LOCUS       pU6_(GLuc)_INT[3        3153 bp    DNA     circular UNA 07-JUL-2015
DEFINITION  Transient expression of an "INT" constructÐbearing three PP7
            Stem-loops, targeting the GLuc reporter, in mammalian cells. U6
            promoter.
ACCESSION   urn.local...1435877939353.144
VERSION     urn.local...1435877939353.144
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,<1939..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..1931
                     /created_by="User"
                     /modified_by="User"
                     /label="sgRNA_INT[3xPP7_SL]"
     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..1800
                     /created_by="User"
                     /label="PP7_SL1"
     misc_feature    1804..1828
                     /created_by="User"
                     /label="PP7_SL2"
     misc_feature    1832..1856
                     /created_by="User"
                     /label="PP7_SL3"
     misc_feature    1857..1866
                     /created_by="User"
                     /label="bracketing_helix"
     misc_feature    1867..1931
                     /created_by="User"
                     /label="standard_sgRNA_3«_end"
     terminator      1932..1938
                     /created_by="User"
                     /modified_by="User"
                     /label="Pol III Termination"
     misc_feature    complement(2000..2016)
                     /created_by="User"
                     /label="M13R From Genewiz"
                     /note="Geneious type: primer_bind_reverse"
     -10_signal      complement(2049..2054)
                     /note="Plac promoter (drives lacZalpha;
                     counter-clockwise); TATGTT"
                     /label="-10 signal"
     -35_signal      complement(2073..2078)
                     /note="Plac promoter (drives lacZalpha;
                     counter-clockwise); TTTACA"
                     /label="-35 signal"
     protein_bind    complement(2098..2110)
                     /note="CAP protein binding site"
                     /bound_moiety="catabolite activator protein (CAP)"
                     /label="CAP protein binding site protein bind"
     rep_origin      complement(2395..2983)
                     /note="pMB1 origin of replication (counter-clockwise)
                     (RNAII -35 to RNA/DNA switch point)"
                     /label="rep origin"
     misc_RNA        complement(2395..2947)
                     /note="RNAII transcript (complementary strand)"
                     /label="RNA"
     -35_signal      2801..2806
                     /note="RNAI promoter (clockwise); TTGAAG"
                     /label="RNAI promoter (clockwise); TTGAAG -35 signal"
     -10_signal      2823..2828
                     /note="RNAI promoter (clockwise); GCTACA"
                     /label="RNAI promoter (clockwise); GCTACA -10 signal"
     misc_RNA        2837..2944
                     /note="RNAI transcript"
                     /label="RNAI transcript"
     -10_signal      complement(2957..2962)
                     /note="RNAII promoter (counter-clockwise); CGTAAT"
                     /label="-10 signal"
     -35_signal      complement(2978..2983)
                     /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 acgagggagc agacgatatg gcgtcgctcc
     1801 tctccacgag agcatatggg ctccgtggtc tccagcagac gatatggcgt cgctggctcg
     1861 tattcgcagc atagcaagtt taaataaggc tagtccgtta tcaacttgaa aaagtggcac
     1921 cgagtcggtg ctttttttct taattaagtc tagagtcgac tgtttaaacc tgcaggcatg
     1981 caagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     2041 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     2101 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     2161 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggccgc
     2221 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     2281 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     2341 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     2401 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     2461 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     2521 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     2581 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     2641 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     2701 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     2761 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     2821 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     2881 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     2941 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     3001 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     3061 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     3121 ctaaagtata tatgagtaaa cttggtctga cag
//
