pSingle-tTS-shRNA

LOCUS       pSingle-tTs-shRNA                6919 bp ds-DNA    circular SYN 11-1-2013
DEFINITION  Vector for doxycycline-inducible expression of an shRNA.
ACCESSION   .
VERSION     .
KEYWORDS    pSingle-tTs-shRNA
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6919)
  AUTHORS   Clontech
  TITLE     Direct Submission
  JOURNAL   Exported 2017/2/28 from SnapGene 2.3.2
            http://www.snapgene.com
COMMENT     Clone an annealed double-stranded oligonucleotide encoding the shRNA
            into the vector digested with XhoI and HindIII.
FEATURES             Location/Qualifiers
     source          1..6919
                     /organism="synthetic DNA construct"
                     /lab_host="Mammalian Cells"
                     /mol_type="other DNA"
     promoter        58..254
                     /note="SV40 promoter"
                     /note="SV40 early promoter"
     rep_origin      105..240
                     /note="SV40 ori"
                     /note="SV40 origin of replication"
     CDS             289..1083
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /note="NeoR/KanR"
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin)"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    1315..1363
                     /note="HSV TK poly(A) signal"
                     /note="herpesvirus thymidine kinase polyadenylation signal"
     enhancer        1449..1828
                     /note="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1829..2032
                     /note="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     CDS             2135..2983
                     /codon_start=1
                     /product="fusion of a hybrid tetracycline repressor TetR 
                     with a nuclear localization signal and a KRAB silencing 
                     domain"
                     /note="tTS"
                     /note="silences Tet-responsive promoters in the absence but
                     not the presence of doxycycline"
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     RNKQTLMNMLSEAILAKHHTRSAPLPTESWQQFLQENALSFRKALLVHRDGARLHIGTS
                     PTPPQFEQAEAQLRCLCDAGFSVEEALFILQSISHFTLGAVLEEQATNQIENNHVIDAA
                     PPLLQEAFNIQARTSAEMAFHFGLKSLIFGFSAQLDEKKHTPIEDGNKPKKKRKLAVSV
                     TFEDVAVLFTRDEWKKLDLSQRSLYREVMLENYSNLASMAGFLFTKPKVISLLQQGEDP
                     W"
     intron          3004..3576
                     /note="beta-globin intron"
                     /note="intron from rabbit beta-globin gene"
     polyA_site      3773..3828
                     /note="beta-globin poly(A) signal"
                     /note="rabbit beta-globin polyadenylation signal"
     promoter        4246..4577
                     /note="TREmod/U6 promoter"
                     /note="modified Tet-responsive promoter, consisting of 
                     seven repeats of the tet operator fused to a portion of the
                     human U6 promoter"
     protein_bind    4254..4272
                     /gene="tetO"
                     /bound_moiety="tetracycline repressor TetR"
                     /note="tet operator"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4290..4308
                     /gene="tetO"
                     /bound_moiety="tetracycline repressor TetR"
                     /note="tet operator"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4325..4343
                     /gene="tetO"
                     /bound_moiety="tetracycline repressor TetR"
                     /note="tet operator"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4361..4379
                     /gene="tetO"
                     /bound_moiety="tetracycline repressor TetR"
                     /note="tet operator"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4397..4415
                     /gene="tetO"
                     /bound_moiety="tetracycline repressor TetR"
                     /note="tet operator"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4432..4450
                     /gene="tetO"
                     /bound_moiety="tetracycline repressor TetR"
                     /note="tet operator"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4468..4486
                     /gene="tetO"
                     /bound_moiety="tetracycline repressor TetR"
                     /note="tet operator"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     rep_origin      complement(5100..5688)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5859..6719)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6720..6824)
                     /gene="bla"
                     /note="AmpR promoter"
ORIGIN
        1 ctcgattagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
       61 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
      121 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
      181 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
      241 taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
      301 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
      361 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
      421 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca
      481 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
      541 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
      601 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
      661 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
      721 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
      781 ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc
      841 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
      901 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
      961 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     1021 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     1081 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
     1141 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     1201 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg
     1261 ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa
     1321 taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt ttcgaggagc ttggcccatt
     1381 gcatacgttg tatccatatc ataatatgta catttatatt ggctcatgtc caacattacc
     1441 gccatgttga cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt
     1501 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
     1561 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
     1621 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
     1681 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
     1741 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
     1801 ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg
     1861 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag
     1921 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt
     1981 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctcgtttagt
     2041 gaaccgtcag atcgcctgga gacgccatcc acgctgtttt gacctccata gaagacaccg
     2101 ggaccgatcc agcctccgcg gccccgaatt caccatgtct agattagata aaagtaaagt
     2161 gattaacagc gcattagagc tgcttaatga ggtcggaatc gaaggtttaa caacccgtaa
     2221 actcgcccag aagctaggtg tagagcagcc tacattgtat tggcacgtgc gcaacaagca
     2281 gactcttatg aacatgcttt cagaggcaat actggcgaag catcacaccc gttcagcacc
     2341 gttaccgact gagagttggc agcagtttct ccaggaaaat gctctgagtt tccgtaaagc
     2401 attactggtc catcgtgatg gagcccgatt gcatataggg acctctccta cgccccccca
     2461 gtttgaacaa gcagaggcgc aactacgctg tctatgcgat gcagggtttt cggtcgagga
     2521 ggctcttttc attctgcaat ctatcagcca ttttacgttg ggtgcagtat tagaggagca
     2581 agcaacaaac cagatagaaa ataatcatgt gatagacgct gcaccaccat tattacaaga
     2641 ggcatttaat attcaggcga gaacctctgc tgaaatggcc ttccatttcg ggctgaaatc
     2701 attaatattt ggattttctg cacagttaga tgaaaaaaag catacaccca ttgaggatgg
     2761 taataaacca aaaaagaaga gaaagctagc agtgtcagtg acatttgaag atgtggctgt
     2821 gctctttact cgggacgagt ggaagaagct ggatctgtct cagagaagcc tgtaccgtga
     2881 ggtgatgctg gagaattaca gcaacctggc ctccatggca ggattcctgt ttaccaaacc
     2941 aaaggtgatc tccctgttgc agcaaggaga ggatccctgg taaatcgatc tgagaacttc
     3001 agggtgagtt tggggaccct tgattgttct ttctttttcg ctattgtaaa attcatgtta
     3061 tatggagggg gcaaagtttt cagggtgttg tttagaatgg gaagatgtcc cttgtatcac
     3121 catggaccct catgataatt ttgtttcttt cactttctac tctgttgaca accattgtct
     3181 cctcttattt tcttttcatt ttctgtaact ttttcgttaa actttagctt gcatttgtaa
     3241 cgaattttta aattcacttt tgtttatttg tcagattgta agtactttct ctaatcactt
     3301 ttttttcaag gcaatcaggg tatattatat tgtacttcag cacagtttta gagaacaatt
     3361 gttataatta aatgataagg tagaatattt ctgcatataa attctggctg gcgtggaaat
     3421 attcttattg gtagaaacaa ctacaccctg gtcatcatcc tgcctttctc tttatggtta
     3481 caatgatata cactgtttga gatgaggata aaatactctg agtccaaacc gggcccctct
     3541 gctaaccatg ttcatgcctt cttctctttc ctacagctcc tgggcaacgt gctggttgtt
     3601 gtgctgtctc atcattttgg caaagaattc actcctcagg tgcaggctgc ctatcagaag
     3661 gtggtggctg gtgtggccaa tgccctggct cacaaatacc actgagatct ttttccctct
     3721 gccaaaaatt atggggacat catgaagccc cttgagcatc tgacttctgg ctaataaagg
     3781 aaatttattt tcattgcaat agtgtgttgg aattttttgt gtctctcact cggaaggaca
     3841 tatgggaggg caaatcattt aaaacatcag aatgagtatt tggtttagag tttggcaaca
     3901 tatgcccata tgctggctgc catgaacaaa ggttggctat aaagaggtca tcagtatatg
     3961 aaacagcccc ctgctgtcca ttccttattc catagaaaag ccttgacttg aggttagatt
     4021 ttttttatat tttgttttgt gttatttttt tctttaacat ccctaaaatt ttccttacat
     4081 gttttactag ccagattttt cctcctctcc tgactactcc cagtcatagc tgtccctctt
     4141 ctcttatgaa ctcgactgaa gctcacattt ccccgaaaag tgccacctga cgtaactata
     4201 acggtcctaa ggtagcgaaa gctcagatct ctttcgtctt cacttgagtt tactccctat
     4261 cagtgataga gaacgtatgt cgagtttact ccctatcagt gatagagaac gatgtcgagt
     4321 ttactcccta tcagtgatag agaacgtatg tcgagtttac tccctatcag tgatagagaa
     4381 cgtatgtcga gtttactccc tatcagtgat agagaacgta tgtcgagttt atccctatca
     4441 gtgatagaga acgtatgtcg agtttactcc ctatcagtga tagagaacgt atgtcgaggt
     4501 aggcgtgtac ggtcatatgc ttaccgtaac ttgaaagtat ttcgatttct tggctttata
     4561 tatcttgtgg aaaggacgac tcgagggaca agcttatcta tgtcgggtgc ggatcttgtg
     4621 gaaaggacga ctcgagggac aagcttatct atgtcgggtg cggagaaaga ggtaatgaaa
     4681 tggcattatg ggtattatgg gtctgcatta atgagcttgg cgagattttc aggagctaag
     4741 gaagctaaaa tggagaaaaa aatcactgga tataccaccg ttgatatatc ccaatggcat
     4801 cgtaaagaac attttgaggc atttcagtca gttgctcaat gtacctataa ccagaccgtt
     4861 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     4921 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     4981 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     5041 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     5101 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     5161 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     5221 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     5281 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     5341 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     5401 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     5461 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     5521 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     5581 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     5641 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     5701 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     5761 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     5821 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     5881 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     5941 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     6001 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     6061 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     6121 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     6181 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     6241 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     6301 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     6361 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     6421 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaacacgg
     6481 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     6541 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     6601 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     6661 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     6721 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     6781 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     6841 gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt
     6901 atcacgaggc cctttcgtc
//