LOCUS       pU1/Sm/3«Box_(GL        3428 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.
            U1Pro/SmBox/3«Box expression backbone.
ACCESSION   urn.local...1435877562861.104
VERSION     urn.local...1435877562861.104
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,<2251..>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..2048
                     /created_by="User"
                     /modified_by="User"
                     /label="sgRNA_INT[3xPP7_SL]"
     misc_feature    1866..1882
                     /created_by="User"
                     /label="sgRNA"
     misc_feature    1883..1892
                     /created_by="User"
                     /label="bracketing_helix"
     misc_feature    1893..1917
                     /created_by="User"
                     /label="PP7_SL1"
     misc_feature    1921..1945
                     /created_by="User"
                     /label="PP7_SL2"
     misc_feature    1949..1973
                     /created_by="User"
                     /modified_by="User"
                     /label="PP7_SL3"
     misc_feature    1974..1983
                     /created_by="User"
                     /label="bracketing_helix"
     misc_feature    1984..2048
                     /created_by="User"
                     /label="sgRNA"
     ncRNA           2049..2078
                     /created_by="User"
                     /modified_by="User"
                     /label="""Linker"" stem-loop"
     ncRNA           2079..2169
                     /created_by="User"
                     /modified_by="User"
                     /label="U2_snRNA_sm_Box"
     ncRNA           2170..2179
                     /created_by="User"
                     /modified_by="User"
                     /label="U1 Linker"
     terminator      2180..2251
                     /created_by="User"
                     /modified_by="User"
                     /label="U1 snRNA Terminator"
     misc_feature    2180..2193
                     /created_by="User"
                     /modified_by="User"
                     /label="3«Box"
     misc_feature    2200..2222
                     /created_by="User"
                     /modified_by="User"
                     /label="T-Rich_Footprint_Region_(Proudfoot)"
     misc_feature    complement(2268..2284)
                     /created_by="User"
                     /label="M13R Primer From GeneWiz"
                     /note="Geneious type: primer_bind_reverse"
     -10_signal      complement(2317..2322)
                     /note="Plac promoter (drives lacZalpha;
                     counter-clockwise); TATGTT"
                     /label="-10 signal"
     -35_signal      complement(2341..2346)
                     /note="Plac promoter (drives lacZalpha;
                     counter-clockwise); TTTACA"
                     /label="-35 signal"
     protein_bind    complement(2366..2378)
                     /note="CAP protein binding site"
                     /bound_moiety="catabolite activator protein (CAP)"
                     /label="CAP protein binding site protein bind"
     rep_origin      complement(2670..3258)
                     /note="pMB1 origin of replication (counter-clockwise)
                     (RNAII -35 to RNA/DNA switch point)"
                     /label="rep origin"
     misc_RNA        complement(2670..3222)
                     /note="RNAII transcript (complementary strand)"
                     /label="RNA"
     -35_signal      3076..3081
                     /note="RNAI promoter (clockwise); TTGAAG"
                     /label="RNAI promoter (clockwise); TTGAAG -35 signal"
     -10_signal      3098..3103
                     /note="RNAI promoter (clockwise); GCTACA"
                     /label="RNAI promoter (clockwise); GCTACA -10 signal"
     misc_RNA        3112..3219
                     /note="RNAI transcript"
                     /label="RNAI transcript"
     -10_signal      complement(3232..3237)
                     /note="RNAII promoter (counter-clockwise); CGTAAT"
                     /label="-10 signal"
     -35_signal      complement(3253..3258)
                     /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 tgcgaatacg agggagcaga cgatatggcg tcgctcctct
     1921 ccacgagagc atatgggctc cgtggtctcc agcagacgat atggcgtcgc tggctcgtat
     1981 tcgcagcata gcaagtttaa ataaggctag tccgttatca acttgaaaaa gtggcaccga
     2041 gtcggtgcca gcaagttcag agaaatctga acttgctgga tttttggagc agggagatgg
     2101 aataggagct tgctccgtcc actccacgca tcgacctggt attgcagtac ctccaggaac
     2161 ggtgcaccca ctttctggag tttcaaaagt agactgtacg ctaagggtca tatctttttt
     2221 tgtttggttt gtgtcttggt tggcgtctta agcttggcgt aatcatggtc atagctgttt
     2281 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     2341 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     2401 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     2461 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     2521 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     2581 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     2641 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     2701 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     2761 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     2821 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     2881 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     2941 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     3001 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     3061 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     3121 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     3181 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     3241 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     3301 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     3361 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     3421 tctgacag
//
