| 质粒名称: | pMJ329: pCMV-MmeFz2 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 205266 |
| 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: | 5150bp |
| 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: | MmeFz2 |
| 物种来源: | Mercenaria mercenaria |
| 克隆方法: | Gibson Cloning |
|
文献引用方法 |
|
| 材料和方法部分: | pMJ329: pCMV-MmeFz2 was a gift from Feng Zhang (Addgene plasmid # 205266 ; http://n2t.net/addgene:205266 ; RRID:Addgene_205266) |
| 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 pMJ329: pCMV-MmeFz2 5150 bp ds-DNA circular SYN 09-AUG-2023
DEFINITION Human expression vector for human codon optimized MmeFz2.
ACCESSION .
VERSION .
KEYWORDS pMJ329: pCMV-MmeFz2
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5150)
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 5150)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 9, 2023 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..5150
/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(2096..2115)
/label=Bglob-pA-R
/note="Rabbit beta-globin polyA region, reverse primer"
polyA_signal 2161..2216
/label=-beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal (Gil and
Proudfoot, 1987)"
primer_bind complement(2215..2234)
/label=rbglobpA-R
/note="Rabbit beta-globin polyA, reverse primer. Also
called rb-glob-pA-term-R"
primer_bind complement(2577..2593)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(2577..2593)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(2590..2612)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 2601..2617
/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(2625..2655)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 2670..2691
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 2750..2945
/label=SV40 promoter
/note="SV40 early promoter"
rep_origin 2796..2931
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2858..2877
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
polyA_signal 2951..3085
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2988..3007)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3042..3061
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(3153..3170)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(3324..3912)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3404..3423)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(4083..4943)
/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 4706..4725
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(4944..5048)
/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 tgaagcggaa gcgcgagcag atgacactgt ggaaggccgc
601 cttcgtgaac ggccaagaga cattcaagag ctggatcgac aaggcccgga tgctggaact
661 gaactgcgac gtgtccagcg ccagcagcac acactacagc gacctgaacc tgaaaacaaa
721 gtgcgccaag accgacgaca agttcatgtg caactacagc gtgtgcatca gacccaccag
781 caagcagaag cggaccctga accagatgct gaaggtgtcc aactacgcct acaactggtg
841 caattacctg gtcaaagaga aggacttcaa gcccaagcag ttcgacctgc agcggatcgt
901 ggccaagacc aacagcacag atgtgcccgc cgagtacaga ctgcctggcg acgattggtt
961 cttcgacaac aagatgagca gcatcaagct gaccgcctgc aagaacttct gcaccatgta
1021 caagagcacc cagaccaacc agaaaaagac caaggtggac ctgcggaaca aggacatcgt
1081 gcagctgaga gaaggcagct tcgaggtgca gagcaaatac gtgcggctgc tgaccgagaa
1141 ggacatccct ggcgagagaa tccggcagag cagaatcgcc ctgatgcctg acagcttcag
1201 caagagcaag aaggactgga aagagcggtt cctgcggctg agcaagaacg tgtccaagat
1261 tcctccactg agccacgaca tgaaagtgtg caagcggccc aacggcaagt tcatcctgca
1321 gatcagctgc gaccccatct gcaccagaca gatccaggtg cagaccagcg acagcatctg
1381 cagcattgat cctggcggca gaaccttcgc cacctgttac gaccccagca acatcaagac
1441 cttccagatc ggccctgagg ccgacaagaa agagatcatc cacgagttcc acaacaaaat
1501 cgactacgtg caccggctgc tgtctcacgc ccaagagaag aaacagaccc aggccgtgca
1561 ggacagaatc ggccagctga agaagctgca tctgaagctc aagacctacg tggacgacgt
1621 gcacctgaaa ctgtgcagct acctcgtgaa gaactacaag ctggtggtgc tgggcaagat
1681 cagcgtgtcc tctatcgtgc ggaaggacag acccaaccac ctggccaaga gcgccaacag
1741 agatctgctg tgctggcagc actaccggtt cagacagagg ctgctgcata gagtgcgggg
1801 caccgattgc gaagtgatca tccaggacga gcggtacacc tccaagacct gcggcaattg
1861 cggcgagaag aacaacaaac tcggcggcaa agaaaccttt acctgcgaga gctgcaatta
1921 caagacccac cgggatgtga acggcgccag aaacatcctg tgcaagtatc tgggactgtt
1981 ccccttcgcc gcctgactag cataacccct tggggcgcac tcctcaggtg caggctgcct
2041 atcagaaggt ggtggctggt gtggccaatg ccctggctca caaataccac tgagatcttt
2101 ttccctctgc caaaaattat ggggacatca tgaagcccct tgagcatctg acttctggct
2161 aataaaggaa atttattttc attgcaatag tgtgttggaa ttttttgtgt ctctcactcg
2221 gaaggacata tgggagggca aatcatttaa aacatcagaa tgagtatttg gtttagagtt
2281 tggcaacata tgcccatatg ctggctgcca tgaacaaagg ttggctataa agaggtcatc
2341 agtatatgaa acagccccct gctgtccatt ccttattcca tagaaaagcc ttgacttgag
2401 gttagatttt ttttatattt tgttttgtgt tatttttttc tttaacatcc ctaaaatttt
2461 ccttacatgt tttactagcc agatttttcc tcctctcctg actactccca gtcatagctg
2521 tccctcttct cttatggaga tccctcgacc tgcagcccaa gcttggcgta atcatggtca
2581 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
2641 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
2701 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agcggatccg catctcaatt
2761 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
2821 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
2881 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
2941 gcaaaaagct aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
3001 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
3061 caatgtatct tatcatgtct ggatccgctg cattaatgaa tcggccaacg cgcggggaga
3121 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
3181 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
3241 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
3301 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
3361 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
3421 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
3481 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
3541 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
3601 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
3661 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
3721 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
3781 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
3841 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
3901 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
3961 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
4021 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
4081 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
4141 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
4201 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
4261 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
4321 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
4381 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
4441 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
4501 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
4561 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
4621 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
4681 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
4741 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
4801 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
4861 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
4921 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
4981 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
5041 gttccgcgca catttccccg aaaagtgcca cctgggtcga cattgattat tgactagtaa
5101 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
//
