LOCUS pU6_(GLuc)_INT[G 3154 bp DNA circular UNA 07-JUL-2015 DEFINITION Transient expression in mammalian cells of an "INT" constructÐbearing the GFP aptamer, targeting the GLuc reporter. U6 promoter. ACCESSION urn.local...1435877939316.137 VERSION urn.local...1435877939316.137 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,<1940..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..1932 /created_by="User" /modified_by="User" /label="sgRNA_INT[GFP_Aptamer]" misc_feature 1749..1765 /created_by="User" /label="standard_sgRNA_5«_end" misc_feature 1766..1775 /created_by="User" /label="bracketing_helix" misc_feature 1776..1857 /created_by="User" /label="GFP Aptamer" misc_feature 1858..1867 /created_by="User" /label="bracketing_helix" misc_feature 1868..1932 /created_by="User" /label="standard_sgRNA_3«_end" terminator 1933..1939 /created_by="User" /modified_by="User" /label="Pol_III_Terminator" misc_feature complement(2001..2017) /created_by="User" /label="M13R From Genewiz" /note="Geneious type: primer_bind_reverse" -10_signal complement(2050..2055) /note="Plac promoter (drives lacZalpha; counter-clockwise); TATGTT" /label="-10 signal" -35_signal complement(2074..2079) /note="Plac promoter (drives lacZalpha; counter-clockwise); TTTACA" /label="-35 signal" protein_bind complement(2099..2111) /note="CAP protein binding site" /bound_moiety="catabolite activator protein (CAP)" /label="CAP protein binding site protein bind" rep_origin complement(2396..2984) /note="pMB1 origin of replication (counter-clockwise) (RNAII -35 to RNA/DNA switch point)" /label="rep origin" misc_RNA complement(2396..2948) /note="RNAII transcript (complementary strand)" /label="RNA" -35_signal 2802..2807 /note="RNAI promoter (clockwise); TTGAAG" /label="RNAI promoter (clockwise); TTGAAG -35 signal" -10_signal 2824..2829 /note="RNAI promoter (clockwise); GCTACA" /label="RNAI promoter (clockwise); GCTACA -10 signal" misc_RNA 2838..2945 /note="RNAI transcript" /label="RNAI transcript" -10_signal complement(2958..2963) /note="RNAII promoter (counter-clockwise); CGTAAT" /label="-10 signal" -35_signal complement(2979..2984) /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 tgctgcgaat acgaggcttc tggactgcga tgggagcacg 1801 aaacgtcgtg gcgcaattgg gtggggaaag tccttaaaag agggccacca cagaagcctc 1861 gtattcgcag catagcaagt ttaaataagg ctagtccgtt atcaacttga aaaagtggca 1921 ccgagtcggt gctttttttc ttaattaagt ctagagtcga ctgtttaaac ctgcaggcat 1981 gcaagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca 2041 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg 2101 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg 2161 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggccg 2221 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc 2281 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt 2341 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc 2401 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa 2461 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc 2521 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg 2581 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc 2641 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc 2701 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca 2761 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact 2821 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg 2881 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt 2941 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct 3001 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga 3061 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa 3121 tctaaagtat atatgagtaa acttggtctg acag //