LOCUS       pCMV/MASC_(GLuc)        3832 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. CMV/
            MASC expression backbone.
ACCESSION   urn.local...1435877562846.100
VERSION     urn.local...1435877562846.100
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..>1461,<2628..>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        1463..2079
                     /label="CMV_immearly_promoter"
     source          <1463..>2079
                     /organism="pcDNA3.1(+)"
                     /mol_type="other DNA"
                     /label="source pcDNA3.1(+)"
     misc_feature    1542..1829
                     /label="CAG_enhancer"
     primer_bind     1996..2016
                     /modified_by="User"
                     /label="CMV_fwd_primer"
     ncRNA           2080..2100
                     /created_by="User"
                     /modified_by="User"
                     /label="GLuc_gRNA"
     misc_feature    2101..2112
                     /created_by="User"
                     /modified_by="User"
                     /label="sgRNA"
     ncRNA           2101..2330
                     /created_by="User"
                     /label="sgRNA_INT[5xPP7_SL]"
     misc_feature    2113..2122
                     /created_by="User"
                     /modified_by="User"
                     /label="bracketing_helix"
     misc_feature    2123..2147
                     /created_by="User"
                     /label="PP7_SL1"
     misc_feature    2151..2175
                     /created_by="User"
                     /label="PP7_SL2"
     misc_feature    2179..2203
                     /created_by="User"
                     /label="PP7_SL3"
     misc_feature    2207..2231
                     /created_by="User"
                     /label="PP7_SL4"
     misc_feature    2235..2259
                     /created_by="User"
                     /label="PP7_SL5"
     misc_feature    2260..2269
                     /created_by="User"
                     /modified_by="User"
                     /label="bracketing_helix"
     misc_feature    2270..2330
                     /created_by="User"
                     /label="sgRNA"
     ncRNA           2334..2507
                     /created_by="User"
                     /modified_by="User"
                     /label="MALAT1_ENE_and_MASC_RNA"
     misc_feature    2444
                     /created_by="User"
                     /label="RNaseP_Cleavage"
     terminator      2508..2627
                     /label="SV40_PA_terminator"
     source          <2508..>2627
                     /organism="pcDNA3.1(+)"
                     /mol_type="other DNA"
                     /label="source pcDNA3.1(+)"
     misc_feature    complement(2542..2561)
                     /created_by="User"
                     /modified_by="User"
                     /label="SV40pA Rev Primer from Genewiz"
                     /note="Geneious type: primer_bind_reverse"
     primer_bind     2596..2615
                     /modified_by="User"
                     /label="EBV_rev_primer"
     misc_feature    complement(2672..2688)
                     /created_by="User"
                     /label="M13R Primer From GeneWiz"
                     /note="Geneious type: primer_bind_reverse"
     -10_signal      complement(2721..2726)
                     /note="Plac promoter (drives lacZalpha;
                     counter-clockwise); TATGTT"
                     /label="-10 signal"
     -35_signal      complement(2745..2750)
                     /note="Plac promoter (drives lacZalpha;
                     counter-clockwise); TTTACA"
                     /label="-35 signal"
     protein_bind    complement(2770..2782)
                     /note="CAP protein binding site"
                     /bound_moiety="catabolite activator protein (CAP)"
                     /label="CAP protein binding site protein bind"
     rep_origin      complement(3074..3662)
                     /note="pMB1 origin of replication (counter-clockwise)
                     (RNAII -35 to RNA/DNA switch point)"
                     /label="rep origin"
     misc_RNA        complement(3074..3626)
                     /note="RNAII transcript (complementary strand)"
                     /label="RNA"
     -35_signal      3480..3485
                     /note="RNAI promoter (clockwise); TTGAAG"
                     /label="RNAI promoter (clockwise); TTGAAG -35 signal"
     -10_signal      3502..3507
                     /note="RNAI promoter (clockwise); GCTACA"
                     /label="RNAI promoter (clockwise); GCTACA -10 signal"
     misc_RNA        3516..3623
                     /note="RNAI transcript"
                     /label="RNAI transcript"
     -10_signal      complement(3636..3641)
                     /note="RNAII promoter (counter-clockwise); CGTAAT"
                     /label="-10 signal"
     -35_signal      complement(3657..3662)
                     /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 gagctcggta ctacattgat tattgactag ttattaatag taatcaatta
     1501 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
     1561 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
     1621 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
     1681 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
     1741 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
     1801 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
     1861 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
     1921 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
     1981 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
     2041 gagctctctg gctaactaga gaacccactg cttactggcg atctagatac gactcactat
     2101 gttttagagc tacgaatacg agggagcaga cgatatggcg tcgctcctct ccacgagagc
     2161 atatgggctc cgtggtctgc agcagacgat atggcgtcgc tgctctcgta gatgccatat
     2221 ggggcactac gtctccagca gacgatatgg cgtcgctggc tcgtattcgt agcaagttaa
     2281 aataaggcta gtccgttatc aacttgaaaa agtggcaccg agtcggtgct ctcgattcgt
     2341 cagtagggtt gtaaaggttt ttcttttcct gagaaaacaa ccttttgttt tctcaggttt
     2401 tgctttttgg cctttcccta gctttaaaaa aaaaaaagca aaagacgctg gtggctggca
     2461 ctcctggttt ccaggacggg gttcaagtcc ctgcggtgtc tttgcttttg tttattgcag
     2521 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     2581 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttattcg actgtttaaa
     2641 cctgcaggca tgcaagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     2701 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     2761 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     2821 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     2881 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     2941 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     3001 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     3061 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     3121 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     3181 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     3241 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     3301 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     3361 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     3421 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     3481 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     3541 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     3601 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     3661 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     3721 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     3781 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac ag
//
