LOCUS       pCMV/3«Box_(GLuc        3769 bp    DNA     circular UNA 07-JUL-2015
DEFINITION  Transient expression of the "TOP1" construct, targeting the GLuc
            reporter, in mammalian cells. CMV/3«Box expression backbone.
ACCESSION   urn.local...1435877562854.102
VERSION     urn.local...1435877562854.102
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,<2592..>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
                     /modified_by="User"
                     /label="CMV_immearly_promoter"
     source          <1463..>2075
                     /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"
     ncRNA           2101..2176
                     /created_by="User"
                     /modified_by="User"
                     /label="sgRNA_Core"
     ncRNA           2177..2201
                     /created_by="User"
                     /modified_by="User"
                     /label="Linker"
     ncRNA           2202..2510
                     /created_by="User"
                     /modified_by="User"
                     /label="P4-P6_[5xPP7_SL]"
     misc_feature    2339..2363
                     /created_by="User"
                     /modified_by="User"
                     /label="PP7_SL1"
     misc_feature    2369..2393
                     /created_by="User"
                     /modified_by="User"
                     /label="PP7_SL2"
     misc_feature    2399..2423
                     /created_by="User"
                     /modified_by="User"
                     /label="PP7_SL3"
     misc_feature    2429..2453
                     /created_by="User"
                     /modified_by="User"
                     /label="PP7_SL4"
     misc_feature    2459..2483
                     /created_by="User"
                     /modified_by="User"
                     /label="PP7_SL5"
     ncRNA           2511..2520
                     /created_by="User"
                     /modified_by="User"
                     /label="U1 Linker"
     terminator      2521..2592
                     /created_by="User"
                     /modified_by="User"
                     /label="U1 snRNA Terminator"
     misc_feature    2521..2534
                     /created_by="User"
                     /modified_by="User"
                     /label="3«Box"
     misc_feature    2541..2563
                     /created_by="User"
                     /modified_by="User"
                     /label="T-Rich_Footprint_Region_(Proudfoot)"
     misc_feature    complement(2609..2625)
                     /created_by="User"
                     /label="M13R Primer From GeneWiz"
                     /note="Geneious type: primer_bind_reverse"
     -10_signal      complement(2658..2663)
                     /note="Plac promoter (drives lacZalpha;
                     counter-clockwise); TATGTT"
                     /label="-10 signal"
     -35_signal      complement(2682..2687)
                     /note="Plac promoter (drives lacZalpha;
                     counter-clockwise); TTTACA"
                     /label="-35 signal"
     protein_bind    complement(2707..2719)
                     /note="CAP protein binding site"
                     /bound_moiety="catabolite activator protein (CAP)"
                     /label="CAP protein binding site protein bind"
     rep_origin      complement(3011..3599)
                     /note="pMB1 origin of replication (counter-clockwise)
                     (RNAII -35 to RNA/DNA switch point)"
                     /label="rep origin"
     misc_RNA        complement(3011..3563)
                     /note="RNAII transcript (complementary strand)"
                     /label="RNA"
     -35_signal      3417..3422
                     /note="RNAI promoter (clockwise); TTGAAG"
                     /label="RNAI promoter (clockwise); TTGAAG -35 signal"
     -10_signal      3439..3444
                     /note="RNAI promoter (clockwise); GCTACA"
                     /label="RNAI promoter (clockwise); GCTACA -10 signal"
     misc_RNA        3453..3560
                     /note="RNAI transcript"
                     /label="RNAI transcript"
     -10_signal      complement(3573..3578)
                     /note="RNAII promoter (counter-clockwise); CGTAAT"
                     /label="-10 signal"
     -35_signal      complement(3594..3599)
                     /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 atgcagttca actttctgga
     2521 gtttcaaaag tagactgtac gctaagggtc atatcttttt ttgtttggtt tgtgtcttgg
     2581 ttggcgtctt aagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     2641 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     2701 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     2761 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     2821 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     2881 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     2941 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     3001 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     3061 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     3121 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     3181 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     3241 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     3301 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     3361 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     3421 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     3481 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     3541 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     3601 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     3661 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     3721 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacag
//
