pLKO.1-hygro

价格:1250元

联系方式:I47-825O-882O

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载体基本信息

出品公司: Addgene
载体名称: pLKO.1-hygro
质粒类型: 慢病毒载体;RNAi载体
高拷贝/低拷贝: 低拷贝
克隆方法: AgeI,EcoRI
启动子: U6
载体大小: 7539 bp
5' 测序引物及序列: LKO.15' :GACTATCATATGCTTACCGT
3' 测序引物及序列: SV40pA-R : GAAATTTGTGATGCTATTGC
载体标签: --
载体抗性: 氨苄青霉素
筛选标记: hygromycin
克隆菌株: Stbl3
宿主细胞(系): 分哺乳动物细胞
备注:
3rd gen lentiviral backbone for cloning and expression of new shRNA sequences.
 
产品目录号: #24150
稳定性: 稳表达
组成型/诱导型: 组成型
病毒/非病毒: 慢病毒

载体质粒图谱和多克隆位点信息

pLKO.1-hygro载体图谱



载体简介

载体序列

LOCUS       Exported                7539 bp ds-DNA     circular SYN 08-AUG-2016
DEFINITION  synthetic circular DNA
KEYWORDS    pLKO.1 hygro
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7539)
  AUTHORS   Biofeng Lab.
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7539
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     protein_bind    107..128
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding site"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        143..173
                     /note="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="lac operator"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     205..221
                     /note="M13 rev"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        242..260
                     /note="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        288..514
                     /note="RSV promoter"
                     /note="Rous sarcoma virus enhancer/promoter"
     misc_feature    515..695
                     /note="truncHIV1 3'LTR"
     LTR             515..695
                     /note="5' LTR"
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    742..867
                     /note="HIV-1 Psi"
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1360..1593
                     /note="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     promoter        2120..2360
                     /note="U6 promoter"
                     /note="RNA polymerase III promoter for human U6 snRNA"
     misc_feature    2366..2395
                     /note="MCS"
     misc_feature    2431..2548
                     /note="cPPT/CTS"
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    2435..2450
                     /note="cPPT"
     promoter        2597..3107
                     /note="hPGK promoter"
                     /note="human phosphoglycerate kinase 1 promoter"
     CDS             3197..4222
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /note="HygR"
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     primer_bind     complement(4246..4262)
                     /note="KS primer"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    4328..4343
                     /note="cPPT"
     LTR             4345..4578
                     /note="3' LTR (Delta-U3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4650..4771
                     /note="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      4811..4946
                     /note="SV40 ori"
                     /note="SV40 origin of replication"
     promoter        complement(4967..4985)
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4995..5011)
                     /note="M13 fwd"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      5153..5608
                     /direction=RIGHT
                     /note="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        5634..5738
                     /gene="bla"
                     /note="bla"
                     /note="AmpR promoter"
     CDS             5739..6599
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      6770..7358
                     /direction=RIGHT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg
      241 caattaaccc tcactaaagg gaacaaaagc tggagctgca agcttaatgt agtcttatgc
      301 aatactcttg tagtcttgca acatggtaac gatgagttag caacatgcct tacaaggaga
      361 gaaaaagcac cgtgcatgcc gattggtgga agtaaggtgg tacgatcgtg ccttattagg
      421 aaggcaacag acgggtctga catggattgg acgaaccact gaattgccgc attgcagaga
      481 tattgtattt aagtgcctag ctcgatacat aaacgggtct ctctggttag accagatctg
      541 agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc
      601 ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct
      661 cagacccttt tagtcagtgt ggaaaatctc tagcagtggc gcccgaacag ggacttgaaa
      721 gcgaaaggga aaccagagga gctctctcga cgcaggactc ggcttgctga agcgcgcacg
      781 gcaagaggcg aggggcggcg actggtgagt acgccaaaaa ttttgactag cggaggctag
      841 aaggagagag atgggtgcga gagcgtcagt attaagcggg ggagaattag atcgcgatgg
      901 gaaaaaattc ggttaaggcc agggggaaag aaaaaatata aattaaaaca tatagtatgg
      961 gcaagcaggg agctagaacg attcgcagtt aatcctggcc tgttagaaac atcagaaggc
     1021 tgtagacaaa tactgggaca gctacaacca tcccttcaga caggatcaga agaacttaga
     1081 tcattatata atacagtagc aaccctctat tgtgtgcatc aaaggataga gataaaagac
     1141 accaaggaag ctttagacaa gatagaggaa gagcaaaaca aaagtaagac caccgcacag
     1201 caagcggccg ctgatcttca gacctggagg aggagatatg agggacaatt ggagaagtga
     1261 attatataaa tataaagtag taaaaattga accattagga gtagcaccca ccaaggcaaa
     1321 gagaagagtg gtgcagagag aaaaaagagc agtgggaata ggagctttgt tccttgggtt
     1381 cttgggagca gcaggaagca ctatgggcgc agcgtcaatg acgctgacgg tacaggccag
     1441 acaattattg tctggtatag tgcagcagca gaacaatttg ctgagggcta ttgaggcgca
     1501 acagcatctg ttgcaactca cagtctgggg catcaagcag ctccaggcaa gaatcctggc
     1561 tgtggaaaga tacctaaagg atcaacagct cctggggatt tggggttgct ctggaaaact
     1621 catttgcacc actgctgtgc cttggaatgc tagttggagt aataaatctc tggaacagat
     1681 ttggaatcac acgacctgga tggagtggga cagagaaatt aacaattaca caagcttaat
     1741 acactcctta attgaagaat cgcaaaacca gcaagaaaag aatgaacaag aattattgga
     1801 attagataaa tgggcaagtt tgtggaattg gtttaacata acaaattggc tgtggtatat
     1861 aaaattattc ataatgatag taggaggctt ggtaggttta agaatagttt ttgctgtact
     1921 ttctatagtg aatagagtta ggcagggata ttcaccatta tcgtttcaga cccacctccc
     1981 aaccccgagg ggacccgaca ggcccgaagg aatagaagaa gaaggtggag agagagacag
     2041 agacagatcc attcgattag tgaacggatc tcgacggtat cgatcacgag actagcctcg
     2101 agcggccgcc cccttcaccg agggcctatt tcccatgatt ccttcatatt tgcatatacg
     2161 atacaaggct gttagagaga taattggaat taatttgact gtaaacacaa agatattagt
     2221 acaaaatacg tgacgtagaa agtaataatt tcttgggtag tttgcagttt taaaattatg
     2281 ttttaaaatg gactatcata tgcttaccgt aacttgaaag tatttcgatt tcttggcttt
     2341 atatatcttg tggaaaggac gaaacaccgg tccgcaggta tgcacgcgtg aattctcgac
     2401 ctcgagacaa atggcagtat tcatccacaa ttttaaaaga aaagggggga ttggggggta
     2461 cagtgcaggg gaaagaatag tagacataat agcaacagac atacaaacta aagaattaca
     2521 aaaacaaatt acaaaaattc aaaattttcg ggtttattac agggacagca gagatccact
     2581 ttggccgcgg ctcgaggggg ttggggttgc gccttttcca aggcagccct gggtttgcgc
     2641 agggacgcgg ctgctctggg cgtggttccg ggaaacgcag cggcgccgac cctgggtctc
     2701 gcacattctt cacgtccgtt cgcagcgtca cccggatctt cgccgctacc cttgtgggcc
     2761 ccccggcgac gcttcctgct ccgcccctaa gtcgggaagg ttccttgcgg ttcgcggcgt
     2821 gccggacgtg acaaacggaa gccgcacgtc tcactagtac cctcgcagac ggacagcgcc
     2881 agggagcaat ggcagcgcgc cgaccgcgat gggctgtggc caatagcggc tgctcagcag
     2941 ggcgcgccga gagcagcggc cgggaagggg cggtgcggga ggcggggtgt ggggcggtag
     3001 tgtgggccct gttcctgccc gcgcggtgtt ccgcattctg caagcctccg gagcgcacgt
     3061 cggcagtcgg ctccctcgtt gaccgaatca ccgacctctc tccccagggg gatccaccgg
     3121 atcaagcttg ccagctgggg cgccctctgg taaggttggg aagccctgca gatctgatcc
     3181 cggggggcaa tgagatatga aaaagcctga actcaccgcg acgtctgtcg agaagtttct
     3241 gatcgaaaag ttcgacagcg tctccgacct gatgcagctc tcggagggcg aagaatctcg
     3301 tgctttcagc ttcgatgtag gagggcgtgg atatgtcctg cgggtaaata gctgcgccga
     3361 tggtttctac aaagatcgtt atgtttatcg gcactttgca tcggccgcgc tcccgattcc
     3421 ggaagtgctt gacattgggg aatttagcga gagcctgacc tattgcatct cccgccgtgc
     3481 acagggtgtc acgttgcaag acctgcctga aaccgaactg cccgctgttc tgcagccggt
     3541 cgcggaggcc atggatgcga tcgctgcggc cgatcttagc cagacgagcg ggttcggccc
     3601 attcggaccg caaggaatcg gtcaatacac tacatggcgt gatttcatat gcgcgattgc
     3661 tgatccccat gtgtatcact ggcaaactgt gatggacgac accgtcagtg cgtccgtcgc
     3721 gcaggctctc gatgagctga tgctttgggc cgaggactgc cccgaagtcc ggcacctcgt
     3781 gcacgcggat ttcggctcca acaatgtcct gacggacaat ggccgcataa cagcggtcat
     3841 tgactggagc gaggcgatgt tcggggattc ccaatacgag gtcgccaaca tcttcttctg
     3901 gaggccgtgg ttggcttgta tggagcagca gacgcgctac ttcgagcgga ggcatccgga
     3961 gcttgcagga tcgccgcggc tccgggcgta tatgctccgc attggtcttg accaactcta
     4021 tcagagcttg gttgacggca atttcgatga tgcagcttgg gcgcagggtc gatgcgacgc
     4081 aatcgtccga tccggagccg ggactgtcgg gcgtacacaa atcgcccgca gaagcgcggc
     4141 cgtctggacc gatggctgtg tagaagtact cgccgatagt ggaaaccgac gccccagcac
     4201 tcgtccgagg gcaaaggaat agagtagatg ccgaccggga tctatcgata ccgtcgacct
     4261 cgaggggggg cccggtacct ttaagaccaa tgacttacaa ggcagctgta gatcttagcc
     4321 actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga agacaagatc
     4381 tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg
     4441 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
     4501 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
     4561 tgtggaaaat ctctagcagt agtagttcat gtcatcttat tattcagtat ttataacttg
     4621 caaagaaatg aatatcagag agtgagagga acttgtttat tgcagcttat aatggttaca
     4681 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     4741 gtggtttgtc caaactcatc aatgtatctt atcatgtctg gctctagcta tcccgcccct
     4801 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     4861 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
     4921 gtagtgagga ggcttttttg gaggcctagg gacgtaccca attcgcccta tagtgagtcg
     4981 tattacgcgc gctcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt
     5041 acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag
     5101 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg ggacgcgccc
     5161 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     5221 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     5281 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     5341 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     5401 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     5461 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     5521 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     5581 tttaacaaaa tattaacgct tacaatttag gtggcacttt tcggggaaat gtgcgcggaa
     5641 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     5701 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     5761 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     5821 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     5881 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     5941 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     6001 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     6061 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     6121 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     6181 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     6241 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     6301 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     6361 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     6421 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     6481 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     6541 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     6601 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     6661 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     6721 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     6781 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     6841 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     6901 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     6961 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     7021 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     7081 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     7141 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     7201 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     7261 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     7321 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     7381 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     7441 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     7501 cgaccgagcg cagcgagtca gtgagcgagg aagcggaag
//

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