LOCUS       pCMV/MASC_(GLuc)        4010 bp    DNA     circular UNA 07-JUL-2015
DEFINITION  Transient expression of the "TOP1" construct, targeting the GLuc
            reporter, in mammalian cells. CMV/MASC expression backbone.
ACCESSION   urn.local...1435877562843.99
VERSION     urn.local...1435877562843.99
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,<2806..>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"
                     /label="GLuc_gRNA"
     ncRNA           2101..2176
                     /created_by="User"
                     /label="sgRNA_Core"
     ncRNA           2177..2201
                     /created_by="User"
                     /label="Linker"
     ncRNA           2202..2509
                     /created_by="User"
                     /label="P4-P6_[5xPP7_SL]"
     misc_feature    2339..2363
                     /created_by="User"
                     /label="PP7_SL1"
     misc_feature    2369..2393
                     /created_by="User"
                     /label="PP7_SL2"
     misc_feature    2399..2423
                     /created_by="User"
                     /label="PP7_SL3"
     misc_feature    2429..2453
                     /created_by="User"
                     /label="PP7_SL4"
     misc_feature    2459..2483
                     /created_by="User"
                     /label="PP7_SL5"
     ncRNA           2512..2685
                     /created_by="User"
                     /modified_by="User"
                     /label="MALAT1_ENE_and_MASC_RNA"
     misc_feature    2622
                     /created_by="User"
                     /label="RNAseP_Cleavage"
     terminator      2686..2805
                     /label="SV40_PA_terminator"
     source          <2686..>2805
                     /organism="pcDNA3.1(+)"
                     /mol_type="other DNA"
                     /label="source pcDNA3.1(+)"
     misc_feature    complement(2720..2739)
                     /created_by="User"
                     /modified_by="User"
                     /label="SV40pA Rev Primer from Genewiz"
                     /note="Geneious type: primer_bind_reverse"
     primer_bind     2774..2793
                     /modified_by="User"
                     /label="EBV_rev_primer"
     misc_feature    complement(2850..2866)
                     /created_by="User"
                     /label="M13R Primer From GeneWiz"
                     /note="Geneious type: primer_bind_reverse"
     -10_signal      complement(2899..2904)
                     /note="Plac promoter (drives lacZalpha;
                     counter-clockwise); TATGTT"
                     /label="-10 signal"
     -35_signal      complement(2923..2928)
                     /note="Plac promoter (drives lacZalpha;
                     counter-clockwise); TTTACA"
                     /label="-35 signal"
     protein_bind    complement(2948..2960)
                     /note="CAP protein binding site"
                     /bound_moiety="catabolite activator protein (CAP)"
                     /label="CAP protein binding site protein bind"
     rep_origin      complement(3252..3840)
                     /note="pMB1 origin of replication (counter-clockwise)
                     (RNAII -35 to RNA/DNA switch point)"
                     /label="rep origin"
     misc_RNA        complement(3252..3804)
                     /note="RNAII transcript (complementary strand)"
                     /label="RNA"
     -35_signal      3658..3663
                     /note="RNAI promoter (clockwise); TTGAAG"
                     /label="RNAI promoter (clockwise); TTGAAG -35 signal"
     -10_signal      3680..3685
                     /note="RNAI promoter (clockwise); GCTACA"
                     /label="RNAI promoter (clockwise); GCTACA -10 signal"
     misc_RNA        3694..3801
                     /note="RNAI transcript"
                     /label="RNAI transcript"
     -10_signal      complement(3814..3819)
                     /note="RNAII promoter (counter-clockwise); CGTAAT"
                     /label="-10 signal"
     -35_signal      complement(3835..3840)
                     /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 tagaaatagc aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt
     2161 ggcaccgagt cggtgctctt caacatccct cctctaaacc cggaattgcg ggaaaggggt
     2221 caacagccgt tcagtaccaa gtctcagggg aaactttgag atggccttgc aaagggtatg
     2281 gtaataagct gacggacatg gtcctaacac gcagccaagt cctaagtcaa cagtttttgg
     2341 agcagacgat atggcgtcgc tcctttttcc agcagagcat atgggctccg tggtttttcc
     2401 agcagacgat atggcgtcgc tggtttttgg agcagagcat atgggctcgc tcctttttgc
     2461 agcagacgat atggcgtcgc tgctttttct gttgatatgg atgcagttct cgattcgtca
     2521 gtagggttgt aaaggttttt cttttcctga gaaaacaacc ttttgttttc tcaggttttg
     2581 ctttttggcc tttccctagc tttaaaaaaa aaaaagcaaa agacgctggt ggctggcact
     2641 cctggtttcc aggacggggt tcaagtccct gcggtgtctt tgcttttgtt tattgcagct
     2701 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     2761 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttattcgac tgtttaaacc
     2821 tgcaggcatg caagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     2881 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     2941 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     3001 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     3061 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     3121 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     3181 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     3241 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     3301 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     3361 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     3421 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     3481 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     3541 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     3601 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     3661 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     3721 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     3781 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     3841 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     3901 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     3961 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
//
