| 质粒名称: | pPP441 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 158801 |
| 存储实验室: | Jennifer Doudna |
|
载体骨架基本信息 |
|
| 载体名称: | Custom |
| 载体出品公司: | -- |
| 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: | 嘌呤霉素 |
| 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: | CasPhi-2 |
| 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: | -- |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | -- |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pPP441 was a gift from Jennifer Doudna (Addgene plasmid # 158801 ; http://n2t.net/addgene:158801 ; RRID:Addgene_158801) |
| 参考文献部分: | CRISPR-CasPhi from huge phages is a hypercompact genome editor. Pausch P, Al-Shayeb B, Bisom-Rapp E, Tsuchida CA, Li Z, Cress BF, Knott GJ, Jacobsen SE, Banfield JF, Doudna JA. Science. 2020 Jul 17;369(6501):333-337. doi: 10.1126/science.abb1400. 10.1126/science.abb1400 PubMed 32675376 |
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 pPP441 7180 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Plasmid containing Homo sapiens codon optimized Cas-Phi-2 and a
SapI-GG stuffer spacer for editing in human cells..
ACCESSION .
VERSION .
KEYWORDS pPP441
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7180)
AUTHORS Pausch P, Al-Shayeb B, Bisom-Rapp E, Tsuchida CA, Li Z, Cress BF,
Knott GJ, Jacobsen SE, Banfield JF, Doudna JA
TITLE CRISPR-CasPhi from huge phages is a hypercompact genome editor.
JOURNAL Science. 2020 Jul 17;369(6501):333-337. doi:
10.1126/science.abb1400.
PUBMED 32675376
REFERENCE 2 (bases 1 to 7180)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7180
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS 2281..2301
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 2308..2328
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 2347..2394
/codon_start=1
/product="two tandem FLAG(R) epitope tags"
/label=2xFLAG
/translation="DYKDDDDKDYKDDDDK"
CDS 2404..2460
/codon_start=1
/product="2A peptide from porcine teschovirus-1
polyprotein"
/label=P2A
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="ATNFSLLKQAGDVEENPGP"
CDS 2461..3057
/codon_start=1
/gene="pac from Streptomyces alboniger"
/product="puromycin N-acetyltransferase"
/label=PuroR
/note="confers resistance to puromycin"
/translation="TEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIERV
TELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLAA
QQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETSA
PRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
primer_bind 2954..2974
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
primer_bind complement(3091..3108)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 3097..3304
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
repeat_region 3313..3453
/label=AAV2 ITR
/note="inverted terminal repeat of adeno-associated virus
serotype 2"
repeat_region 3313..3442
/label=AAV2 ITR
rep_origin 3528..3983
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3615..3634)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 3825..3846
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(4000..4019)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 4119..4141
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(4179..4197)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 4265..4369
/gene="bla"
/label=AmpR promoter
CDS 4370..5230
/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(4588..4607)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 5401..5989
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5890..5909
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
promoter 6051..6291
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
primer_bind 6051..6071
/label=hU6-F
/note="Human U6 promoter, forward primer"
primer_bind 6222..6241
/label=LKO.1 5'
/note="Human U6 promoter, forward primer"
enhancer 6371..6656
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer;
contains an 18-bp deletion relative to the standard CMV
enhancer"
promoter 6658..6935
/label=chicken beta-actin promoter
intron 6936..7163
/label=hybrid intron
/note="hybrid between chicken beta-actin (CBA) and minute
virus of mice (MMV) introns (Gray et al., 2011)"
ORIGIN
1 atgccaaagc cagccgtgga gtctgagttt tctaaggtac tcaagaagca ctttccgggc
61 gagcgattta ggtctagcta catgaagcgg ggtggtaaaa tcttggcagc ccagggtgaa
121 gaagcggtcg tcgcgtatct gcaaggcaag tccgaggagg aacccccgaa ttttcagccg
181 ccggcgaaat gtcatgttgt tacgaaatca cgagatttcg ccgagtggcc aattatgaag
241 gcctccgaag caatccaaag gtatatctat gcgctctcta cgacggaacg ggcagcttgc
301 aagcctggca aatcttcaga gtcccacgcg gcctggttcg cggcaactgg cgtgtcaaac
361 cacggttata gccatgttca aggcctcaat cttatcttcg accacacgct gggaagatac
421 gatggtgttc tgaaaaaggt gcagctgaga aatgagaaag cccgcgcccg gctggaaagt
481 atcaacgcct ctcgagccga cgaaggactt ccagaaataa aggcagagga ggaagaggtc
541 gctacaaatg aaaccggaca ccttttgcag cctccgggga tcaacccaag tttctacgtt
601 taccagacta tttctccgca ggcttacagg ccgcgagatg agattgtact gccgcccgag
661 tatgccggct acgtccgaga tccgaacgcc cctatccccc ttggcgtggt tcggaatcgg
721 tgcgatattc agaagggatg ccctggatac atccccgaat ggcaaagaga ggcaggtact
781 gcaatttccc ctaagacggg taaagccgtc accgttcccg gcctcagtcc aaaaaaaaat
841 aaacgaatgc gacgatactg gaggtccgag aaagagaagg cccaagatgc actgctcgtt
901 actgtgagaa tcggcactga ctgggtcgta atcgacgttc gaggtttgct gcggaatgcg
961 cggtggcgca ccattgcgcc caaggatata tccttgaatg ccctcttgga tctctttaca
1021 ggcgacccgg tcatagatgt tcggagaaac attgtgactt tcacctacac tctggacgct
1081 tgcggtacat atgctcgcaa atggactctc aaagggaaac agactaaggc aaccctcgat
1141 aagttgaccg caacccagac cgtggccctg gtagcaatag accttggaca aaccaatccc
1201 ataagtgcgg gtatcagtag ggtcacgcaa gaaaacgggg cacttcaatg tgaacctctg
1261 gatcggttca ctctccctga tgatctgctc aaggatatct ccgcgtaccg aatcgcttgg
1321 gatcgcaacg aggaggaact gagggctagg tccgtcgaag cgctcccaga agctcaacaa
1381 gctgaagtga gggctctgga cggcgtttct aaagaaaccg ccaggaccca gctctgcgcg
1441 gacttcggcc ttgatcccaa acggctgcct tgggataaaa tgagcagcaa caccactttc
1501 atcagtgaag cgttgcttag taattctgtg tctagagatc aggttttttt tactcctgcg
1561 cctaaaaagg gagcaaagaa aaaagccccc gttgaagtta tgcggaagga taggacctgg
1621 gcgagggcct ataaaccacg gctcagtgtg gaagcccaaa agctgaaaaa tgaggccttg
1681 tgggctctca agcgcacttc tccagaatac ctcaagctga gtcggagaaa agaggagctt
1741 tgtaggcgaa gtattaacta cgtcattgaa aaaacaagac ggaggacaca atgtcagatc
1801 gtgatacctg tcatagagga cttgaatgtg cgattctttc acggttcagg gaagcgcctg
1861 cctggctggg ataatttttt cactgcgaag aaggagaaca ggtggtttat acagggcctc
1921 cacaaagcat tcagcgactt gcgaactcat cgctccttct acgtattcga agtccgcccg
1981 gagcggactt caataacgtg cccaaaatgc gggcactgcg aggttgggaa ccgggatggg
2041 gaggcttttc agtgccttag ttgcggcaaa acgtgcaatg ccgaccttga cgtggctacc
2101 cataatctga ctcaagtcgc ccttacagga aaaacaatgc cgaaacgcga ggaacctaga
2161 gatgcccagg gcacagctcc agcccgaaaa acaaagaagg cgtcaaagag caaggctccg
2221 ccagccgaac gagaggacca aactccagca caggaaccgt cccagacttc cggaagcgga
2281 cccaagaaaa aacgcaaggt ggaagatcct aagaaaaagc ggaaagtgag cctgggcagc
2341 ggctccgatt acaaagatga cgatgacaaa gactacaagg atgatgatga taagggatcc
2401 ggcgcaacaa acttctctct gctgaaacaa gccggagatg tcgaagagaa tcctggaccg
2461 accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc
2521 accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga tccggaccgc
2581 cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc
2641 ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc
2701 gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc
2761 cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc caaggagccc
2821 gcgtggttcc tggccaccgt cggagtctcg cccgaccacc agggcaaggg tctgggcagc
2881 gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag
2941 acctccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac
3001 gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg tgcctgaacg
3061 cgttaagaat tcctagagct cgctgatcag cctcgactgt gccttctagt tgccagccat
3121 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
3181 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
3241 ggggtggggt ggggcaggac agcaaggggg aggattggga agagaatagc aggcatgctg
3301 gggagcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct
3361 cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct
3421 cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat tttctcctta
3481 cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag
3541 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
3601 cgccttagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
3661 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
3721 cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata
3781 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
3841 aactggaaca acactcaact ctatctcggg ctattctttt gatttataag ggattttgcc
3901 gatttcggtc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
3961 caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc
4021 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
4081 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
4141 gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt
4201 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
4261 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
4321 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
4381 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
4441 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
4501 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
4561 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
4621 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
4681 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
4741 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
4801 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
4861 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
4921 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
4981 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
5041 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtggaagcc gcggtatcat
5101 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
5161 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
5221 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
5281 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
5341 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
5401 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
5461 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
5521 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
5581 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
5641 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
5701 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
5761 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
5821 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
5881 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
5941 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
6001 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt gagggcctat
6061 ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag ataattggaa
6121 ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga aagtaataat
6181 ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat atgcttaccg
6241 taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga cgaaacaccg
6301 gtcggaacgc tcaacgattg cccctcacga ggggacagaa gagctaatgc tcttcatttt
6361 ttttggtacc cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
6421 cccgcccatt gacgtcaata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
6481 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
6541 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttgtgcccag tacatgacct
6601 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtcg
6661 aggtgagccc cacgttctgc ttcactctcc ccatctcccc cccctcccca cccccaattt
6721 tgtatttatt tattttttaa ttattttgtg cagcgatggg ggcggggggg gggggggggc
6781 gcgcgccrgg gggggggggg gggggsgggg ggsggggsgg ggcgaggcgg agaggtgcgg
6841 cggcagccaa tcagagcggc gcgctccgaa agtttccttt tatggcgagg cggcggcggc
6901 ggcggcccta taaaaagcga agcgcgcggc gggcgggagt cgctgcgcgc tgccttcgcc
6961 ccgtgccccg ctccgccgcc gcctcgcgcc gcccgccccg gctctgactg accgcgttac
7021 tcccacaggt gagcgggcgg gacggccctt ctcctccggg ctgtaattag ctgagcaaga
7081 ggtaagggtt taagggatgg ttggttggtg gggtattaat gtttaattac ctggagcacc
7141 tgcctgaaat cacttttttt caggttggac cggtgccacc
//
