LOCUS       pU6_(GLuc)_TOP1         3303 bp    DNA     circular UNA 07-JUL-2015
DEFINITION  Transient expression of the "TOP1" construct, targeting the GLuc
            reporter, in mammalian cells. U6 promoter.
ACCESSION   urn.local...1435877939364.145
VERSION     urn.local...1435877939364.145
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,<2089..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..1834
                     /created_by="User"
                     /modified_by="User"
                     /label="sgRNA_Core"
     ncRNA           1835..1840
                     /created_by="User"
                     /modified_by="User"
                     /label="Linker"
     ncRNA           1841..2081
                     /created_by="User"
                     /modified_by="User"
                     /label="P4-P6[3xPP7_SL]"
     misc_feature    1976..2000
                     /created_by="User"
                     /label="PP7_SL1"
     misc_feature    2004..2028
                     /created_by="User"
                     /label="PP7_SL2"
     misc_feature    2032..2056
                     /created_by="User"
                     /label="PP7_SL3"
     terminator      2082..2088
                     /created_by="User"
                     /modified_by="User"
                     /label="Pol_III_Termination"
     misc_feature    complement(2150..2166)
                     /created_by="User"
                     /label="M13R From Genewiz"
                     /note="Geneious type: primer_bind_reverse"
     -10_signal      complement(2199..2204)
                     /note="Plac promoter (drives lacZalpha;
                     counter-clockwise); TATGTT"
                     /label="-10 signal"
     -35_signal      complement(2223..2228)
                     /note="Plac promoter (drives lacZalpha;
                     counter-clockwise); TTTACA"
                     /label="-35 signal"
     protein_bind    complement(2248..2260)
                     /note="CAP protein binding site"
                     /bound_moiety="catabolite activator protein (CAP)"
                     /label="CAP protein binding site protein bind"
     rep_origin      complement(2545..3133)
                     /note="pMB1 origin of replication (counter-clockwise)
                     (RNAII -35 to RNA/DNA switch point)"
                     /label="rep origin"
     misc_RNA        complement(2545..3097)
                     /note="RNAII transcript (complementary strand)"
                     /label="RNA"
     -35_signal      2951..2956
                     /note="RNAI promoter (clockwise); TTGAAG"
                     /label="RNAI promoter (clockwise); TTGAAG -35 signal"
     -10_signal      2973..2978
                     /note="RNAI promoter (clockwise); GCTACA"
                     /label="RNAI promoter (clockwise); GCTACA -10 signal"
     misc_RNA        2987..3094
                     /note="RNAI transcript"
                     /label="RNAI transcript"
     -10_signal      complement(3107..3112)
                     /note="RNAII promoter (counter-clockwise); CGTAAT"
                     /label="-10 signal"
     -35_signal      complement(3128..3133)
                     /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 tgctggaaac agcatagcaa gtttaaataa ggctagtccg
     1801 ttatcaactt gaaaaagtgg caccgagtcg gtgcttcatt ggaattgcgg gaaaggggtc
     1861 aacagccgtt cagtaccaag tctcagggga aactttgaga tggccttgca aagggtatgg
     1921 taataagctg acggacatgg tcctaacacg cagccaagtc ctaagtcaac agtctggagc
     1981 agacgatatg gcgtcgctcc tctccacgag agcatatggg ctccgtggtc tccagcagac
     2041 gatatggcgt cgctggtctc tgttgatatg gatgcagttc atttttttct taattaagtc
     2101 tagagtcgac tgtttaaacc tgcaggcatg caagcttggc gtaatcatgg tcatagctgt
     2161 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     2221 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     2281 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     2341 cggggagagg cggtttgcgt attgggccgc ttcctcgctc actgactcgc tgcgctcggt
     2401 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     2461 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     2521 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     2581 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     2641 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     2701 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     2761 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     2821 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     2881 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     2941 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     3001 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     3061 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     3121 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     3181 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     3241 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     3301 cag
//
