| 质粒名称: | pLenti SpBsmBI sgRNA Puro |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 62207 |
| 存储实验室: | Rene Maehr |
|
载体骨架基本信息 |
|
| 载体名称: | pLV |
| 载体出品公司: | Addgene plasmid number: 36297 |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | 新霉素（G418） |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | -- |
| 物种来源: | -- |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | AgeI(未破坏) |
| 3' 克隆位点: | NotI (未破坏) |
| 5' 测序引物 | CMV-F |
| 3' 测序引物 | SV40pA-R |
|
文献引用方法 |
|
| 材料和方法部分: | pLenti SpBsmBI sgRNA Puro was a gift from Rene Maehr (Addgene plasmid # 62207 ; http://n2t.net/addgene:62207 ; RRID:Addgene_62207) |
| 参考文献部分: | Transcriptional Regulation with CRISPR/Cas9 Effectors in Mammalian Cells. Pham H, Kearns NA, Maehr R. Methods Mol Biol. 2016;1358:43-57. doi: 10.1007/978-1-4939-3067-8_3. 10.1007/978-1-4939-3067-8_3 PubMed 26463376 |
买家导航
载体基本信息
载体图谱质粒图谱和多克隆位点信息
载体简介
载体序列
LOCUS pLenti SpBsmBI sgRNA Puro 8475 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION An empty sgRNA expression vector for expression of sgRNA for Sp
Cas9.
ACCESSION .
VERSION .
KEYWORDS pLenti SpBsmBI sgRNA Puro
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8475)
AUTHORS Pham H, Kearns NA, Maehr R
TITLE Transcriptional Regulation with CRISPR/Cas9 Effectors in Mammalian
Cells.
JOURNAL Methods Mol Biol. 2016;1358:43-57. doi: 10.1007/978-1-4939-3067-8_3.
PUBMED 26463376
REFERENCE 2 (bases 1 to 8475)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..8475
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(20..40)
/label=T3
/note="T3 promoter, forward primer"
promoter complement(20..38)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(59..75)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(59..75)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(72..94)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 83..99
/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(107..137)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 152..173
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(290..307)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(461..1049)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(541..560)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(1220..2080)
/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 1843..1862
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(2081..2185)
/gene="bla"
/label=AmpR promoter
rep_origin complement(2211..2666)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2348..2369)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 2560..2579
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2793..2815
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 2807..2824
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 2808..2824
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 2834..2853
/label=T7
/note="T7 promoter, forward primer"
promoter 2834..2852
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
rep_origin 2873..3008
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind complement(2927..2946)
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
polyA_signal 3048..3169
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3059..3078)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 3113..3132
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
LTR 3241..3474
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
misc_feature 3561..4149
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
CDS 3667..3678
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
primer_bind 4076..4096
/label=WPRE-R
/note="WPRE, reverse primer"
promoter complement(4778..5959)
/label=EF-1-alpha promoter
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
intron 4787..5729
/label=EF-1-alpha intron A
/note="intron upstream of the start codon of human
EF-1-alpha"
primer_bind complement(4810..4830)
/label=EF1a-F
/note="Human elongation factor-1a promoter, forward primer"
misc_feature 6164..6280
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 6311..6551
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
primer_bind 6311..6331
/label=hU6-F
/note="Human U6 promoter, forward primer"
primer_bind 6482..6501
/label=LKO.1 5'
/note="Human U6 promoter, forward primer"
misc_RNA 6579..6654
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
misc_feature 7162..7395
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
misc_feature 7888..8013
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
LTR 8060..8240
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
promoter complement(8241..8467)
/label=RSV promoter
/note="Rous sarcoma virus enhancer/promoter"
ORIGIN
1 gcagctccag cttttgttcc ctttagtgag ggttaattgc gcgcttggcg taatcatggt
61 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
121 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
181 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
241 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
301 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
361 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
421 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
481 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
541 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
601 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
661 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
721 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
781 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
841 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
901 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
961 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
1021 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
1081 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
1141 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
1201 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
1261 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
1321 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
1381 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
1441 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
1501 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
1561 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
1621 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
1681 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
1741 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
1801 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
1861 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
1921 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
1981 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
2041 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
2101 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
2161 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct aaattgtaag
2221 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca
2281 ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag
2341 tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg
2401 gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt
2461 tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag
2521 agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc
2581 gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc
2641 gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc aggctgcgca actgttggga
2701 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc
2761 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc
2821 cagtgagcgc gcgtaatacg actcactata gggcgaattg ggtacgtccc taggcctcca
2881 aaaaagcctc ctcactactt ctggaatagc tcagaggccg aggcggcctc ggcctctgca
2941 taaataaaaa aaattagtca gccatggggc ggagaatggg cggaactggg cggagttagg
3001 ggcgggatgg gcggagttag gggcgggata gctagagcca gacatgataa gatacattga
3061 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
3121 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagttc ctctcactct
3181 ctgatattca tttctttgca agttataaat actgaataat aagatgacat gaactactac
3241 tgctagagat tttccacact gactaaaagg gtctgaggga tctctagtta ccagagtcac
3301 acaacagacg ggcacacact acttgaagca ctcaaggcaa gctttattga ggcttaagca
3361 gtgggttccc tagttagcca gagagctccc aggctcagat ctggtctaac cagagagacc
3421 cagtacaagc aaaaagcaga tcttgtcttc gttgggagtg aattagccct tccagtcccc
3481 ccttttcttt taaaaagtgg ctaagatcta cagctgcctt gtaagtcatt ggtcttaaag
3541 gtaccgagct cgaattccag gcggggaggc ggcccaaagg gagatccgac tcgtctgagg
3601 gcgaaggcga agacgcggaa gaggccgcag agccggcagc aggccgcggg aaggaaggtc
3661 cgctggattg agggccgaag ggacgtagca gaaggacgtc ccgcgcagaa tccaggtggc
3721 aacacaggcg agcagccatg gaaaggacgt cagcttcccc gacaacacca cggaattgtc
3781 agtgcccaac agccgagccc ctgtccagca gcgggcaagg caggcggcga tgagttccgc
3841 cgtggcaata gggaggggga aagcgaaagt cccggaaagg agctgacagg tggtggcaat
3901 gccccaacca gtgggggttg cgtcagcaaa cacagtgcac accacgccac gttgcctgac
3961 aacgggccac aactcctcat aaagagacag caaccaggat ttatacaagg aggagaaaat
4021 gaaagccata cgggaagcaa tagcatgata caaaggcatt aaagcagcgt atccacatag
4081 cgtaaaagga gcaacatagt taagaatacc agtcaatctt tcacaaattt tgtaatccag
4141 aggttgattg tcgagttagg cgccaggctt tcgagtcatg caccaggttc gaggtccctc
4201 aggcacttcc acatcggcgg tgacggtgaa acccaacctt tcatagaaag gaaggttcct
4261 aggtgcactg gtctcaagaa atgcgggaac accagctcgt tcagcggcct ccacgccggg
4321 cagaactacg gcgctgccaa gcccctttcc ctggtgatcg ggggaaacgc ctacggtggc
4381 caggaaccat gcgggctcct taggtcggtg tggcgcgagc aggccttcca tctgctgttg
4441 cgcggccagt ctagatccag agagctcggc cattcttggg ccgatctcgg cgaacaccgc
4501 gccggcttcg acgctctctg gtgtggtcca aacggcgacg gcggctccgt catcggccac
4561 ccacaccttt ccaatgtcga gtcccaccct tgtcaaaaac aattcctgca gttcagtcac
4621 cctctcgatg tgccggtcgg gatccacggt atggcgggtt gcggggtaat cggcaaaggc
4681 agcagccagt gttctcactg ctctggggac atcatcgcga gtcgcgagcc gaacagtagg
4741 cttgtactcg gtcatggtgg cactagaggg gtagttttca cgacacctga aatggaagaa
4801 aaaaactttg aaccactgtc tgaggcttga gaatgaacca agatccaaac tcaaaaaggg
4861 caaattccaa ggagaattac atcaagtgcc aagctggcct aacttcagtc tccacccact
4921 cagtgtgggg aaactccatc gcataaaacc cctcccccca acctaaagac gacgtactcc
4981 aaaagctcga gaactaatcg aggtgcctgg acggcgcccg gtactccgtg gagtcacatg
5041 aagcgacggc tgaggacgga aaggcccttt tcctttgtgt gggtgactca cccgcccgct
5101 ctcccgagcg ccgcgtcctc cattttgagc tccctgcagc agggccggga agcggccatc
5161 tttccgctca cgcaactggt gccgaccggg ccagccttgc cgcccagggc ggggcgatac
5221 acggcggcgc gaggccaggc accagagcag gccggccagc ttgagactac ccccgtccga
5281 ttctcggtgg ccgcgctcgc aggccccgcc tcgccgaaca tgtgcgctgg gacgcacggg
5341 ccccgtcgcc gcccgcggcc ccaaaaaccg aaataccagt gtgcagatct tggcccgcat
5401 ttacaagact atcttgccag aaaaaaagcg tcgcaggata tcatcaaaaa ttttaaatgg
5461 ctagagactt atcgaaagca gcgagacagg cgcgaaggtg ccaccagatt cgcacgcggc
5521 ggccccagcg cccaggccag gcctcaactc aagcacgagg cgaaggggct ccttaagcgc
5581 aaggcctcga actctcccac ccacttccaa cccgaagctc gggatcaaga atcacgtact
5641 gcagccaggg gcgtggaagt aattcaaggc acgcaagggc cataacccgt aaagaggcca
5701 ggcccgcggg aaccacacac ggcacttacc tgtgttctgg cggcaaaccc gttgcgaaaa
5761 agaacgttca cggcgactac tgcacttata tacggttctc ccccaccctc gggaaaaagg
5821 cggagccagt acacgacatc actttcccag tttaccccgc gccaccttct ctaggcaccg
5881 gttcaattgc cgacccctcc ccccaacttc tcggggactg tgggcgatgt gcgctctgcc
5941 cactgacggg caccggagcc tcacgcatgc tcttctccac ctcagtgatg acgagagcgg
6001 gcgggtgagg gggcgggaac gcagcgatct ctgggttcta cgttagtggg agtttaacga
6061 cggtccctgg gattccccaa ggcaggggcg agtccttttg tatgaattac tctcagctcc
6121 ggtcggggcg ggttgggggg ggtggtgacg gggaggcatc gataaaattt tgaattttgt
6181 aatttgtttt tgtaattctt tagtttgtat gtctgttgct attatgtcta ctattctttc
6241 ccctgcactg taccccccaa tccccccttt tcttttaaaa gttggatccg gtaccaaggt
6301 cgggcaggaa gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc
6361 tgttagagag ataattagaa ttaatttgac tgtaaacaca aagatattag tacaaaatac
6421 gtgacgtaga aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat
6481 ggactatcat atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt
6541 gtggaaagga cgaaacaccg agacgattaa tgcgtctcgt tttagagcta gaaatagcaa
6601 gttaaaataa ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg gtgctttttt
6661 tctagtgtta accgataccg tcgagatccg ttcactaatc gaatggatct gtctctgtct
6721 ctctctccac cttcttcttc tattccttcg ggcctgtcgg gtcccctcgg ggttgggagg
6781 tgggtctgaa acgataatgg tgaatatccc tgcctaactc tattcactat agaaagtaca
6841 gcaaaaacta ttcttaaacc taccaagcct cctactatca ttatgaataa ttttatatac
6901 cacagccaat ttgttatgtt aaaccaattc cacaaacttg cccatttatc taattccaat
6961 aattcttgtt cattcttttc ttgctggttt tgcgattctt caattaagga gtgtattaag
7021 cttgtgtaat tgttaatttc tctgtcccac tccatccagg tcgtgtgatt ccaaatctgt
7081 tccagagatt tattactcca actagcattc caaggcacag cagtggtgca aatgagtttt
7141 ccagagcaac cccaaatccc caggagctgt tgatccttta ggtatctttc cacagccagg
7201 attcttgcct ggagctgctt gatgccccag actgtgagtt gcaacagatg ctgttgcgcc
7261 tcaatagccc tcagcaaatt gttctgctgc tgcactatac cagacaataa ttgtctggcc
7321 tgtaccgtca gcgtcattga cgctgcgccc atagtgcttc ctgctgctcc caagaaccca
7381 aggaacaaag ctcctattcc cactgctctt ttttctctct gcaccactct tctctttgcc
7441 ttggtgggtg ctactcctaa tggttcaatt tttactactt tatatttata taattcactt
7501 ctccaattgt ccctcatatc tcctcctcca ggtctgaaga tcagcggccg cttgctgtgc
7561 ggtggtctta cttttgtttt gctcttcctc tatcttgtct aaagcttcct tggtgtcttt
7621 tatctctatc ctttgatgca cacaatagag ggttgctact gtattatata atgatctaag
7681 ttcttctgat cctgtctgaa gggatggttg tagctgtccc agtatttgtc tacagccttc
7741 tgatgtttct aacaggccag gattaactgc gaatcgttct agctccctgc ttgcccatac
7801 tatatgtttt aatttatatt ttttctttcc ccctggcctt aaccgaattt tttcccatcg
7861 cgatctaatt ctcccccgct taatactgac gctctcgcac ccatctctct ccttctagcc
7921 tccgctagtc aaaatttttg gcgtactcac cagtcgccgc ccctcgcctc ttgccgtgcg
7981 cgcttcagca agccgagtcc tgcgtcgaga gagctcctct ggtttccctt tcgctttcaa
8041 gtccctgttc gggcgccact gctagagatt ttccacactg actaaaaggg tctgagggat
8101 ctctagttac cagagtcaca caacagacgg gcacacacta cttgaagcac tcaaggcaag
8161 ctttattgag gcttaagcag tgggttccct agttagccag agagctccca ggctcagatc
8221 tggtctaacc agagagaccc gtttatgtat cgagctaggc acttaaatac aatatctctg
8281 caatgcggca attcagtggt tcgtccaatc catgtcagac ccgtctgttg ccttcctaat
8341 aaggcacgat cgtaccacct tacttccacc aatcggcatg cacggtgctt tttctctcct
8401 tgtaaggcat gttgctaact catcgttacc atgttgcaag actacaagag tattgcataa
8461 gactacatta agctt
//
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