基本信息
啟動子:U6
復(fù)制子:pUC
終止子:SV40 poly(A) signal
質(zhì)粒分類:病毒系列，慢病毒干擾載體
質(zhì)粒大小:7539bp
原核抗性:Amp
篩選標(biāo)記:Hyg
克隆菌株:Stbl3
培養(yǎng)條件:37度
表達(dá)宿主:哺乳細(xì)胞
誘導(dǎo)方式:無須誘導(dǎo)，瞬時表達(dá)
5'測序引物:根據(jù)序列設(shè)計引物
3'測序引物:Sv40-polyA-R：GAAATTTGTGATGCTATTGC
質(zhì)粒屬性
載體宿主：哺乳細(xì)胞,慢病毒
載體用途：干擾沉默
基因種屬：空載體
基因類型：shRNA
原核抗性：Amp
真核抗性：Hyg
質(zhì)粒簡介
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.
質(zhì)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
//
