LOCUS       pUBC_HA_L7Ae~VP6        4583 bp    DNA     circular UNA 07-JUL-2015
DEFINITION  Transient expression of humanized A fulgidus L7Ae, fused to the
            VP64 transcription activator, in human cells, under a UBC promoter.
ACCESSION   urn.local...1435874685495.40
VERSION     urn.local...1435874685495.40
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,<3383..>3823,<3824..>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    2540..2549
                     /created_by="User"
                     /label="Kozak_Sequence"
     CDS             2546..3040
                     /created_by="User"
                     /modified_by="User"
                     /label="3xNLS_HA_L7Ae (A. Fugildis)"
     sig_peptide     2549..2572
                     /created_by="User"
                     /label="NLS"
     sig_peptide     2573..2596
                     /created_by="User"
                     /label="NLS"
     sig_peptide     2597..2620
                     /created_by="User"
                     /label="NLS"
     misc_feature    2624..2650
                     /created_by="User"
                     /label="HA_Tag"
     CDS             3041..3220
                     /created_by="User"
                     /label="VP64_Transcription_Activator"
     terminator      3227..3229
                     /created_by="User"
                     /label="stop"
     terminator      3231..3233
                     /created_by="User"
                     /label="stop"
     terminator      3235..3237
                     /created_by="User"
                     /label="stop"
     terminator      3263..3382
                     /created_by="User"
                     /label="SV40_pA_terminator"
     misc_feature    complement(3297..3316)
                     /created_by="User"
                     /label="SV40pA_Primer_from_Genewiz"
                     /note="Geneious type: primer_bind_reverse"
     primer_bind     3351..3370
                     /created_by="User"
                     /label="EBV_Rev_Primer"
     misc_feature    complement(3423..3439)
                     /created_by="User"
                     /label="M13R Primer from GeneWiz"
                     /note="Geneious type: primer_bind_reverse"
     -10_signal      complement(3472..3477)
                     /note="Plac promoter (drives lacZalpha;
                     counter-clockwise); TATGTT"
                     /label="-10 signal"
     -35_signal      complement(3496..3501)
                     /note="Plac promoter (drives lacZalpha;
                     counter-clockwise); TTTACA"
                     /label="-35 signal"
     protein_bind    complement(3521..3533)
                     /note="CAP protein binding site"
                     /bound_moiety="catabolite activator protein (CAP)"
                     /label="CAP protein binding site protein bind"
     rep_origin      complement(3825..4413)
                     /note="pMB1 origin of replication (counter-clockwise)
                     (RNAII -35 to RNA/DNA switch point)"
                     /label="rep origin"
     misc_RNA        complement(3825..4377)
                     /note="RNAII transcript (complementary strand)"
                     /label="RNA"
     -35_signal      4231..4236
                     /note="RNAI promoter (clockwise); TTGAAG"
                     /label="RNAI promoter (clockwise); TTGAAG -35 signal"
     -10_signal      4253..4258
                     /note="RNAI promoter (clockwise); GCTACA"
                     /label="RNAI promoter (clockwise); GCTACA -10 signal"
     misc_RNA        4267..4374
                     /note="RNAI transcript"
                     /label="RNAI transcript"
     -10_signal      complement(4387..4392)
                     /note="RNAII promoter (counter-clockwise); CGTAAT"
                     /label="-10 signal"
     -35_signal      complement(4408..4413)
                     /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 acggccaccg ccaccatgga ccctaaaaaa aagagaaagg tggacccaaa
     2581 gaagaaacgc aaagttgatc cgaagaagaa aaggaaagtt ggctacccct acgacgtgcc
     2641 cgactacgcc ggaagtggtg ccggcaccgg cggcatgtac gtgcgcttcg aggtgcccga
     2701 ggacatgcag aacgaggccc tgagcctgct ggaaaaagtg cgcgagagcg gcaaagtgaa
     2761 gaagggcacc aacgaaacca ccaaggccgt ggaacggggc ctggccaagc tggtgtatat
     2821 cgccgaggac gtggaccccc ccgagattgt ggcccatctg cccctgctgt gcgaagagaa
     2881 gaacgtgccc tacatctacg tgaagtccaa gaacgacctg ggcagagccg tgggcatcga
     2941 ggtgccatgt gcctctgccg ccatcatcaa cgagggcgag ctgcggaaag aactgggcag
     3001 cctggtggaa aagatcaagg gcctgcagaa gggttccggt ggatccggtt ccggacgggc
     3061 tgacgcattg gacgattttg atctggatat gctgggaagt gacgccctcg atgattttga
     3121 ccttgacatg cttggttcgg atgcccttga tgactttgac ctcgacatgc tcggcagtga
     3181 cgcccttgat gatttcgacc tggacatgct gattaactga ggtacctgaa taaatgaggg
     3241 cccgtttaaa cccgctgatc agttgtttat tgcagcttat aatggttaca aataaagcaa
     3301 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     3361 caaactcatc aatgtatctt atctgtttaa acctgcaggc atgcaagctt ggcgtaatca
     3421 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     3481 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     3541 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     3601 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     3661 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     3721 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     3781 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     3841 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     3901 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     3961 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     4021 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     4081 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     4141 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     4201 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     4261 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     4321 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     4381 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     4441 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     4501 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     4561 tatgagtaaa cttggtctga cag
//
