| 质粒名称: | SlugABEmax |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 163798 |
| 存储实验室: | Yongming Wang |
|
载体骨架基本信息 |
|
| 载体名称: | Expression vector pKK44 |
| 载体出品公司: | -- |
| 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,CRISPR |
| 筛选标记: | -- |
| 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: | -- |
| 物种来源: | -- |
| 插入大小: | -- |
| 基因突变: | SlugCas9 (D10A) |
| 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: | SlugABEmax was a gift from Yongming Wang (Addgene plasmid # 163798 ; http://n2t.net/addgene:163798 ; RRID:Addgene_163798) |
| 参考文献部分: | Discovery and engineering of small SlugCas9 with broad targeting range and high specificity and activity. Hu Z, Zhang C, Wang S, Gao S, Wei J, Li M, Hou L, Mao H, Wei Y, Qi T, Liu H, Liu D, Lan F, Lu D, Wang H, Li J, Wang Y. Nucleic Acids Res. 2021 Mar 15. pii: 6171296. doi: 10.1093/nar/gkab148. 10.1093/nar/gkab148 PubMed 33721016 |
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 SlugABEmax 8685 bp ds-DNA circular SYN 13-MAY-2021
DEFINITION Expresses ABEmax and nSlugCas9.
ACCESSION .
VERSION .
KEYWORDS SlugABEmax
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8685)
AUTHORS Hu Z, Zhang C, Wang S, Gao S, Wei J, Li M, Hou L, Mao H, Wei Y, Qi
T, Liu H, Liu D, Lan F, Lu D, Wang H, Li J, Wang Y
TITLE Discovery and engineering of small SlugCas9 with broad targeting
range and high specificity and activity.
JOURNAL Nucleic Acids Res. 2021 Mar 15. pii: 6171296. doi:
10.1093/nar/gkab148.
PUBMED 33721016
REFERENCE 2 (bases 1 to 8685)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..8685
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 200..403
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 354..374
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 445..464
/label=T7
/note="T7 promoter, forward primer"
promoter 445..463
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 513..533
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 4923..4943
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 4953..5009
/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 5010..5726
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(5055..5076)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(5316..5335)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 5663..5684
/label=EGFP-C
/note="EGFP, forward primer"
CDS 5739..5759
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 5768..5785
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(5808..5825)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 5814..6038
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
primer_bind complement(6109..6125)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(6109..6125)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(6122..6144)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 6133..6149
/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(6157..6187)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 6202..6223
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(6340..6357)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(6511..7099)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(6591..6610)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(7270..8130)
/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 7893..7912
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(8131..8235)
/gene="bla"
/label=AmpR promoter
primer_bind complement(8314..8333)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 8505..199
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
ORIGIN
1 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
61 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
121 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
181 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
241 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
301 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
361 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca
421 gatccgctag agatccgcgg ccgctaatac gactcactat agggagagcc gccaccatga
481 aacggacagc cgacggaagc gagttcgagt caccaaagaa gaagcggaaa gtctctgaag
541 tcgagtttag ccacgagtat tggatgaggc acgcactgac cctggcaaag cgagcatggg
601 atgaaagaga agtccccgtg ggcgccgtgc tggtgcacaa caatagagtg atcggagagg
661 gatggaacag gccaatcggc cgccacgacc ctaccgcaca cgcagagatc atggcactga
721 ggcagggagg cctggtcatg cagaattacc gcctgatcga tgccaccctg tatgtgacac
781 tggagccatg cgtgatgtgc gcaggagcaa tgatccacag caggatcgga agagtggtgt
841 tcggagcacg ggacgccaag accggcgcag caggctccct gatggatgtg ctgcaccacc
901 ccggcatgaa ccaccgggtg gagatcacag agggaatcct ggcagacgag tgcgccgccc
961 tgctgagcga tttctttaga atgcggagac aggagatcaa ggcccagaag aaggcacaga
1021 gctccaccga ctctggagga tctagcggag gatcctctgg aagcgagaca ccaggcacaa
1081 gcgagtccgc cacaccagag agctccggcg gctcctccgg aggatcctct gaggtggagt
1141 tttcccacga gtactggatg agacatgccc tgaccctggc caagagggca cgcgatgaga
1201 gggaggtgcc tgtgggagcc gtgctggtgc tgaacaatag agtgatcggc gagggctgga
1261 acagagccat cggcctgcac gacccaacag cccatgccga aattatggcc ctgagacagg
1321 gcggcctggt catgcagaac tacagactga ttgacgccac cctgtacgtg acattcgagc
1381 cttgcgtgat gtgcgccggc gccatgatcc actctaggat cggccgcgtg gtgtttggcg
1441 tgaggaacgc aaaaaccggc gccgcaggct ccctgatgga cgtgctgcac taccccggca
1501 tgaatcaccg cgtcgaaatt accgagggaa tcctggcaga tgaatgtgcc gccctgctgt
1561 gctatttctt tcggatgcct agacaggtgt tcaatgctca gaagaaggcc cagagctcca
1621 ccgactccgg aggatctagc ggaggctcct ctggctctga gacacctggc acaagcgaga
1681 gcgcaacacc tgaaagcagc gggggcagca gcggggggtc aaaccaaaaa ttcatactgg
1741 gactggccat cggaatcacc agcgtgggct acggcctgat cgactacgag acaaagaata
1801 tcatcgatgc cggcgttaga ctgttccccg aggccaacgt ggaaaacaac gagggaagaa
1861 ggtccaaacg tggaagcaga agactgaagc gacgccgcat tcacagactt gaacgggtga
1921 agaagctgct cgaggattat aatctgctgg atcagtccca gattcctcag tctacaaacc
1981 cctacgccat ccgcgtgaag ggcctgtctg aagccctgag caaggacgaa ctcgtgattg
2041 ccctgctcca tatcgccaag agaagaggca tccacaagat cgacgtgatc gacagcaacg
2101 acgacgtggg gaacgagctc agcaccaagg aacagctgaa taagaacagc aagctgctga
2161 aagacaaatt tgtgtgccag atccagctgg aaagaatgaa tgagggccag gtgcggggag
2221 agaaaaaccg gttcaagacc gctgatatca tcaaggaaat catccagctg ctgaatgtgc
2281 agaagaactt ccaccagctg gacgagaact tcatcaacaa gtacatcgaa ctggttgaga
2341 tgagacggga atacttcgag ggccccggca agggcagtcc atatggctgg gaaggcgacc
2401 ctaaggcttg gtacgagaca ctgatgggcc actgcaccta cttcccagat gagctgagaa
2461 gcgtgaaata cgcctacagt gccgacctgt tcaacgctct gaacgacctg aacaacctgg
2521 tcatccaaag agatggactg tctaagctcg agtatcatga gaagtatcac atcatcgaga
2581 acgtgttcaa gcagaagaag aaacctacac tgaagcagat cgccaatgag atcaatgtca
2641 accctgaaga tatcaagggc tacagaatca caaagtctgg caagccccag tttaccgagt
2701 ttaagctcta ccacgacctg aaaagcgtgc tgtttgacca gagcatcctg gagaacgaag
2761 acgtgctgga ccagatcgct gagatcctga ccatctacca ggacaaggat agcatcaaat
2821 ctaagctgac ggaactggac atcctgctga acgaggaaga taaggaaaac atcgcccagc
2881 tgactggcta caccgggacc caccggctca gcctgaaatg catccggctg gtcctggaag
2941 agcagtggta ttctagccgg aatcagatgg aaatcttcac acacctgaac attaagccta
3001 agaagatcaa cctgacagcc gccaacaaga tcccgaaggc tatgatcgac gagttcatcc
3061 tgagccctgt ggtgaagagg accttcggcc aggccattaa ccttattaac aagatcatag
3121 aaaagtacgg cgtgcctgaa gatatcatca tcgagctggc cagagaaaat aatagcaagg
3181 acaagcagaa gttcatcaat gagatgcaga aaaagaacga gaacaccaga aagagaatta
3241 acgaaatcat cggcaagtat ggcaaccaga acgccaagag actggtcgag aagattagac
3301 tgcacgacga gcaggagggc aagtgcctgt actcactgga aagcatccct ctggaggacc
3361 tgctgaacaa ccccaaccac tacgaggtgg accacatcat tccaagatct gtgtccttcg
3421 acaactctta ccacaacaaa gtgctcgtga agcagagcga gaactccaaa aaatccaacc
3481 tgacccctta ccagtacttt aacagcggca agtccaagct ctcttacaac cagtttaaac
3541 aacacatcct gaacctgagc aagtcccagg atagaatcag caaaaaaaag aaagagtatc
3601 tgctggaaga acgggacatc aacaagttcg aggtgcaaaa agagttcatc aatagaaacc
3661 tggtggatac ccggtacgcc acaagagagc tgacaaacta cctgaaggcc tacttcagcg
3721 ccaacaatat gaacgtgaag gtgaaaacga tcaacggcag cttcaccgat tacctgcgga
3781 aagtgtggaa gtttaagaag gaacggaacc acggctacaa gcaccacgcc gaggacgccc
3841 tgattatcgc taatgccgat ttcctgttca aagagaacaa gaagctgaaa gccgtgaact
3901 ctgtgctgga aaaacctgag atcgagagca agcagctgga tatccaggtg gatagcgagg
3961 ataactacag cgaaatgttc atcatcccta agcaggtcca ggacatcaag gacttcagaa
4021 acttcaagta cagccacaga gtggacaaga agcctaacag acagctgatc aacgatacac
4081 tgtacagcac ccggaagaag gacaactcca cctacatcgt gcagaccatc aaagatatct
4141 atgccaaaga taataccacc ctgaagaagc agtttgacaa gtcacccgag aagttcctca
4201 tgtaccaaca cgatccgcgg accttcgaga agttggaagt gatcatgaag cagtacgcta
4261 atgagaagaa tcctctggcc aagtaccacg aggaaacagg cgagtacctg accaaataca
4321 gcaaaaaaaa caacggccct atcgtgaaaa gcctgaagta cattggaaac aagctgggca
4381 gccacctaga tgtgacccac cagttcaaga gcagcaccaa gaagttggtg aagctgagca
4441 tcaagcctta tagattcgac gtctacctga ccgacaaggg atataagttc atcaccatca
4501 gctacctgga cgtgctgaag aaagacaatt actactacat acccgaacag aagtacgaca
4561 agctcaaact gggcaaggcc atcgacaaaa acgccaagtt tatcgctagc ttctacaaga
4621 atgatctgat caagctggac ggcgagatct acaagatcat cggcgtgaat agcgacacca
4681 gaaacatgat cgaactggat ctgcctgaca tcagatacaa agaatactgc gagctgaaca
4741 atatcaaggg cgaacctaga atcaaaaaga ccatcggcaa aaaggtgaat agcatcgaaa
4801 aactgacaac cgacgtgctg ggcaacgtgt tcaccaacac ccagtacaca aaacctcagc
4861 tgctgttcaa gcgaggaaat tctggcggct caaaaagaac cgccgacggc agcgaattcg
4921 agcccaagaa gaagaggaaa gtcggaagcg gagctactaa cttcagcctg ctgaagcagg
4981 ctggagacgt ggaggagaac cctggaccta tggtgagcaa gggcgaggag ctgttcaccg
5041 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt
5101 ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca
5161 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctat ggagtgcagt
5221 gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
5281 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg
5341 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact
5401 tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg
5461 tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca
5521 acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg
5581 acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag
5641 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca
5701 ctctcggcat ggacgagctg tacaagtctg gtggttctcc caagaagaag aggaaagtct
5761 aaccggtcat catcaccatc accattgagt ttaaacccgc tgatcagcct cgactgtgcc
5821 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
5881 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
5941 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
6001 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag
6061 ctggggctcg ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
6121 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
6181 gtgtaaagcc tagggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
6241 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
6301 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
6361 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
6421 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
6481 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
6541 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
6601 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
6661 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
6721 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
6781 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
6841 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
6901 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
6961 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
7021 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
7081 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acactcagtg
7141 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
7201 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
7261 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
7321 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
7381 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
7441 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
7501 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
7561 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
7621 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
7681 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
7741 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
7801 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
7861 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
7921 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
7981 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
8041 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
8101 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
8161 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
8221 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga
8281 tcgatctccc gatcccctag ggtcgactct cagtacaatc tgctctgatg ccgcatagtt
8341 aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat
8401 ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagggttag
8461 gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt gattattgac
8521 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
8581 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
8641 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggac
//
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