| 质粒名称: | pMSSM24: pCMV-SpuFz1 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 205260 |
| 存储实验室: | Feng Zhang |
|
载体骨架基本信息 |
|
| 载体名称: | pCMV |
| 完整质粒大小: | 5630 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: | SpuFz1 |
| 物种来源: | Spizellomyces punctatus |
| 克隆方法: | Gibson Cloning |
|
文献引用方法 |
|
| 材料和方法部分: | pMSSM24: pCMV-SpuFz1 was a gift from Feng Zhang (Addgene plasmid # 205260 ; http://n2t.net/addgene:205260 ; RRID:Addgene_205260) |
| 参考文献部分: | Fanzor is a eukaryotic programmable RNA-guided endonuclease. Saito M, Xu P, Faure G, Maguire S, Kannan S, Altae-Tran H, Vo S, Desimone A, Macrae RK, Zhang F. Nature. 2023 Jun 28. doi: 10.1038/s41586-023-06356-2. 10.1038/s41586-023-06356-2 PubMed 37380027 |
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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 pMSSM24: pCMV-SpuFz1 5630 bp ds-DNA circular SYN 09-AUG-2023
DEFINITION Human expression vector for human codon optimized SpuFz1.
ACCESSION .
VERSION .
KEYWORDS pMSSM24: pCMV-SpuFz1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5630)
AUTHORS Saito M, Xu P, Faure G, Maguire S, Kannan S, Altae-Tran H, Vo S,
Desimone A, Macrae RK, Zhang F
TITLE Fanzor is a eukaryotic programmable RNA-guided endonuclease.
JOURNAL Nature. 2023 Jun 28. doi: 10.1038/s41586-023-06356-2.
PUBMED 37380027
REFERENCE 2 (bases 1 to 5630)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 9, 2023 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..5630
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 1..304
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 305..508
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 459..479
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind complement(2576..2595)
/label=Bglob-pA-R
/note="Rabbit beta-globin polyA region, reverse primer"
polyA_signal 2641..2696
/label=-beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal (Gil and
Proudfoot, 1987)"
primer_bind complement(2695..2714)
/label=rbglobpA-R
/note="Rabbit beta-globin polyA, reverse primer. Also
called rb-glob-pA-term-R"
primer_bind complement(3057..3073)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3057..3073)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(3070..3092)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 3081..3097
/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(3105..3135)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3150..3171
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 3230..3425
/label=SV40 promoter
/note="SV40 early promoter"
rep_origin 3276..3411
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3338..3357
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
polyA_signal 3431..3565
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3468..3487)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3522..3541
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(3633..3650)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(3804..4392)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3884..3903)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(4563..5423)
/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 5186..5205
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5424..5528)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
541 ccggactcag atcgccacca tgccgcctaa gaagaagcag aagctggaac ggctgaagaa
601 actggacaag cccacactgc acacctgtaa caagaccagc ttcgccaagg ccttcctgcc
661 taacgagaca taccggcagc ggctgctgga ctatatcgcc atcattcacc agctggccga
721 ccacgcctct cacgccctga agttttacat cctgagcacc tccaccagca gcttccctgt
781 ggtgcacgag gataccatcg aggccatcct gtacctgctg aacaagggcg aagcctggca
841 tcccagaaaa gaggccaaga aggcctggcg ggattgcctg ctgccttacg tgcagagata
901 ctgccagatc gtgggcttca ttcaccccaa cctgagaggc gagcagcaga gcatcaacta
961 cctgaccgtg tccatgatga ccaacctgaa agtgaacgtg caagagcact tcatgcagat
1021 gctgctgcgg tacatcaatc tgcgcttcga cgtgaagggc cagaagcaga gactgcctcc
1081 taagagcgac gcccgcaagg cctttttcac ccggctgaga tacctgaagt ccgtgttcct
1141 gttcgacgtg gtgcccgagc tggaattcct ggacgatctg acccctctgg aaagcgaggt
1201 gctggaagag atctggtccc tggacctgcc ttttctgccc aacgatcctc tggcctacgc
1261 catcgtggcc gatcctatga gcttcttccc agcctactgc aagctgagcg gcctgtacga
1321 gcagtacggc ttccagcggt ttagcgccat tcctctgcgg agaagcctga tccagtctca
1381 cgtgcggatc gacaccatca tcctgtatca gcacatcctg tgcatcacca gacgggacgc
1441 cgagacagtg gaaaaggacg acctgtggat gagagtgtgc aacctgtgca ccaaagcctt
1501 ccggtccaga tgcggcatgc acttcgaggg cagcatcaca acagatggcg ccagcgtgtc
1561 cgtgtacctg aaacaccctg aggccgataa gtacggcaag agaggcgcca gaaagtccgc
1621 caatactgtg gccgccgaag tgaaggccct gtacgtggaa aacaacctgc ctgcctgtag
1681 agccgccgag aacgtggtgg tcatcgaccc caacaagcgg gatatcctgt actgccagga
1741 cagcaacggc accaccttca gatacaccgc caaccagaga gccgtggaaa ccggcagcag
1801 aagattcgcc aagcggcggg aagccatgaa ggaagaagcc ggcgttgacc tgatcgagag
1861 cagaatccct agccacaaga ccatgaacct gatggacttc accagatacc tgctcgtgcg
1921 gagagccgac tgggacagac ggaaagagtt ctactctcac cccgctcaca cccggtggaa
1981 gtggcacagc ttcatcaacc ggcagaagtc cgagagcgac ctgatcagca acatgcggaa
2041 caaatacggc gagaacttca ccgtggtcat gggcgattgg agcgacgctg gaagaaccgc
2101 cagattccag accagcagca agaccaaagg ctggcggacc ctgttcaagc ggaaccggat
2161 cgattgcttc ctgctggacg agtacaagac ctccagcgtg tgccctagat gcagcagcag
2221 cgagttcgtg gaaaagaagt tcaagacccg gcctcacagc agaccctggc gaagaagaga
2281 gggcaagatc gagaaggtgc acggcctgct gggctgcacc aatcctaatt gtctgcagca
2341 ggcctggacc agcggcatga gatactggaa ccgggacatg ctgagcacat gcaacatgct
2401 gctgatcgtg cggtccatgc tggatggcca tggcagacct gaggtgttct ctagatctgt
2461 gcctgccgtg gcctgactag cataacccct tggggcgcac tcctcaggtg caggctgcct
2521 atcagaaggt ggtggctggt gtggccaatg ccctggctca caaataccac tgagatcttt
2581 ttccctctgc caaaaattat ggggacatca tgaagcccct tgagcatctg acttctggct
2641 aataaaggaa atttattttc attgcaatag tgtgttggaa ttttttgtgt ctctcactcg
2701 gaaggacata tgggagggca aatcatttaa aacatcagaa tgagtatttg gtttagagtt
2761 tggcaacata tgcccatatg ctggctgcca tgaacaaagg ttggctataa agaggtcatc
2821 agtatatgaa acagccccct gctgtccatt ccttattcca tagaaaagcc ttgacttgag
2881 gttagatttt ttttatattt tgttttgtgt tatttttttc tttaacatcc ctaaaatttt
2941 ccttacatgt tttactagcc agatttttcc tcctctcctg actactccca gtcatagctg
3001 tccctcttct cttatggaga tccctcgacc tgcagcccaa gcttggcgta atcatggtca
3061 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
3121 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
3181 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agcggatccg catctcaatt
3241 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
3301 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
3361 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
3421 gcaaaaagct aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
3481 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
3541 caatgtatct tatcatgtct ggatccgctg cattaatgaa tcggccaacg cgcggggaga
3601 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
3661 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
3721 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
3781 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
3841 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
3901 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
3961 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
4021 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
4081 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
4141 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
4201 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
4261 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
4321 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
4381 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
4441 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
4501 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
4561 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
4621 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
4681 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
4741 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
4801 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
4861 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
4921 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
4981 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
5041 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
5101 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
5161 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
5221 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
5281 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
5341 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
5401 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
5461 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
5521 gttccgcgca catttccccg aaaagtgcca cctgggtcga cattgattat tgactagtaa
5581 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
//
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