| 質粒名稱: | pSL21-VEX |
|---|---|
| 出品公司: | Addgene |
| 目錄編號: | 158230 |
| 存儲實驗室: | Junwei Shi |
|
載體骨架基本信息 |
|
| 載體名稱: | pSL21-VEX |
| 載體出品公司: | Clontech |
| 空載體大小: | 6619bp |
| 完整質粒大小: | 6619bp |
| 載體類型: | 哺乳細胞表達, Retroviral, CRISPR |
| 載體抗性: | 氨芐霉素 |
| 生長溫度: | 37 ℃ |
| 宿主菌: | NEB Stable |
| 拷貝數: | 高拷貝 |
|
插入基因信息 |
|
| 啟動子: | U6 promoter for crRNA expression and EFS promoterfor VEX expression |
| 克隆方法: | 限制性內切酶 |
|
文獻引用方法 |
|
| 材料和方法部分: | pSL21-VEX was a gift from Junwei Shi (Addgene plasmid # 158230 ; //n2t.net/addgene:158230 ; RRID:Addgene_158230) |
| 參考文獻部分: | In vivo CD8(+) T cell CRISPR screening reveals control by Fli1 in infection and cancer. Chen Z, Arai E, Khan O, Zhang Z, Ngiow SF, He Y, Huang H, Manne S, Cao Z, Baxter AE, Cai Z, Freilich E, Ali MA, Giles JR, Wu JE, Greenplate AR, Hakeem MA, Chen Q, Kurachi M, Nzingha K, Ekshyyan V, Mathew D, Wen Z, Speck NA, Battle A, Berger SL, Wherry EJ, Shi J. Cell. 2021 Mar 4;184(5):1262-1280.e22. doi: 10.1016/j.cell.2021.02.019. Epub 2021 Feb 25. 10.1016/j.cell.2021.02.019 PubMed 33636129 |
買家導航
pSL21-VEX載體質粒基本信息
pSL21-VEX質粒圖譜載體圖譜和pSL21-VEX載體序列質粒序列多克隆位點信息
pSL21-VEX質粒載體簡介
pSL21-VEX質粒序列載體序列
LOCUS pSL21-VEX 6619 bp DNA circular SYN 14-APR-2025
DEFINITION Retroviral expression plasmid of sgRNA with violet-excited
fluorescent protein (VEX).
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6619)
AUTHORS Chen Z, Arai E, Khan O, Zhang Z, Ngiow SF, He Y, Huang H, Manne S,
Cao Z, Baxter AE, Cai Z, Freilich E, Ali MA, Giles JR, Wu JE,
Greenplate AR, Hakeem MA, Chen Q, Kurachi M, Nzingha K, Ekshyyan V,
Mathew D, Wen Z, Speck NA, Battle A, Berger SL, Wherry EJ, Shi J
TITLE In vivo CD8(+) T cell CRISPR screening reveals control by Fli1 in
infection and cancer.
JOURNAL Cell. 2021 Mar 4;184(5):1262-1280.e22. doi:
10.1016/j.cell.2021.02.019. Epub 2021 Feb 25.
PUBMED 33636129
REFERENCE 2 (bases 1 to 6619)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Apr 14, 2025 from SnapGene Server 8.0.1
//gt12365.cn
COMMENT SGRef: number: 1; type: "Journal Article"; doi:
"10.1016/j.cell.2021.02.019"; journalName: "Cell"; date: "2021-03-4-
4"; volume: "184"; issue: "5"; pages: "1262-1280.e22"
COMMENT Sequence Label: pSL21-VEX
FEATURES Location/Qualifiers
source 1..6619
/mol_type="other DNA"
/organism="synthetic DNA construct"
LTR 1..517
/label=5' LTR
/note="5' long terminal repeat from murine embryonic stem
cell virus"
misc_feature 581..922
/label=MESV Psi
/note="packaging signal of murine embryonic stem cell
virus"
CDS 989..1405
/label=gag (truncated)
/note="truncated Moloney murine leukemia virus (MMLV) gag
gene lacking the start codon"
primer_bind 1335..1357
/label=pLXSN 5'
/note="Murine stem cell virus, forward primer. Also called
MSCV"
primer_bind 1373..1389
/label=pBABE 5'
/note="Psi packaging signal, forward primer for pBABE
vectors"
promoter 1408..1656
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA
(Domitrovich & Kunkel, 2003)"
primer_bind 1408..1428
/label=hU6-F
/note="Human U6 promoter, forward primer"
primer_bind 1579..1598
/label=LKO.1 5'
/note="Human U6 promoter, forward primer"
misc_feature 1820..1937
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 2006..2217
/label=EF-1-alpha core promoter
/note="core promoter for human elongation factor
EF-1-alpha"
CDS join(2246..2248,2249..2251,2252..2962)
/codon_start=1
/product="the original enhanced GFP (Yang et al., 1996)"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLFIQSALSKDPNEKRDHMVLL
EFVTAAGITHGMDELYK"
primer_bind complement(2291..2312)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(2552..2571)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 2899..2920
/label=EGFP-C
/note="EGFP, forward primer"
LTR 3014..3528
/label=3' LTR
/note="3' long terminal repeat from murine embryonic stem
cell virus"
protein_bind 3690..3706
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/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)."
promoter complement(join(3714..3720,3721..3738,3739..3744))
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3759..3780
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(3897..3914)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4068..4656)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4148..4167)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(join(4827..5618,5619..5687))
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 5450..5469
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5688..5792)
/gene="bla"
/label=AmpR promoter
primer_bind 5860..5878
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
primer_bind complement(5916..5938)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 6038..6057
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 6251..6273
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 6401..6420
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
ORIGIN
1 aatgaaagac cccacctgta ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc
61 atggaaaata cataactgag aatagagaag ttcagatcaa ggttaggaac agagagacag
121 cagaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
181 gaacagatgg tccccagatg cggtcccgcc ctcagcagtt tctagagaac catcagatgt
241 ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt
301 tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac
361 ccctcactcg gcgcgccagt cctccgatag actgcgtcgc ccgggtaccc gtattcccaa
421 taaagcctct tgctgtttgc atccgaatcg tggactcgct gatccttggg agggtctcct
481 cagattgatt gactgcccac ctcgggggtc tttcatttgg aggttccacc gagatttgga
541 gacccctgcc cagggaccac cgaccccccc gccgggaggt aagctggcca gcggtcgttt
601 cgtgtctgtc tctgtctttg tgcgtgtttg tgccggcatc taatgtttgc gcctgcgtct
661 gtactagtta gctaactagc tctgtatctg gcggacccgt ggtggaactg acgagttctg
721 aacacccggc cgcaaccctg ggagacgtcc cagggacttt gggggccgtt tttgtggccc
781 gacctgagga agggagtcga tgtggaatcc gaccccgtca ggatatgtgg ttctggtagg
841 agacgagaac ctaaaacagt tcccgcctcc gtctgaattt ttgctttcgg tttggaaccg
901 aagccgcgcg tcttgtctgc tgcagcgctg cagcatcgtt ctgtgttgtc tctgtctgac
961 tgtgtttctg tatttgtctg aaaattaggg ccagactgtt accactccct taagtttgac
1021 cttaggtcac tggaaagatg tcgagcggat cgctcacaac cagtcggtag atgtcaagaa
1081 gagacgttgg gttaccttct gctctgcaga atggccaacc tttaacgtcg gatggccgcg
1141 agacggcacc tttaaccgag acctcatcac ccaggttaag atcaaggtct tttcacctgg
1201 cccgcatgga cacccagacc aggtccccta catcgtgacc tgggaagcct tggcttttga
1261 cccccctccc tgggtcaagc cctttgtaca ccctaagcct ccgcctcctc ttcctccatc
1321 cgccccgtct ctcccccttg aacctcctcg ttcgaccccg cctcgatcct ccctttatcc
1381 agccctcact ccttctctag gcgccgggag ggcctatttc ccatgattcc ttcatatttg
1441 catatacgat acaaggctgt tagagagata attagaatta atttgactgt aaacacaaag
1501 atattagtac aaaatacgtg acgtagaaag taataatttc ttgggtagtt tgcagtttta
1561 aaattatgtt ttaaaatgga ctatcatatg cttaccgtaa cttgaaagta tttcgatttc
1621 ttggctttat atatcttgtg gaaaggacga aacaccgggg tcttcgagaa gacctgttta
1681 agagctatgc tggaaacagc atagcaagtt taaataaggc tagtccgtta tcaactttgc
1741 tggaaacagc aaagtggcac cgagtcggtg ctttttttga attctagatc ttgagacaaa
1801 tggcagtatt catccacaat tttaaaagaa aaggggggat tggggggtac agtgcagggg
1861 aaagaatagt agacataata gcaacagaca tacaaactaa agaattacaa aaacaaatta
1921 caaaaattca aaattttcgg gtttattaca gggacagcag agatccactt tggcgccggc
1981 tcgagtggct ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag
2041 ttggggggag gggtcggcaa ttgaaccggt gcctagagaa ggtggcgcgg ggtaaactgg
2101 gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata
2161 agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacaggtg
2221 tcgtgacgcg ggatccgcca ccatgatggt gagcaagggc gaggagctgt tcaccggggt
2281 ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg
2341 cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg
2401 caagctgccc gtgccctggc ccaccctcgt gaccaccttc agctacggcg tgcagtgctt
2461 cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg
2521 ctacgtccag gagcgcacca tcagcttcaa ggacgacggc aactacaaga cccgcgccga
2581 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa
2641 ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta
2701 tatcacggcc gacaagcaga agaacggcat caaggcgaac ttcaagatcc gccacaacat
2761 cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg
2821 ccccgtgctg ctgcccgaca accactacct gttcatccag tccgccctga gcaaagaccc
2881 caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactca
2941 cggcatggac gagctgtaca agagcttatc gataaaataa aagattttat ttagtctcca
3001 gaaaaagggg ggaatgaaag accccacctg taggtttggc aagctagctt aagtaacgcc
3061 attttgcaag gcatggaaaa tacataactg agaatagaga agttcagatc aaggttagga
3121 acagagagac agcagaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg
3181 gctcagggcc aagaacagat ggtccccaga tgcggtcccg ccctcagcag tttctagaga
3241 accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc ttatttgaac
3301 taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa
3361 agagcccaca acccctcact cggcgcgcca gtcctccgat agactgcgtc gcccgggtac
3421 ccgtgtatcc aataaaccct cttgcagttg catccgactt gtggtctcgc tgttccttgg
3481 gagggtctcc tctgagtgat tgactacccg tcagcggggg tctttcatgg gtaacagttt
3541 cttgaagttg gagaacaaca ttctgagggt aggagtcgaa tattaagtaa tcctgactca
3601 attagccact gttttgaatc cacatactcc aatactcctg aaatagttca ttatggacag
3661 cgcagaaaga gctggggaga attgtgaaat tgttatccgc tcacaattcc acacaacata
3721 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
3781 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
3841 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
3901 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
3961 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
4021 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
4081 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
4141 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
4201 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
4261 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
4321 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
4381 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
4441 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
4501 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
4561 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
4621 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
4681 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
4741 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
4801 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
4861 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
4921 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
4981 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
5041 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
5101 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
5161 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
5221 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
5281 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
5341 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
5401 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
5461 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
5521 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
5581 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
5641 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
5701 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
5761 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
5821 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
5881 tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
5941 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
6001 gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg
6061 agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc
6121 aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
6181 tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg
6241 ccagggtttt cccagtcacg acgttgtaaa acgacggcgc aaggaatggt gcatgcaagg
6301 agatggcgcc caacagtccc ccggccacgg ggcctgccac catacccacg ccgaaacaag
6361 cgctcatgag cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg gcgatatagg
6421 cgccagcaac cgcacctgtg gcgccggtga tgccggccac gatgcgtccg gcgtagaggc
6481 gattagtcca atttgttaaa gacaggatat cagtggtcca ggctctagtt ttgactcaac
6541 aatatcacca gctgaagcct atagagtacg agccatagat aaaataaaag attttattta
6601 gtctccagaa aaagggggg
//
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