| 质粒名称: | PX403 Streptococcus thermophilus LMD-9 CRISPR 3 Cas9 |
|---|---|
| 出品公司： | addgene |
| 目录编号: | 68337 |
| 存储实验室: | Feng Zhang |
|
载体骨架基本信息 |
|
| 载体名称: | 未知 |
| 完整质粒大小: | 8238 bp |
| 载体类型: | 哺乳细胞表达、CRISPR |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | Stbl3 |
| 拷贝数: | 未知 |
|
插入基因信息 |
|
| 插入基因名称: | St3Cas9 / Streptococcus thermophilus LMD-9 CRISPR 3 Cas9 |
| 物种来源: | Synthetic; Streptococcus thermophilus LMD-9 CRISPR 3 |
| 启动子: | CBh |
| 标签/融合蛋白: |
NLS (N terminal on insert)
NLS (C terminal on insert)
HA (N terminal on insert)
|
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | AgeI(未破坏) |
| 3’克隆位点: | EcoRI(未破坏) |
| 5’测序引物 | pCBhProF2 (5'-agggtttaagggatggttgg-3') |
| 3’测序引物 | BGH-rev |
|
文献引用方法 |
|
| 材料和方法部分: | PX403 Streptococcus thermophilus LMD-9 CRISPR 3 Cas9 was a gift from Feng Zhang (Addgene plasmid # 68337 ; http://n2t.net/addgene:68337 ; RRID:Addgene_68337) |
PX403 Streptococcus thermophilus LMD-9 CRISPR 3 Cas9
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 PX403 Streptococcus thermophilus LMD-9 CRISPR 3 Cas9 8238 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Streptococcus thermophilus LMD-9 CRISPR 3 Cas9.
ACCESSION .
VERSION .
KEYWORDS PX403 Streptococcus thermophilus LMD-9 CRISPR 3 Cas9
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8238)
TITLE Feng Zhang lab bacterial Cas9 orthologs
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 8238)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..8238
/organism="synthetic DNA construct"
/mol_type="other DNA"
repeat_region 1..141
/label=AAV2 ITR
/note="inverted terminal repeat of adeno-associated virus
serotype 2"
repeat_region 12..141
/label=AAV2 ITR
enhancer 160..463
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 465..742
/label=chicken beta-actin promoter
intron 743..971
/label=hybrid intron
/note="hybrid between chicken beta-actin (CBA) and minute
virus of mice (MMV) introns (Gray et al., 2011)"
CDS 992..1018
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
CDS 1022..1042
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 5222..5242
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
primer_bind complement(5279..5296)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 5285..5492
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
repeat_region 5501..5641
/label=AAV2 ITR
/note="inverted terminal repeat of adeno-associated virus
serotype 2"
repeat_region 5501..5630
/label=AAV2 ITR
rep_origin 5716..6171
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(5803..5822)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 6013..6034
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(6188..6207)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 6307..6329
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(6367..6385)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 6453..6557
/gene="bla"
/label=AmpR promoter
CDS 6558..7418
/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 complement(6776..6795)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 7589..8177
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 8078..8097
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
121 actccatcac taggggttcc tgcggcctct agaggtaccc gttacataac ttacggtaaa
181 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
241 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
301 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
361 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc
421 tacttggcag tacatctacg tattagtcat cgctattacc atggtcgagg tgagccccac
481 gttctgcttc actctcccca tctccccccc ctccccaccc ccaattttgt atttatttat
541 tttttaatta ttttgtgcag cgatgggggc gggggggggg ggggggcgcg cgccaggcgg
601 ggcggggcgg ggcgaggggc ggggcggggc gaggcggaga ggtgcggcgg cagccaatca
661 gagcggcgcg ctccgaaagt ttccttttat ggcgaggcgg cggcggcggc ggccctataa
721 aaagcgaagc gcgcggcggg cgggagtcgc tgcgacgctg ccttcgcccc gtgccccgct
781 ccgccgccgc ctcgcgccgc ccgccccggc tctgactgac cgcgttactc ccacaggtga
841 gcgggcggga cggcccttct cctccgggct gtaattagct gagcaagagg taagggttta
901 agggatggtt ggttggtggg gtattaatgt ttaattacct ggagcacctg cctgaaatca
961 ctttttttca ggttggaccg gtgccaccat gtacccatac gatgttccag attacgcttc
1021 gccgaagaaa aagcgcaagg tcgaagcgtc caccaagccc tacagcatcg gcctggacat
1081 cggcaccaat agcgtgggct gggccgtgac caccgacaac tacaaggtgc ccagcaagaa
1141 aatgaaggtg ctgggcaaca cctccaagaa gtacatcaag aaaaacctgc tgggcgtgct
1201 gctgttcgac agcggcatta cagccgaggg cagacggctg aagagaaccg ccagacggcg
1261 gtacacccgg cggagaaaca gaatcctgta tctgcaagag atcttcagca ccgagatggc
1321 taccctggac gacgccttct tccagcggct ggacgacagc ttcctggtgc ccgacgacaa
1381 gcgggacagc aagtacccca tcttcggcaa cctggtggaa gagaaggcct accacgacga
1441 gttccccacc atctaccacc tgagaaagta cctggccgac agcaccaaga aggccgacct
1501 gagactggtg tatctggccc tggcccacat gatcaagtac cggggccact tcctgatcga
1561 gggcgagttc aacagcaaga acaacgacat ccagaagaac ttccaggact tcctggacac
1621 ctacaacgcc atcttcgaga gcgacctgtc cctggaaaac agcaagcagc tggaagagat
1681 cgtgaaggac aagatcagca agctggaaaa gaaggaccgc atcctgaagc tgttccccgg
1741 cgagaagaac agcggaatct tcagcgagtt tctgaagctg atcgtgggca accaggccga
1801 cttcagaaag tgcttcaacc tggacgagaa agccagcctg cacttcagca aagagagcta
1861 cgacgaggac ctggaaaccc tgctgggata tatcggcgac gactacagcg acgtgttcct
1921 gaaggccaag aagctgtacg acgctatcct gctgagcggc ttcctgaccg tgaccgacaa
1981 cgagacagag gccccactga gcagcgccat gattaagcgg tacaacgagc acaaagagga
2041 tctggctctg ctgaaagagt acatccggaa catcagcctg aaaacctaca atgaggtgtt
2101 caaggacgac accaagaacg gctacgccgg ctacatcgac ggcaagacca accaggaaga
2161 ggaagatttc tatgtgtacc tgaagaagct gctggccgag ttcgaggggg ccgactactt
2221 tctggaaaaa atcgaccgcg aggatttcct gcggaagcag cggaccttcg acaacggcag
2281 catcccctac cagatccatc tgcaggaaat gcgggccatc ctggacaagc aggccaagtt
2341 ctacccattc ctggccaaga acaaagagcg gatcgagaag atcctgacct tccgcatccc
2401 ttactacgtg ggccccctgg ccagaggcaa cagcgatttt gcctggtcca tccggaagcg
2461 caatgagaag atcaccccct ggaacttcga ggacgtgatc gacaaagagt ccagcgccga
2521 ggccttcatc aaccggatga ccagcttcga cctgtacctg cccgaggaaa aggtgctgcc
2581 caagcacagc ctgctgtacg agacattcaa tgtgtataac gagctgacca aagtgcggtt
2641 tatcgccgag tctatgcggg actaccagtt cctggactcc aagcagaaaa aggacatcgt
2701 gcggctgtac ttcaaggaca agcggaaagt gaccgataag gacatcatcg agtacctgca
2761 cgccatctac ggctacgatg gcatcgagct gaagggcatc gagaagcagt tcaactccag
2821 cctgagcaca taccacgacc tgctgaacat tatcaacgac aaagaatttc tggacgactc
2881 cagcaacgag gccatcatcg aagagatcat ccacaccctg accatctttg aggaccgcga
2941 gatgatcaag cagcggctga gcaagttcga gaacatcttc gacaagagcg tgctgaaaaa
3001 gctgagcaga cggcactaca ccggctgggg caagctgagc gccaagctga tcaacggcat
3061 ccgggacgag aagtccggca acacaatcct ggactacctg atcgacgacg gcatcagcaa
3121 ccggaacttc atgcagctga tccacgacga cgccctgagc ttcaagaaga agatccagaa
3181 ggcccagatc atcggggacg aggacaaggg caacatcaaa gaagtcgtga agtccctgcc
3241 cggcagcccc gccatcaaga agggaatcct gcagagcatc aagatcgtgg acgagctcgt
3301 gaaagtgatg ggcggcagaa agcccgagag catcgtggtg gaaatggcta gagagaacca
3361 gtacaccaat cagggcaaga gcaacagcca gcagagactg aagagactgg aaaagtccct
3421 gaaagagctg ggcagcaaga ttctgaaaga gaatatccct gccaagctgt ccaagatcga
3481 caacaacgcc ctgcagaacg accggctgta cctgtactac ctgcagaatg gcaaggacat
3541 gtatacaggc gacgacctgg atatcgaccg cctgagcaac tacgacatcg accatattat
3601 cccccaggcc ttcctgaaag acaacagcat tgacaacaaa gtgctggtgt cctccgccag
3661 caaccgcggc aagtccgatg atgtgcccag cctggaagtc gtgaaaaaga gaaagacctt
3721 ctggtatcag ctgctgaaaa gcaagctgat tagccagagg aagttcgaca acctgaccaa
3781 ggccgagaga ggcggcctga gccctgaaga taaggccggc ttcatccaga gacagctggt
3841 ggaaacccgg cagatcacca agcacgtggc cagactgctg gatgagaagt ttaacaacaa
3901 gaaggacgag aacaaccggg ccgtgcggac cgtgaagatc atcaccctga agtccaccct
3961 ggtgtcccag ttccggaagg acttcgagct gtataaagtg cgcgagatca atgactttca
4021 ccacgcccac gacgcctacc tgaatgccgt ggtggcttcc gccctgctga agaagtaccc
4081 taagctggaa cccgagttcg tgtacggcga ctaccccaag tacaactcct tcagagagcg
4141 gaagtccgcc accgagaagg tgtacttcta ctccaacatc atgaatatct ttaagaagtc
4201 catctccctg gccgatggca gagtgatcga gcggcccctg atcgaagtga acgaagagac
4261 aggcgagagc gtgtggaaca aagaaagcga cctggccacc gtgcggcggg tgctgagtta
4321 tcctcaagtg aatgtcgtga agaaggtgga agaacagaac cacggcctgg atcggggcaa
4381 gcccaagggc ctgttcaacg ccaacctgtc cagcaagcct aagcccaact ccaacgagaa
4441 tctcgtgggg gccaaagagt acctggaccc taagaagtac ggcggatacg ccggcatctc
4501 caatagcttc accgtgctcg tgaagggcac aatcgagaag ggcgctaaga aaaagatcac
4561 aaacgtgctg gaatttcagg ggatctctat cctggaccgg atcaactacc ggaaggataa
4621 gctgaacttt ctgctggaaa aaggctacaa ggacattgag ctgattatcg agctgcctaa
4681 gtactccctg ttcgaactga gcgacggctc cagacggatg ctggcctcca tcctgtccac
4741 caacaacaag cggggcgaga tccacaaggg aaaccagatc ttcctgagcc agaaatttgt
4801 gaaactgctg taccacgcca agcggatctc caacaccatc aatgagaacc accggaaata
4861 cgtggaaaac cacaagaaag agtttgagga actgttctac tacatcctgg agttcaacga
4921 gaactatgtg ggagccaaga agaacggcaa actgctgaac tccgccttcc agagctggca
4981 gaaccacagc atcgacgagc tgtgcagctc cttcatcggc cctaccggca gcgagcggaa
5041 gggactgttt gagctgacct ccagaggctc tgccgccgac tttgagttcc tgggagtgaa
5101 gatcccccgg tacagagact acaccccctc tagtctgctg aaggacgcca ccctgatcca
5161 ccagagcgtg accggcctgt acgaaacccg gatcgacctg gctaagctgg gcgagggaag
5221 ccccaagaag aagagaaagg tggaggccag ctaagaattc ctagagctcg ctgatcagcc
5281 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
5341 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
5401 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
5461 gattgggaag agaatagcag gcatgctggg gagcggccgc aggaacccct agtgatggag
5521 ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc
5581 cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag
5641 gggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatacgt
5701 caaagcaacc atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
5761 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
5821 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
5881 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg
5941 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
6001 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcgggct
6061 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
6121 tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt ttatggtgca
6181 ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac
6241 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga
6301 ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac
6361 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
6421 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
6481 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
6541 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
6601 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
6661 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
6721 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
6781 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
6841 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
6901 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
6961 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
7021 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
7081 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
7141 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
7201 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
7261 ccggtgagcg tggaagccgc ggtatcattg cagcactggg gccagatggt aagccctccc
7321 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
7381 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
7441 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
7501 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
7561 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
7621 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
7681 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
7741 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
7801 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
7861 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
7921 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
7981 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
8041 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
8101 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
8161 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
8221 ggccttttgc tcacatgt
//
