| 质粒名称: | pSLQ1651-sgTelomere(F+E) |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 51024 |
| 存储实验室: | Bo Huang, Stanley Qi |
|
载体骨架基本信息 |
|
| 载体名称: | pSico |
| 载体抗性: | 氨苄霉素 |
| 完整质粒大小: | 8320 bp |
| 载体修饰: | None |
| 载体类型: | 哺乳细胞表达，Lentiviral, CRISPR |
| 筛选标记: | 嘌呤霉素 |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 30 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | Optimized sgRNA |
| 物种来源: | H. sapiens (human), M. musculus (mouse) |
| 插入大小: | 115 bp |
| 基因又名: | sgRNA(F+E) |
| 启动子 | mouse U6 promoter |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | BstXI(未破坏) |
| 3' 克隆位点: | XhoI(未破坏) |
| 5' 测序引物 | atccgacgcgccatctctag |
| 3' 测序引物 | tgcatggcggtaatacggttatc |
|
文献引用方法 |
|
| 材料和方法部分: | pSLQ1651-sgTelomere(F+E) was a gift from Bo Huang & Stanley Qi (爱迪基因 plasmid # 51024) |
| 参考文献部分: | Dynamic Imaging of Genomic Loci in Living Human Cells by an Optimized CRISPR/Cas System. Chen B, Gilbert LA, Cimini BA, Schnitzbauer J, Zhang W, Li GW, Park J, Blackburn EH, Weissman JS, Qi LS, Huang B. Cell. 2013 Dec 19;155(7):1479-91. doi: 10.1016/j.cell.2013.12.001. 10.1016/j.cell.2013.12.001 PubMed 24360272 |
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
质粒图谱.png)
载体简介
载体序列
LOCUS pSLQ1651-sgTelomere(F+E) 8309 bp ds-DNA circular SYN 10-JAN-2018
DEFINITION Lentiviral vector that contains an optimized S. pyogenes sgRNA
targeting human telomeres.
ACCESSION .
VERSION .
KEYWORDS pSLQ1651-sgTelomere(F+E)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8309)
AUTHORS Chen B, Gilbert LA, Cimini BA, Schnitzbauer J, Zhang W, Li GW, Park
J, Blackburn EH, Weissman JS, Qi LS, Huang B
TITLE Dynamic Imaging of Genomic Loci in Living Human Cells by an
Optimized CRISPR/Cas System.
JOURNAL Cell. 2013 Dec 19;155(7):1479-91. doi: 10.1016/j.cell.2013.12.001.
PUBMED 24360272
COMMENT This file is created by Vector NTI
http://www.biofeng.com/
COMMENT ORIGDB|GenBank
COMMENT VNTAUTHORNAME|biofeng.com|
COMMENT VNTNAME|pSLQ1651-sgTelomere(F+E)|
FEATURES Location/Qualifiers
source 1..8309
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(41..60)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 232..611
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 613..811
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
LTR 829..1009
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 1056..1181
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1678..1911
/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 2438..2555
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
protein_bind 2808..2841
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
primer_bind 2845..2865
/label=mU6-F
/note="Mouse U6 promoter, forward primer"
enhancer 3160..3463
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 3464..3667
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
CDS 3701..4297
/codon_start=1
/gene="pac from Streptomyces alboniger"
/product="puromycin N-acetyltransferase"
/label=PuroR
/note="confers resistance to puromycin"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
primer_bind complement(3701..3720)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 4197..4217
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
CDS 4307..4360
/codon_start=1
/product="2A peptide from Thosea asigna virus capsid
protein"
/label=T2A
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="EGRGSLLTCGDVEENPGP"
regulatory 4367..4376
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 4373..5083
/codon_start=1
/product="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
/label=mCherry
/note="mammalian codon-optimized"
/translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
EGRHSTGGMDELYK"
primer_bind complement(4518..4536)
/label=mCherry-R
/note="mCherry, reverse primer"
primer_bind 4786..4805
/label=mCherry-F
/note="mCherry, forward primer"
protein_bind 5103..5136
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
misc_feature 5192..5780
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(5245..5265)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(5663..5674)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
primer_bind complement(5783..5799)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(5784..5800)
/label=pBluescriptKS
/note="For pBluescript vector"
LTR 6309..6489
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
rep_origin complement(6551..7139)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(6631..6650)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(7310..8170)
/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 7933..7952
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(8171..8275)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 ggatcgggag atctcccgat cccctatggt gcactctcag tacaatctgc tctgatgccg
61 catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga
121 gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta
181 gggttaggcg ttttgcgctg cttcgcgatg tacgggccag atatacgcgt tgacattgat
241 tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
301 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
361 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
421 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
481 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
541 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
601 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
661 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
721 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
781 ggcgtgtacg gtgggaggtc tatataagca gcgcgttttg cctgtactgg gtctctctgg
841 ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct
901 caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt
961 aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag tggcgcccga
1021 acagggactt gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg actcggcttg
1081 ctgaagcgcg cacggcaaga ggcgaggggc ggcgactggt gagtacgcca aaaattttga
1141 ctagcggagg ctagaaggag agagatgggt gcgagagcgt cagtattaag cgggggagaa
1201 ttagatcgcg atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa tataaattaa
1261 aacatatagt atgggcaagc agggagctag aacgattcgc agttaatcct ggcctgttag
1321 aaacatcaga aggctgtaga caaatactgg gacagctaca accatccctt cagacaggat
1381 cagaagaact tagatcatta tataatacag tagcaaccct ctattgtgtg catcaaagga
1441 tagagataaa agacaccaag gaagctttag acaagataga ggaagagcaa aacaaaagta
1501 agaccaccgc acagcaagcg gccggccgct gatcttcaga cctggaggag gagatatgag
1561 ggacaattgg agaagtgaat tatataaata taaagtagta aaaattgaac cattaggagt
1621 agcacccacc aaggcaaaga gaagagtggt gcagagagaa aaaagagcag tgggaatagg
1681 agctttgttc cttgggttct tgggagcagc aggaagcact atgggcgcag cgtcaatgac
1741 gctgacggta caggccagac aattattgtc tggtatagtg cagcagcaga acaatttgct
1801 gagggctatt gaggcgcaac agcatctgtt gcaactcaca gtctggggca tcaagcagct
1861 ccaggcaaga atcctggctg tggaaagata cctaaaggat caacagctcc tggggatttg
1921 gggttgctct ggaaaactca tttgcaccac tgctgtgcct tggaatgcta gttggagtaa
1981 taaatctctg gaacagattt ggaatcacac gacctggatg gagtgggaca gagaaattaa
2041 caattacaca agcttaatac actccttaat tgaagaatcg caaaaccagc aagaaaagaa
2101 tgaacaagaa ttattggaat tagataaatg ggcaagtttg tggaattggt ttaacataac
2161 aaattggctg tggtatataa aattattcat aatgatagta ggaggcttgg taggtttaag
2221 aatagttttt gctgtacttt ctatagtgaa tagagttagg cagggatatt caccattatc
2281 gtttcagacc cacctcccaa ccccgagggg acccgacagg cccgaaggaa tagaagaaga
2341 aggtggagag agagacagag acagatccat tcgattagtg aacggatcgg cactgcgtgc
2401 gccaattctg cagacaaatg gcagtattca tccacaattt taaaagaaaa ggggggattg
2461 gggggtacag tgcaggggaa agaatagtag acataatagc aacagacata caaactaaag
2521 aattacaaaa acaaattaca aaaattcaaa attttcgggt ttattacagg gacagcagag
2581 atccagtttg gttagtaccg ggcccgctct agagatccga cgcgccatct ctaggcccgc
2641 gccggccccc tcgcacggac ttgtgggaga agctcggcta ctcccctgcc ccggttaatt
2701 tgcatataat atttcctagt aactatagag gcttaatgtg cgataaaaga cagataatct
2761 gttcttttta atactagcta cattttacat gataggcttg gatttctata acttcgtata
2821 gcatacatta tacgaagtta taaacagcac aaaaggaaac tcaccctaac tgtaaagtaa
2881 ttgtgtgttt tgagactata agtatccctt ggagaaccac cttgttggtt agggttaggg
2941 ttagggttag tttaagagct atgctggaaa cagcatagca agtttaaata aggctagtcc
3001 gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt ttctcgagta ctaggatcca
3061 ttaggcggcc gcgtggataa ccgtattacc gccatgcatt agttattaat agtaatcaat
3121 tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa
3181 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
3241 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
3301 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
3361 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc
3421 tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca
3481 gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat
3541 tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa
3601 caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
3661 cagagctggt ttagtgaacc gtcagatccg ctagcccacc atgaccgagt acaagcccac
3721 ggtgcgcctc gccacccgcg acgacgtccc ccgggccgta cgcaccctcg ccgccgcgtt
3781 cgccgactac cccgccacgc gccacaccgt cgacccggac cgccacatcg agcgggtcac
3841 cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc
3901 ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt
3961 gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca
4021 acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac
4081 cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg
4141 agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa
4201 cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg
4261 accgcgcacc tggtgcatga cccgcaagcc cggtgccgga tcgggagagg gcagaggaag
4321 tctgctaaca tgcggtgacg tcgaggagaa tcctggccca ccggtcgcca ccatggtgag
4381 caagggcgag gaggataaca tggccatcat caaggagttc atgcgcttca aggtgcacat
4441 ggagggctcc gtgaacggcc acgagttcga gatcgagggc gagggcgagg gccgccccta
4501 cgagggcacc cagaccgcca agctgaaggt gaccaagggt ggccccctgc ccttcgcctg
4561 ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc accccgccga
4621 catccccgac tacttgaagc tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa
4681 cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg acggcgagtt
4741 catctacaag gtgaagctgc gcggcaccaa cttcccctcc gacggccccg taatgcagaa
4801 gaagaccatg ggctgggagg cctcctccga gcggatgtac cccgaggacg gcgccctgaa
4861 gggcgagatc aagcagaggc tgaagctgaa ggacggcggc cactacgacg ctgaggtcaa
4921 gaccacctac aaggccaaga agcccgtgca gctgcccggc gcctacaacg tcaacatcaa
4981 gttggacatc acctcccaca acgaggacta caccatcgtg gaacagtacg aacgcgccga
5041 gggccgccac tccaccggcg gcatggacga gctgtacaag taagaattcg tcgagggacc
5101 taataacttc gtatagcata cattatacga agttatacat gtttaagggt tccggttcca
5161 ctaggtacaa ttcgatatca agcttatcga taatcaacct ctggattaca aaatttgtga
5221 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
5281 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
5341 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
5401 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
5461 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
5521 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
5581 ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg
5641 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
5701 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc
5761 cctttgggcc gcctccccgc atcgataccg tcgacctcga tcgagaccta gaaaaacatg
5821 gagcaatcac aagtagcaat acagcagcta ccaatgctga ttgtgcctgg ctagaagcac
5881 aagaggagga ggaggtgggt tttccagtca cacctcaggt acctttaaga ccaatgactt
5941 acaaggcagc tgtagatctt agccactttt taaaagaaaa ggggggactg gaagggctaa
6001 ttcactccca acgaagacaa gatatccttg atctgtggat ctaccacaca caaggctact
6061 tccctgattg gcagaactac acaccagggc cagggatcag atatccactg acctttggat
6121 ggtgctacaa gctagtacca gttgagcaag agaaggtaga agaagccaat gaaggagaga
6181 acacccgctt gttacaccct gtgagcctgc atgggatgga tgacccggag agagaagtat
6241 tagagtggag gtttgacagc cgcctagcat ttcatcacat ggcccgagag ctgcatccgg
6301 actgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag
6361 ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc
6421 gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa
6481 tctctagcag catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
6541 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
6601 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
6661 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
6721 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
6781 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
6841 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
6901 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
6961 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
7021 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
7081 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
7141 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
7201 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
7261 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
7321 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
7381 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
7441 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
7501 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
7561 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
7621 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
7681 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
7741 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
7801 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
7861 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
7921 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
7981 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
8041 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
8101 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
8161 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
8221 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
8281 ttccccgaaa agtgccacct gacgtcgac
//
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