| 质粒名称: | pLV-mTurquoise2-Caspase-1-mNeonGreen |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 122448 |
| 存储实验室: | Feng Zhang |
|
载体骨架基本信息 |
|
| 载体名称: | pcDNA3.1 |
| 载体出品公司: | Clontech |
| 空载体大小: | 4038 bp |
| 完整质粒大小: | 6528bp |
| 载体类型: | 哺乳细胞表达 |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | U6-BhCas12b-sgRNA-scaffold, CMV-mCherry |
|
文献引用方法 |
|
| 材料和方法部分: |
pZ148-BhCas12b-sgRNA-scaffold was a gift from Feng Zhang (Addgene plasmid # 122448 ; http://n2t.net/addgene:122448 ; RRID:Addgene_122448)
|
| 参考文献部分: | Engineering of CRISPR-Cas12b for human genome editing. Strecker J, Jones S, Koopal B, Schmid-Burgk J, Zetsche B, Gao L, Makarova KS, Koonin EV, Zhang F. Nat Commun. 2019 Jan 22;10(1):212. doi: 10.1038/s41467-018-08224-4. 10.1038/s41467-018-08224-4 PubMed 30670702 |
买家导航
pZ148-BhCas12b-sgRNA-scaffold载体质粒基本信息
pZ148-BhCas12b-sgRNA-scaffold质粒图谱载体图谱和pZ148-BhCas12b-sgRNA-scaffold载体序列质粒序列多克隆位点信息
pZ148-BhCas12b-sgRNA-scaffold质粒载体简介
pZ148-BhCas12b-sgRNA-scaffold质粒序列载体序列
LOCUS pZ148-BhCas12b-sgRNA-scaffold 6528 bp DNA circular SYN 14-APR-2025
DEFINITION Expresses sgRNA for BhCas12b in mammalian cells.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6528)
AUTHORS Strecker J, Jones S, Koopal B, Schmid-Burgk J, Zetsche B, Gao L,
Makarova KS, Koonin EV, Zhang F
TITLE Engineering of CRISPR-Cas12b for human genome editing.
JOURNAL Nat Commun. 2019 Jan 22;10(1):212. doi: 10.1038/s41467-018-08224-4.
PUBMED 30670702
REFERENCE 2 (bases 1 to 6528)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Apr 14, 2025 from SnapGene Server 8.0.1
https://www.biofeng.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nat
Commun."; date: "2019-01-22"; volume: "10(1):212. doi"; pages: "
10.1038/s41467-018-08224-4"
COMMENT Sequence Label: pZ148-BhCas12b-sgRNA-scaffold
FEATURES Location/Qualifiers
source 1..6528
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 207..586
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 587..790
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 741..761
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 835..854
/label=T7
/note="T7 promoter, forward primer"
promoter 835..853
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
regulatory 895..904
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 901..1608
/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(1046..1064)
/label=mCherry-R
/note="mCherry, reverse primer"
primer_bind 1314..1333
/label=mCherry-F
/note="mCherry, forward primer"
CDS 1609..1635
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
primer_bind complement(1709..1726)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 1715..1939
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 1985..2413
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2072..2091)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2282..2303
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(2422..2442)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 2427..2756
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 2607..2742
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2669..2688
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 2823..3617
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(2877..2896)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3487..3506
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 3791..3912
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3828..3847)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3882..3901
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(3961..3977)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3961..3977)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(3974..3996)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 3985..4001
/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(4009..4015,4016..4033,4034..4039))
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4054..4075
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4192..4209)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4363..4951)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4443..4462)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(join(5122..5913,5914..5982))
/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 5745..5764
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5983..6087)
/gene="bla"
/label=AmpR promoter
primer_bind complement(6162..6181)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
promoter join(6288..6528,1..8)
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA
(Domitrovich & Kunkel, 2003)"
primer_bind 6288..6308
/label=hU6-F
/note="Human U6 promoter, forward primer"
primer_bind 6459..6478
/label=LKO.1 5'
/note="Human U6 promoter, forward primer"
ORIGIN
1 gaaacaccgg ttctgtcttt tggtcaggac aaccgtctag ctataagtgc tgcagggtgt
61 gagaaactcc tattgctgga cgatgtctct tacgaggcat tagcacggag acgatatatc
121 gtctcttttt ttcaattgca tgaagaatct gcttagggtt aggcgttttg cgctgcttcg
181 cgatgtacgg gccagatata cgcgttgaca ttgattattg actagttatt aatagtaatc
241 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
301 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
361 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
421 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
481 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
541 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg
601 gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc
661 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg
721 taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat
781 aagcagagct ctctggctaa ctagagaacc cactgcttac tggcttatcg aaattaatac
841 gactcactat agggagaccc aagctggcta gcgtttaaac ttaagcttgc cggtgccacc
901 atggtgagca agggcgagga ggataacatg gccatcatca aggagttcat gcgcttcaag
961 gtgcacatgg agggctccgt gaacggccac gagttcgaga tcgagggcga gggcgagggc
1021 cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggtgg ccccctgccc
1081 ttcgcctggg acatcctgtc ccctcagttc atgtacggct ccaaggccta cgtgaagcac
1141 cccgccgaca tccccgacta cttgaagctg tccttccccg agggcttcaa gtgggagcgc
1201 gtgatgaact tcgaggacgg cggcgtggtg accgtgaccc aggactcctc cctgcaggac
1261 ggcgagttca tctacaaggt gaagctgcgc ggcaccaact tcccctccga cggccccgta
1321 atgcagaaga agaccatggg ctgggaggcc tcctccgagc ggatgtaccc cgaggacggc
1381 gccctgaagg gcgagatcaa gcagaggctg aagctgaagg acggcggcca ctacgacgct
1441 gaggtcaaga ccacctacaa ggccaagaag cccgtgcagc tgcccggcgc ctacaacgtc
1501 aacatcaagt tggacatcac ctcccacaac gaggactaca ccatcgtgga acagtacgaa
1561 cgcgccgagg gccgccactc caccggcggc atggacgagc tgtacaagta cccatacgat
1621 gttccagatt acgcttaaga attctgcaga tatccagcac agtggcggcc gctcgagtct
1681 agagggcccg tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
1741 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
1801 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
1861 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
1921 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat
1981 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
2041 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
2101 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
2161 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
2221 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
2281 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
2341 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
2401 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
2461 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
2521 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
2581 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
2641 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct
2701 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
2761 tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc
2821 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
2881 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
2941 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
3001 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
3061 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
3121 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
3181 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
3241 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
3301 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg
3361 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
3421 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
3481 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
3541 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
3601 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
3661 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
3721 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
3781 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
3841 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
3901 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg
3961 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
4021 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
4081 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
4141 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
4201 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
4261 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
4321 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
4381 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
4441 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
4501 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
4561 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
4621 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
4681 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
4741 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
4801 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
4861 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
4921 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
4981 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
5041 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
5101 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
5161 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
5221 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
5281 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
5341 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
5401 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
5461 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
5521 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
5581 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
5641 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
5701 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
5761 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
5821 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
5881 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
5941 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
6001 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
6061 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga
6121 cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc
6181 gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg
6241 agcaaaattt aagctacaac aaggcaaggc ttgaccgact taattaagag ggcctatttc
6301 ccatgattcc ttcatatttg catatacgat acaaggctgt tagagagata attggaatta
6361 atttgactgt aaacacaaag atattagtac aaaatacgtg acgtagaaag taataatttc
6421 ttgggtagtt tgcagtttta aaattatgtt ttaaaatgga ctatcatatg cttaccgtaa
6481 cttgaaagta tttcgatttc ttggctttat atatcttgtg gaaaggac
//
