| 质粒名称: | pACRISPR |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 113348 |
| 存储实验室: | Quanjiang Ji |
|
载体骨架基本信息 |
|
| 载体名称: | -- |
| 载体出品公司: | -- |
| 载体抗性: | 羧苄青霉素 |
| 空载体大小: | 6936 bp |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | CRISPR,合成生物学 |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | -- |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | -- |
| 物种来源: | -- |
| 插入大小: | -- |
| 基因突变: | -- |
| 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: | pACRISPR was a gift from Quanjiang Ji (Addgene plasmid # 113348 ; http://n2t.net/addgene:113348 ; RRID:Addgene_113348) |
| 参考文献部分: | CRISPR/Cas9-based Genome Editing in Pseudomonas aeruginosa and Cytidine Deaminase-Mediated Base Editing in Pseudomonas Species. Chen W, Zhang Y, Zhang Y, Pi Y, Gu T, Song L, Wang Y, Ji Q. iScience. 2018 Aug 31;6:222-231. doi: 10.1016/j.isci.2018.07.024. Epub 2018 Aug 1. 10.1016/j.isci.2018.07.024 PubMed 30240613 |
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 pACRISPR 6935 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION A sgRNA expression plasmid for genome editing in Pseudomonas
aeruginosa.
ACCESSION .
VERSION .
KEYWORDS pACRISPR
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6935)
AUTHORS Chen W, Zhang Y, Zhang Y, Pi Y, Gu T, Song L, Wang Y, Ji Q
TITLE CRISPR/Cas9-based Genome Editing in Pseudomonas aeruginosa and
Cytidine Deaminase-Mediated Base Editing in Pseudomonas Species.
JOURNAL iScience. 2018 Aug 31;6:222-231. doi: 10.1016/j.isci.2018.07.024.
Epub 2018 Aug 1.
PUBMED 30240613
REFERENCE 2 (bases 1 to 6935)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6935
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(16..37)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 228..247
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 460..482
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 474..491
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 475..491
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 498..517
/label=T7
/note="T7 promoter, forward primer"
promoter 498..516
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind 546..565
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
CDS complement(604..2025)
/codon_start=1
/gene="Bacillus subtilis sacB"
/product="secreted levansucrase that renders bacterial
growth sensitive to sucrose"
/label=SacB
/note="negative selection marker"
/translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
VKDSILEQGQLTVNK"
regulatory 2462..2471
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
promoter 2487..2516
/label=trc promoter
/note="strong E. coli promoter; hybrid between the trp and
lac UV5 promoters"
misc_RNA 2539..2614
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
promoter complement(2694..2712)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(2695..2712)
/label=SP6
/note="SP6 promoter, forward primer"
primer_bind complement(2730..2746)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(2730..2746)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(2743..2765)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 2754..2770
/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(2778..2808)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 2823..2844
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(2915..2933)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 3001..3105
/gene="bla"
/label=AmpR promoter
CDS 3106..3966
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(3324..3343)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 4304..4655
/label=pRO1600 oriV
/note="broad-host-range origin of replication from
Pseudomonas aeruginosa plasmid pRO1600; requires the
pRO1600 Rep protein for replication (West et al., 1994)"
rep_origin 5959..6547
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6448..6467
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 6701..6718
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(6814..334)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
ORIGIN
1 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
61 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa
121 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
181 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
241 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
301 cccgccgcgc ttaatgcgcc gctacagggc gcgtccattc gccattcagg ctgcgcaact
361 gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat
421 gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa
481 cgacggccag tgaattgtaa tacgactcac tatagggcga attcgagctc ggtacccggg
541 gatcccggca tcagagcaga ttgtactgag agtgcaccat aatcggcatt ttcttttgcg
601 tttttatttg ttaactgtta attgtccttg ttcaaggatg ctgtctttga caacagatgt
661 tttcttgcct ttgatgttca gcaggaagct aggcgcaaac gttgattgtt tgtctgcgta
721 gaatcctctg tttgtcatat agcttgtaat cacgacattg tttcctttcg cttgaggtac
781 agcgaagtgt gagtaagtaa aggttacatc gttaggatca agatccattt ttaacacaag
841 gccagttttg ttcagcggct tgtatgggcc agttaaagaa ttagaaacat aaccaagcat
901 gtaaatatcg ttagacgtaa tgccgtcaat cgtcattttt gatccgcggg agtcagtgaa
961 cagataccat ttgccgttca ttttaaagac gttcgcgcgt tcaatttcat ctgttactgt
1021 gttagatgca atcagcggtt tcatcacttt tttcagtgtg taatcatcgt ttagctcaat
1081 cataccgaga gcgccgtttg ctaactcagc cgtgcgtttt ttatcgcttt gcagaagttt
1141 ttgactttct tgacggaaga atgatgtgct tttgccatag tatgctttgt taaataaaga
1201 ttcttcgcct tggtagccat cttcagttcc agtgtttgct tcaaatacta agtatttgtg
1261 gcctttatct tctacgtagt gaggatctct cagcgtatgg ttgtcgcctg agctgtagtt
1321 gccttcatcg atgaactgct gtacattttg atacgttttt ccgtcaccgt caaagattga
1381 tttataatcc tctacaccgt tgatgttcaa agagctgtct gatgctgata cgttaacttg
1441 tgcagttgtc agtgtttgtt tgccgtaatg tttaccggag aaatcagtgt agaataaacg
1501 gatttttccg tcagatgtaa atgtggctga acctgaccat tcttgtgttt ggtcttttag
1561 gatagaatca tttgcatcga atttgtcgct gtctttaaag acgcggccag cgtttttcca
1621 gctgtcaata gaagtttcgc cgactttttg atagaacatg taaatcgatg tgtcatccgc
1681 atttttagga tctccggcta atgcaaagac gatgtggtag ccgtgatagt ttgcgacagt
1741 gccgtcagcg ttttgtaatg gccagctgtc ccaaacgtcc aggccttttg cagaagagat
1801 atttttaatt gtggacgaat cgaactcagg aacttgatat ttttcatttt tttgctgttc
1861 agggatttgc agcatatcat ggcgtgtaat atgggaaatg ccgtatgttt ccttatatgg
1921 cttttggttc gtttctttcg caaacgcttg agttgcgcct cctgccagca gtgcggtagt
1981 aaaggttaat actgttgctt gttttgcaaa ctttttgatg ttcatcgttc atgtctcctt
2041 ttttatgtac tgtgttagcg gtctgcttct tccagccctc ctgtttgaag atggcaagtt
2101 agttacgcac aataaaaaaa gacctaaaat atgtaagggg tgacgccaaa gtatacactt
2161 tgccctttac acattttagg tcttgcctgc tttatcagta acaaacccgc gcgatttact
2221 tttcgacctc attctattag actctcgttt ggattgcaac tggtctattt tcctcttttg
2281 tttgatagaa aatcataaaa ggatttgcag actacgggcc taaagaacta aaaaatctat
2341 ctgtttcttt tcattctctg tattttttat agtttctgtt gcatgggcat aaagttgcgt
2401 cgacgccttg ctcaaattgg aatcaggttt gtgccaatac cagtagaaac agacgaagaa
2461 tccatggcgg ccgctgtcat gggcccttga caattaatca tccggctcgt ataatgtgtg
2521 gcgagaccat tggtctcagt tttagagcta gaaatagcaa gttaaaataa ggctagtccg
2581 ttatcaactt gaaaaagtgg caccgagtcg gtgctttttt tgagatctgt ccatacccat
2641 ggtctagaat gctcgagact tttcatactc ccgccattca gaagcttgag tattctatag
2701 tgtcacctaa atagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
2761 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
2821 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
2881 aacctgtcgt gccagaattc ttgaagacga aagggcctcg tgatacgcct atttttatag
2941 gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg
3001 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
3061 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
3121 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
3181 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
3241 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
3301 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg
3361 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
3421 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
3481 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
3541 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
3601 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgc agcaatggca
3661 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
3721 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
3781 ggctggttta ttgctgataa atctggagcc ggtgagcgtg gatctcgcgg tatcattgca
3841 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
3901 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
3961 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
4021 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
4081 cgtgagtttt cgttccactg agcgtcagac cccaattaca cgccactggc tgtgcttgct
4141 ggggtgacgg tggcaacggt ggcggccttg ctgggctatc gcgttggaaa gaaacgaggg
4201 aaaggggact gataaaccgg tcttagcccc tccccttggt gtccaaccgc tctgtaggcc
4261 tctcaggcgc cgctggtgcc gctggttgga cgccaagggt gaatccgcct cgataccctg
4321 attactcgct tcctgcgccc tctcaggcgg cgatagggga ctggtaaaac ggggattgcc
4381 cagacgcctc ccccgcccct tcaggggcac aaatgcggcc ccaacggggc cacgtagtgg
4441 tgcgtttttt gcgtttccac ccttttcttc cttttccctt ttaaaccttt taggacgtct
4501 acaggccacg taatccgtgg cctgtagagt ttaaaaaggg acggatttgt tgccattaag
4561 ggacggattt gttgttaaga agggacggat ttgttgttgt aaagggacgg atttgttgta
4621 ttgtgggacg cagatacagt gtccccttat acacaaggaa tgtcgaacgt ggcctcaccc
4681 ccaatggttt acaaaagcaa tgccctggtc gaggccgcgt atcgcctcag tgttcaggaa
4741 cagcggatcg ttctggcctg tattagccag gtgaagagga gcgagcctgt caccgatgaa
4801 gtgatgtatt cagtgacggc ggaggacata gcgacgatgg cgggtgtccc tatcgaatct
4861 tcctacaacc agctcaaaga agcggccctg cgcctgaaac ggcgggaagt ccggttaacc
4921 caagagccca atggcaaggg gaaaagaccg agtgtgatga ttaccggctg ggtgcaaaca
4981 atcatctacc gggagggtga gggccgtgta gaactcaggt tcaccaaaga catgctgccg
5041 tacctgacgg aactcaccaa acagttcacc aaatacgcct tggctgacgt ggccaagatg
5101 gacagcaccc acgcgatcag gctttacgag ctgctcatgc aatgggacag catcggccag
5161 cgcgaaatag aaattgacca gctgcgaaag tggtttcaac tggaaggccg gtatccctcg
5221 atcaaggact tcaagttgcg agtgcttgat ccagccgtga cgcagatcaa cgagcacagc
5281 ccgctacagg tggagtgggc gcagcgaaag accgggcgca aggtcacaca tctgttgttc
5341 agttttggac cgaagaagcc cgccaaggcg gtgggtaagg ccccagcgaa gcgcaaggcc
5401 gggaagattt cagatgctga gatcgcgaaa caggctcgcc ctggtgagac atgggaagcg
5461 gcccgcgctc gactaaccca gatgccgctt gcgcaaacta ttaactggcg aactacttac
5521 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
5581 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
5641 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
5701 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
5761 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
5821 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
5881 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
5941 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
6001 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
6061 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc
6121 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
6181 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
6241 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
6301 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
6361 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
6421 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
6481 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
6541 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
6601 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
6661 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
6721 agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
6781 cacagatgcg taaggagaaa ataccgcatc aggaaattgt aagcgttaat attttgttaa
6841 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca
6901 aaatccctta taaatcaaaa gaatagaccg agata
//
