| 质粒名称: | pAC1453_pX-sgTelomere-25xPBSa |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 73695 |
| 存储实验室: | Albert Cheng |
|
载体骨架基本信息 |
|
| 载体名称: | pX330; pUC ori vector |
| 载体类型: | 哺乳细胞表达,CRISPR |
| 载体抗性: | 氨苄霉素100 μg/mL |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
|
插入基因信息 |
|
| 插入基因名称: | sgTelomere-25xPBSa |
| 启动子: | U6 |
| 5’测序引物 | gagggcctatttcccatgattcc |
| 3’测序引物 | gccatttgtctgcagaattggc |
|
文献引用方法 |
|
| 材料和方法部分: | pAC1453_pX-sgTelomere-25xPBSa was a gift from Albert Cheng (Addgene plasmid # 73695 ; http://n2t.net/addgene:73695 ; RRID:Addgene_73695) |
| 参考文献部分: | Casilio: a versatile CRISPR-Cas9-Pumilio hybrid for gene regulation and genomic labeling. Cheng AW, Jillette N, Lee P, Plaskon D, Fujiwara Y, Wang W, Taghbalout A, Wang H. Cell Res. 2016 Feb;26(2):254-7. doi: 10.1038/cr.2016.3. Epub 2016 Jan 15. 10.1038/cr.2016.3 PubMed 26768771 |
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载体基本信息
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载体简介
载体序列
LOCUS pAC1453_pX-sgTelomere-25xPBSa 3498 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Vector expressing sgTelomere-25xPBSa.
ACCESSION .
VERSION .
KEYWORDS pAC1453_pX-sgTelomere-25xPBSa
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3498)
AUTHORS Cheng AW, Jillette N, Lee P, Plaskon D, Fujiwara Y, Wang W,
Taghbalout A, Wang H
TITLE Casilio: a versatile CRISPR-Cas9-Pumilio hybrid for gene regulation
and genomic labeling.
JOURNAL Cell Res. 2016 Feb;26(2):254-7. doi: 10.1038/cr.2016.3. Epub 2016
Jan 15.
PUBMED 26768771
REFERENCE 2 (bases 1 to 3498)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..3498
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 9..249
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
primer_bind 9..29
/label=hU6-F
/note="Human U6 promoter, forward primer"
primer_bind 180..199
/label=LKO.1 5'
/note="Human U6 promoter, forward primer"
repeat_region 761..901
/label=AAV2 ITR
/note="inverted terminal repeat of adeno-associated virus
serotype 2"
repeat_region 761..890
/label=AAV2 ITR
rep_origin 976..1431
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(1063..1082)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 1273..1294
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(1448..1467)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 1567..1589
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(1627..1645)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 1713..1817
/gene="bla"
/label=AmpR promoter
CDS 1818..2678
/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 complement(2036..2055)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 2849..3437
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 3338..3357
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 ggcgcgccga gggcctattt cccatgattc cttcatattt gcatatacga tacaaggctg
61 ttagagagat aattggaatt aatttgactg taaacacaaa gatattagta caaaatacgt
121 gacgtagaaa gtaataattt cttgggtagt ttgcagtttt aaaattatgt tttaaaatgg
181 actatcatat gcttaccgta acttgaaagt atttcgattt cttggcttta tatatcttgt
241 ggaaaggacg aaacaccgtt agggttaggg ttagggttag tttaagagct atgctggaaa
301 cagcatagca agtttaaata aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc
361 ggtgccaatt gggtctccag attgtatgta gcctgtatgt agcctgtatg tagcctgtat
421 gtagcctgta tgtaagattg tatgtagcct gtatgtagcc tgtatgtagc ctgtatgtag
481 cctgtatgta agattgtatg tagcctgtat gtagcctgta tgtagcctgt atgtagcctg
541 tatgtaagat tgtatgtagc ctgtatgtag cctgtatgta gcctgtatgt agcctgtatg
601 taagattgta tgtagcctgt atgtagcctg tatgtagcct gtatgtagcc tgtatgtaag
661 atcttttttt gttttagagc tagaaatagc aagttaaaat aaggctagtc cgtagcgcgt
721 gcgccaattc tgcagacaaa tggcggcgcg ccgcggccgc aggaacccct agtgatggag
781 ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc
841 cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag
901 gggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatacgt
961 caaagcaacc atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
1021 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
1081 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
1141 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg
1201 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
1261 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcgggct
1321 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
1381 tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt ttatggtgca
1441 ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac
1501 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga
1561 ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac
1621 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
1681 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
1741 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
1801 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
1861 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
1921 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
1981 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
2041 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
2101 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
2161 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
2221 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
2281 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
2341 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
2401 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
2461 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
2521 ccggtgagcg tggaagccgc ggtatcattg cagcactggg gccagatggt aagccctccc
2581 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
2641 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
2701 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
2761 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
2821 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
2881 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
2941 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
3001 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
3061 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
3121 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
3181 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
3241 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
3301 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
3361 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
3421 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
3481 ggccttttgc tcacatgt
//
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