LOCUS       pUBC_HA_2xhPP7~V        4990 bp    DNA     circular UNA 07-JUL-2015
DEFINITION  Transient expression of humanized PP7(?FG), fused to the VP64
            transcription activator, in human cells, under a UBC promoter.
ACCESSION   urn.local...1435874057537.24
VERSION     urn.local...1435874057537.24
KEYWORDS    .
SOURCE      
  ORGANISM  
            .
FEATURES             Location/Qualifiers
     gene            complement(1..861)
                     /gene="bla"
                     /label="bla gene"
     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"
     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          1050^1051
                     /label="origin"
     source          order(<1051..>1317,<3790..>4230,<4231..>1050)
                     /organism="Cloning vector pNEB193"
                     /mol_type="other DNA"
                     /label="source Cloning vector pNEB193"
     primer_bind     1257..1280
                     /created_by="User"
                     /modified_by="User"
                     /label="M13(-47)F Primer from Genewiz"
     primer_bind     1283..1300
                     /created_by="User"
                     /modified_by="User"
                     /label="M13(-21)F Primer from Genewiz"
     Promoter        1318..2522
                     /created_by="User"
                     /label="human_UbC_Promoter"
     misc_feature    2539..2548
                     /created_by="User"
                     /label="Kozak_Sequence"
     CDS             2545..3042
                     /created_by="User"
                     /modified_by="User"
                     /label="3xNLS_HA_PP7([delta]FG)"
     sig_peptide     2548..2571
                     /created_by="User"
                     /label="NLS"
     sig_peptide     2572..2595
                     /created_by="User"
                     /label="NLS"
     sig_peptide     2596..2619
                     /created_by="User"
                     /label="NLS"
     misc_feature    2623..2649
                     /created_by="User"
                     /label="HA_Tag"
     CDS             3043..3063
                     /created_by="User"
                     /modified_by="User"
                     /label="Linker"
     CDS             3064..3438
                     /created_by="User"
                     /modified_by="User"
                     /label="PP7([delta]FG)"
     CDS             3439..3453
                     /created_by="User"
                     /modified_by="User"
                     /label="Linker"
     CDS             3454..3627
                     /created_by="User"
                     /label="VP64_Transcription_Activator"
     terminator      3634..3636
                     /created_by="User"
                     /label="stop"
     terminator      3638..3640
                     /created_by="User"
                     /label="stop"
     terminator      3642..3644
                     /created_by="User"
                     /label="stop"
     terminator      3670..3789
                     /created_by="User"
                     /label="SV40_pA_terminator"
     misc_feature    complement(3704..3723)
                     /created_by="User"
                     /label="SV40pA_Primer_from_Genewiz"
                     /note="Geneious type: primer_bind_reverse"
     primer_bind     3758..3777
                     /created_by="User"
                     /label="EBV_Rev_Primer"
     misc_feature    complement(3830..3846)
                     /created_by="User"
                     /label="M13R Primer from GeneWiz"
                     /note="Geneious type: primer_bind_reverse"
     -10_signal      complement(3879..3884)
                     /note="Plac promoter (drives lacZalpha;
                     counter-clockwise); TATGTT"
                     /label="-10 signal"
     -35_signal      complement(3903..3908)
                     /note="Plac promoter (drives lacZalpha;
                     counter-clockwise); TTTACA"
                     /label="-35 signal"
     protein_bind    complement(3928..3940)
                     /note="CAP protein binding site"
                     /bound_moiety="catabolite activator protein (CAP)"
                     /label="CAP protein binding site protein bind"
     rep_origin      complement(4232..4820)
                     /note="pMB1 origin of replication (counter-clockwise)
                     (RNAII -35 to RNA/DNA switch point)"
                     /label="rep origin"
     misc_RNA        complement(4232..4784)
                     /note="RNAII transcript (complementary strand)"
                     /label="RNA"
     -35_signal      4638..4643
                     /note="RNAI promoter (clockwise); TTGAAG"
                     /label="RNAI promoter (clockwise); TTGAAG -35 signal"
     -10_signal      4660..4665
                     /note="RNAI promoter (clockwise); GCTACA"
                     /label="RNAI promoter (clockwise); GCTACA -10 signal"
     misc_RNA        4674..4781
                     /note="RNAI transcript"
                     /label="RNAI transcript"
     -10_signal      complement(4794..4799)
                     /note="RNAII promoter (counter-clockwise); CGTAAT"
                     /label="-10 signal"
     -35_signal      complement(4815..4820)
                     /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 ctatgcggca tcagagcaga ttgtactgag
     1081 agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag
     1141 gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
     1201 gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc
     1261 agggttttcc cagtcacgac gttgtaaaac gacggccagt gaattcgagc tcggtacgcg
     1321 ccgggttttg gcgcctcccg cgggcgcccc cctcctcacg gcgagcgctg ccacgtcaga
     1381 cgaagggcgc agcgagcgtc ctgatccttc cgcccggacg ctcaggacag cggcccgctg
     1441 ctcataagac tcggccttag aaccccagta tcagcagaag gacattttag gacgggactt
     1501 gggtgactct agggcactgg ttttctttcc agagagcgga acaggcgagg aaaagtagtc
     1561 ccttctcggc gattctgcgg agggatctcc gtggggcggt gaacgccgat gattatataa
     1621 ggacgcgccg ggtgtggcac agctagttcc gtcgcagccg ggatttgggt cgcggttctt
     1681 gtttgtggat cgctgtgatc gtcacttggt gagtagcggg ctgctgggct ggccggggct
     1741 ttcgtggccg ccgggccgct cggtgggacg gaagcgtgtg gagagaccgc caagggctgt
     1801 agtctgggtc cgcgagcaag gttgccctga actgggggtt ggggggagcg cagcaaaatg
     1861 gcggctgttc ccgagtcttg aatggaagac gcttgtgagg cgggctgtga ggtcgttgaa
     1921 acaaggtggg gggcatggtg ggcggcaaga acccaaggtc ttgaggcctt cgctaatgcg
     1981 ggaaagctct tattcgggtg agatgggctg gggcaccatc tggggaccct gacgtgaagt
     2041 ttgtcactga ctggagaact cggtttgtcg tctgttgcgg gggcggcagt tatggcggtg
     2101 ccgttgggca gtgcacccgt acctttggga gcgcgcgccc tcgtcgtgtc gtgacgtcac
     2161 ccgttctgtt ggcttataat gcagggtggg gccacctgcc ggtaggtgtg cggtaggctt
     2221 ttctccgtcg caggacgcag ggttcgggcc tagggtaggc tctcctgaat cgacaggcgc
     2281 cggacctctg gtgaggggag ggataagtga ggcgtcagtt tctttggtcg gttttatgta
     2341 cctatcttct taagtagctg aagctccggt tttgaactat gcgctcgggg ttggcgagtg
     2401 tgttttgtga agttttttag gcaccttttg aaatgtaatc atttgggtca atatgtaatt
     2461 ttcagtgtta gactagtaaa ttgtccgcta aattctggcc gtttttggct tttttgttag
     2521 acctggaggt ccgccaccgc caccatggac cctaaaaaaa agagaaaggt ggacccaaag
     2581 aagaaacgca aagttgatcc gaagaagaaa aggaaagttg gctaccccta cgacgtgccc
     2641 gactacgccg gcagtggcgc cggcaccggc ggctccatgt caaagacgat cgtgctgtct
     2701 gtaggtgaag ccacacgcac cctcactgag atacagtcca cggccgatag gcagattttt
     2761 gaggagaagg tgggtccact ggtgggtagg cttcgactga cggcctccct gaggcaaaac
     2821 ggcgctaaaa cggcttatcg cgttaacctc aagctggacc aggccgatgt cgttgatagt
     2881 ggactgccca aagtcaggta cacacaggtg tggagccacg atgtgaccat cgtggctaac
     2941 tcaacagagg cgtcccggaa gtctctgtac gacctgacaa agtccctggt tgccacctct
     3001 caggtggagg atctggtggt taatctcgtg cccctcggac ggggttccac gagcgggtca
     3061 ggtggcggct ccatgtcaaa gacgatcgtg ctgtctgtag gtgaagccac acgcaccctc
     3121 actgagatac agtccacggc cgataggcag atttttgagg agaaggtggg tccactggtg
     3181 ggtaggcttc gactgacggc ctccctgagg caaaacggcg ctaaaacggc ttatcgcgtt
     3241 aacctcaagc tggaccaggc cgatgtcgtt gatagtggac tgcccaaagt caggtacaca
     3301 caggtgtgga gccacgatgt gaccatcgtg gctaactcaa cagaggcgtc ccggaagtct
     3361 ctgtacgacc tgacaaagtc cctggttgcc acctctcagg tggaggatct ggtggttaat
     3421 ctcgtgcccc tcggacgggg ttccggtgga tccggttccg gacgggctga cgcattggac
     3481 gattttgatc tggatatgct gggaagtgac gccctcgatg attttgacct tgacatgctt
     3541 ggttcggatg cccttgatga ctttgacctc gacatgctcg gcagtgacgc ccttgatgat
     3601 ttcgacctgg acatgctgat taactgaggt acctgaataa atgagggccc gtttaaaccc
     3661 gctgatcagt tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     3721 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     3781 gtatcttatc tgtttaaacc tgcaggcatg caagcttggc gtaatcatgg tcatagctgt
     3841 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     3901 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     3961 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     4021 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     4081 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     4141 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     4201 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     4261 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     4321 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     4381 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     4441 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     4501 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     4561 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     4621 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     4681 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     4741 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     4801 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     4861 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     4921 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     4981 ggtctgacag
//
