pLKO.1-Hygro

 基本信息

启动子:U6

复制子:pUC

终止子:SV40 poly(A) signal

质粒分类:病毒系列，慢病毒干扰载体

质粒大小:7539bp

原核抗性:Amp

筛选标记:Hyg

克隆菌株:Stbl3

培养条件:37度

表达宿主:哺乳细胞

诱导方式:无须诱导，瞬时表达

5'测序引物:根据序列设计引物

3'测序引物:Sv40-polyA-R：GAAATTTGTGATGCTATTGC

质粒属性

载体宿主：哺乳细胞,慢病毒

载体用途：干扰沉默

基因种属：空载体

基因类型：shRNA

原核抗性：Amp

真核抗性：Hyg

质粒简介

pLKO.1 hygro is a replication-incompetent lentiviral vector for expression of shRNAs. pLKO.1 hygro can be introduced into cells via direct transfection, or can be converted into lentiviral particles for subsequent infection of a target cell line. Once introduced, the hygromycin resistance marker encoded in pLKO.1 allows for convenient stable selection.

质粒序列

LOCUS       Exported                7539 bp ds-DNA     circular SYN 09-SEP-2016

DEFINITION  synthetic circular DNA

ACCESSION   .

VERSION     .

KEYWORDS    pLKO.1-hygro

SOURCE      synthetic DNA construct

  ORGANISM  synthetic DNA construct

REFERENCE   1  (bases 1 to 7539)

  AUTHORS   xyzhang1971

  TITLE     Direct Submission

  JOURNAL   Exported Friday, September 9, 2016 from SnapGene Viewer 3.1.4

            http://www.miaolingbio.com

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        286..514

                     /note="RSV promoter"

                     /note="Rous sarcoma virus enhancer/promoter"

     LTR             515..695

                     /note="5' LTR (truncated)"

                     /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

                     /label=U6 Promoter

                     /note="U6 promoter"

                     /note="RNA polymerase III promoter for human U6 snRNA"

     misc_feature    2431..2548

                     /note="cPPT/CTS"

                     /note="central polypurine tract and central termination 

                     sequence of HIV-1"

     promoter        2597..3107

                     /note="hPGK promoter"

                     /note="human phosphoglycerate kinase 1 promoter"

     misc_feature    3122..4272

                     /note="Different region against pLKO.1-puro"

     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"

     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="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

//