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
//
