_RNA complement(2820..3372) /note="RNAII transcript (complementary strand)" /label="RNA" -35_signal 3226..3231 /note="RNAI promoter (clockwise); TTGAAG" /label="RNAI promoter (clockwise); TTGAAG -35 signal" -10_signal 3248..3253 /note="RNAI promoter (clockwise); GCTACA" /label="RNAI promoter (clockwise); GCTACA -10 signal" misc_RNA 3262..3369 /note="RNAI transcript" /label="RNAI transcript" -10_signal complement(3382..3387) /note="RNAII promoter (counter-clockwise); CGTAAT" /label="-10 signal" -35_signal complement(3403..3408) /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 ctaaggacca gcttctttgg gagagaacag acgcaggggc gggagggaaa 1501 aagggagagg cagacgtcac ttccccttgg cggctctggc agcagattgg tcggttgagt 1561 ggcagaaagg cagacgggga ctgggcaagg cactgtcggt gacatcacgg acagggcgac 1621 ttctatgtag atgaggcagc gcagaggctg ctgcttcgcc acttgctgct tcaccacgaa 1681 ggagttcccg tgccctggga gcgggttcag gaccgctgat cggaagtgag aatcccagct 1741 gtgtgtcagg gctggaaagg gctcgggagt gcgcggggca agtgaccgtg tgtgtaaaga 1801 gtgaggcgta tgaggctgtg tcggggcaga ggcccaagat ctcagatcta gatacgactc 1861 actatgttta agagctatgc tggaaacagc atagcaagtt taaataaggc tagtccgtta 1921 tcaacttgaa aaagtggcac cgagtcggtg cttcattgga attgcgggaa aggggtcaac 1981 agccgttcag taccaagtct caggggaaac tttgagatgg ccttgcaaag ggtatggtaa 2041 taagctgacg gacatggtcc taacacgcag ccaagtccta agtcaacagt ctggagcaga 2101 cgatatggcg tcgctcctct ccacgagagc atatgggctc cgtggtctcc agcagacgat 2161 atggcgtcgc tggtctctgt tgatatggat gcagttcaca gcaagttcag agaaatctga 2221 acttgctgga tttttggagc agggagatgg aataggagct tgctccgtcc actccacgca 2281 tcgacctggt attgcagtac ctccaggaac ggtgcaccca ctttctggag tttcaaaagt 2341 agactgtacg ctaagggtca tatctttttt tgtttggttt gtgtcttggt tggcgtctta 2401 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt 2461 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc 2521 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc 2581 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct 2641 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca 2701 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac 2761 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt 2821 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg 2881 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc 2941 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc 3001 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc 3061 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac 3121 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt 3181 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct 3241 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc 3301 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt 3361 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg 3421 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc 3481 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa 3541 tcaatctaaa gtatatatga gtaaacttgg tctgacag //