LOCUS       pU6_(GLuc)_INT[3        3171 bp    DNA     circular UNA 07-JUL-2015
DEFINITION  Transient expression in mammalian cells of an "INT"
            constructÐbearing three kink-turns, targeting the GLuc reporter. U6
            promoter.
ACCESSION   urn.local...1435877939342.142
VERSION     urn.local...1435877939342.142
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,<1957..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..1949
                     /created_by="User"
                     /modified_by="User"
                     /label="sgRNA_INT[3x_K-T]"
     misc_feature    1749..1765
                     /created_by="User"
                     /label="standard_sgRNA_5«_end"
     misc_feature    1766..1775
                     /created_by="User"
                     /label="bracketing_helix"
     misc_feature    1779..1807
                     /created_by="User"
                     /label="Kink-Turn_1"
     misc_feature    1811..1839
                     /created_by="User"
                     /label="Kink-Turn_2"
     misc_feature    1843..1871
                     /created_by="User"
                     /label="Kink-Turn_3"
     misc_feature    1875..1884
                     /created_by="User"
                     /label="bracketing_helix"
     misc_feature    1885..1949
                     /created_by="User"
                     /label="standard_sgRNA_3«_end"
     terminator      1950..1956
                     /created_by="User"
                     /modified_by="User"
                     /label="Pol III Termination"
     misc_feature    complement(2018..2034)
                     /created_by="User"
                     /label="M13R From Genewiz"
                     /note="Geneious type: primer_bind_reverse"
     -10_signal      complement(2067..2072)
                     /note="Plac promoter (drives lacZalpha;
                     counter-clockwise); TATGTT"
                     /label="-10 signal"
     -35_signal      complement(2091..2096)
                     /note="Plac promoter (drives lacZalpha;
                     counter-clockwise); TTTACA"
                     /label="-35 signal"
     protein_bind    complement(2116..2128)
                     /note="CAP protein binding site"
                     /bound_moiety="catabolite activator protein (CAP)"
                     /label="CAP protein binding site protein bind"
     rep_origin      complement(2413..3001)
                     /note="pMB1 origin of replication (counter-clockwise)
                     (RNAII -35 to RNA/DNA switch point)"
                     /label="rep origin"
     misc_RNA        complement(2413..2965)
                     /note="RNAII transcript (complementary strand)"
                     /label="RNA"
     -35_signal      2819..2824
                     /note="RNAI promoter (clockwise); TTGAAG"
                     /label="RNAI promoter (clockwise); TTGAAG -35 signal"
     -10_signal      2841..2846
                     /note="RNAI promoter (clockwise); GCTACA"
                     /label="RNAI promoter (clockwise); GCTACA -10 signal"
     misc_RNA        2855..2962
                     /note="RNAI transcript"
                     /label="RNAI transcript"
     -10_signal      complement(2975..2980)
                     /note="RNAII promoter (counter-clockwise); CGTAAT"
                     /label="-10 signal"
     -35_signal      complement(2996..3001)
                     /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 acgagtctgc tcccgtgatg gcgaaagcct
     1801 gaggagctct ctggccgtga tggcgaaagc ctgagccagt ctctgcccgt gatggcgaaa
     1861 gcctgaggca gtctctcgta ttcgcagcat agcaagttta aataaggcta gtccgttatc
     1921 aacttgaaaa agtggcaccg agtcggtgct ttttttctta attaagtcta gagtcgactg
     1981 tttaaacctg caggcatgca agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     2041 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     2101 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     2161 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     2221 gtttgcgtat tgggccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     2281 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     2341 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     2401 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     2461 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     2521 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     2581 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     2641 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     2701 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     2761 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     2821 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     2881 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     2941 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     3001 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     3061 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     3121 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca g
//
