LOCUS       pU6_(GLuc)_INT[B        3201 bp    DNA     circular UNA 07-JUL-2015
DEFINITION  Transient expression in mammalian cells of an "INT"
            constructÐbearing the "Bunch of Baby Spinach" (or "BoBS")
            construct, targeting the GLuc reporter. U6 promoter.
ACCESSION   urn.local...1435877939309.135
VERSION     urn.local...1435877939309.135
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,<1987..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..1979
                     /created_by="User"
                     /modified_by="User"
                     /label="sgRNA_INT[BoBS]"
     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..1904
                     /created_by="User"
                     /modified_by="User"
                     /label="""Bunch_of_Baby_Spinach"""
     misc_feature    1905..1914
                     /created_by="User"
                     /label="bracketing_helix"
     misc_feature    1915..1979
                     /created_by="User"
                     /label="standard_sgRNA_3«_end"
     terminator      1980..1986
                     /created_by="User"
                     /modified_by="User"
                     /label="Pol_III_Terminator"
     misc_feature    complement(2048..2064)
                     /created_by="User"
                     /label="M13R From Genewiz"
                     /note="Geneious type: primer_bind_reverse"
     -10_signal      complement(2097..2102)
                     /note="Plac promoter (drives lacZalpha;
                     counter-clockwise); TATGTT"
                     /label="-10 signal"
     -35_signal      complement(2121..2126)
                     /note="Plac promoter (drives lacZalpha;
                     counter-clockwise); TTTACA"
                     /label="-35 signal"
     protein_bind    complement(2146..2158)
                     /note="CAP protein binding site"
                     /bound_moiety="catabolite activator protein (CAP)"
                     /label="CAP protein binding site protein bind"
     rep_origin      complement(2443..3031)
                     /note="pMB1 origin of replication (counter-clockwise)
                     (RNAII -35 to RNA/DNA switch point)"
                     /label="rep origin"
     misc_RNA        complement(2443..2995)
                     /note="RNAII transcript (complementary strand)"
                     /label="RNA"
     -35_signal      2849..2854
                     /note="RNAI promoter (clockwise); TTGAAG"
                     /label="RNAI promoter (clockwise); TTGAAG -35 signal"
     -10_signal      2871..2876
                     /note="RNAI promoter (clockwise); GCTACA"
                     /label="RNAI promoter (clockwise); GCTACA -10 signal"
     misc_RNA        2885..2992
                     /note="RNAI transcript"
                     /label="RNAI transcript"
     -10_signal      complement(3005..3010)
                     /note="RNAII promoter (counter-clockwise); CGTAAT"
                     /label="-10 signal"
     -35_signal      complement(3026..3031)
                     /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 acgagaagga cgggtccgga cgcaaggacg
     1801 ggtccgaccg aaaggacggg tccaatggtg gaaacaccat tgttgagtag agtgtgagtc
     1861 ggtcgttgag tagagtgtga ggcgtccgtt gagtagagtg tgagctcgta ttcgcagcat
     1921 agcaagttta aataaggcta gtccgttatc aacttgaaaa agtggcaccg agtcggtgct
     1981 ttttttctta attaagtcta gagtcgactg tttaaacctg caggcatgca agcttggcgt
     2041 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     2101 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     2161 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     2221 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggccgctt cctcgctcac
     2281 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     2341 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     2401 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     2461 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     2521 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     2581 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     2641 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     2701 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     2761 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     2821 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     2881 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     2941 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     3001 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     3061 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     3121 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     3181 tgagtaaact tggtctgaca g
//
