| 质粒名称: | CMV-enAsCas12f |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 204637 |
| 存储实验室: | Weixin Tang |
|
载体骨架基本信息 |
|
| 载体名称: | de novo assembled |
| 完整质粒大小: | 4776 bp |
| 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: | enAsCas12f / Cas12U3 |
| 物种来源: | Synthetic |
| 基因突变: | D196K, N199K, G276R, N328G, D364R |
| 启动子: | CMV |
| 标签/融合蛋白: |
NLS (N terminal on insert)
NLS (C terminal on insert)
|
| 克隆方法: | 限制性内切酶 |
| 5’测序引物 | CMV-F |
| 3’测序引物 | BGH Reverse |
|
文献引用方法 |
|
| 材料和方法部分: |
CMV-enAsCas12f was a gift from Weixin Tang (Addgene plasmid # 204637 ; http://n2t.net/addgene:204637 ; RRID:Addgene_204637)
|
| 参考文献部分: | An engineered hypercompact CRISPR-Cas12f system with boosted gene-editing activity. Wu T, Liu C, Zou S, Lyu R, Yang B, Yan H, Zhao M, Tang W. Nat Chem Biol. 2023 Jul 3. doi: 10.1038/s41589-023-01380-9. 10.1038/s41589-023-01380-9 PubMed 37400536 |
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 CMV-enAsCas12f 4776 bp ds-DNA circular SYN 09-AUG-2023
DEFINITION Expression of enAsCas12f in mammalian cells.
ACCESSION .
VERSION .
KEYWORDS CMV-enAsCas12f
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4776)
AUTHORS Wu T, Liu C, Zou S, Lyu R, Yang B, Yan H, Zhao M, Tang W
TITLE An engineered hypercompact CRISPR-Cas12f system with boosted
gene-editing activity.
JOURNAL Nat Chem Biol. 2023 Jul 3. doi: 10.1038/s41589-023-01380-9.
PUBMED 37400536
REFERENCE 2 (bases 1 to 4776)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 9, 2023 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..4776
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS complement(59..919)
/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 682..701
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(920..1024)
/gene="bla"
/label=AmpR promoter
primer_bind complement(1103..1122)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 1294..1673
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 1674..1877
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 1828..1848
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 1919..1938
/label=T7
/note="T7 promoter, forward primer"
promoter 1919..1937
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 1987..2007
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 3304..3324
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 3333..3350
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(3373..3390)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 3379..3603
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
primer_bind complement(3674..3690)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3674..3690)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(3687..3709)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 3698..3714
/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(3722..3752)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3767..3788
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(3905..3922)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4076..4664)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4156..4175)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
61 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
121 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
181 gtgctgcaat gataccgcgg ctaccacgct caccggctcc agatttatca gcaataaacc
241 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
301 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
361 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
421 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
481 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
541 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
601 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
661 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
721 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
781 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
841 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
901 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
961 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
1021 cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat cgatctcccg
1081 atcccctagg gtcgactctc agtacaatct gctctgatgc cgcatagtta agccagtatc
1141 tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa
1201 caaggcaagg cttgaccgac aattgcatga agaatctgct tagggttagg cgttttgcgc
1261 tgcttcgcga tgtacgggcc agatatacgc gttgacattg attattgact agttattaat
1321 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac
1381 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
1441 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
1501 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc
1561 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat
1621 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc
1681 ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc
1741 tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa
1801 aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg
1861 tctatataag cagagctggt ttagtgaacc gtcagatccg ctagagatcc gcggccgcta
1921 atacgactca ctatagggag agccgccacc atgaaacgga cagccgacgg aagcgagttc
1981 gagtcaccaa agaagaagcg gaaagtcatg atcaaggtgt acagatacga gatcgtgaag
2041 cccctggacc tggattggaa ggaattcggc accatcctga gacagctgca acaagaaacc
2101 agattcgccc tgaacaaggc cacacagctg gcctgggagt ggatgggctt cagcagcgac
2161 tacaaggaca accatggcga gtaccctaag agcaaggaca tcctgggcta caccaatgtg
2221 cacggctacg cctaccacac aatcaaaaca aaggcctaca gactgaacag cggcaatctg
2281 tcccagacca tcaagcgggc aaccgaccgg tttaaggctt atcagaaaga aatcctgcgg
2341 ggcgacatgt ccatccccag ctacaagcgg gacattcctc tggacctgat caaggagaac
2401 atctctgtga acagaatgaa ccacggcgat tacatcgcca gcctgagcct gctgagcaac
2461 cccgccaagc aggagatgaa cgtgaagcgg aagatcagcg tgatcatcat agttcggggt
2521 gctggaaaga ccatcatgga cagaatcctc agcggcgaat accaggtgtc cgcctctcag
2581 atcatccacg acaagagaaa gaagaaatgg tacctgaata tttcttatga tttcgagcct
2641 cagacaagag tgctggatct caacaagatt atgggcatcg acctgggcgt ggccgtggct
2701 gtgtacatgg cttttcagca cacccctgcc agatacaagc tggaaggcgg agagatcgag
2761 aacttccgta ggcaggtcga gtcccgcaga atcagcatgc tgcggcaggg caagtacgcc
2821 ggcggcgcca gaagaggcca cggcagagat aagcggatca agcctatcga gcagctgaga
2881 gataagatcg ctaacttcag agacaccacc aaccacagat acagccggta catcgtcgac
2941 atggccatca aggagggctg cggcacaatc cagatggaag atctgaccgg catcagagat
3001 attggatcta gattcctgca gaactggacc tactacgacc tgcagcagaa gatcatctac
3061 aaagccgagg aagccggcat caaagtgatc aaaattagac ctcaatacac aagccagcgg
3121 tgcagcgagt gcggcaacat cgacagcgga aatagaatcg gccaggccat cttcaagtgc
3181 agagcgtgtg gatatgaggc caatgccgac tacaacgccg cccgcaacat cgctatccca
3241 aacatcgaca agatcatcgc cgaaagcatc aaaaaaagaa ccgccgacgg cagcgaattc
3301 gagcccaaga agaagaggaa agtctaaccg gtcatcatca ccatcaccat tgagtttaaa
3361 cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc
3421 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
3481 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
3541 acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta
3601 tggcttctga ggcggaaaga accagctggg gctcgatacc gtcgacctct agctagagct
3661 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
3721 acaacatacg agccggaagc ataaagtgta aagcctaggg tgcctaatga gtgagctaac
3781 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
3841 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
3901 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
3961 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
4021 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
4081 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
4141 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
4201 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
4261 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
4321 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
4381 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
4441 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
4501 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
4561 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
4621 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
4681 tttctacggg gtctgacact cagtggaacg aaaactcacg ttaagggatt ttggtcatga
4741 gattatcaaa aaggatcttc acctagatcc ttttaa
//
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