| 质粒名称: | pSEVA-gRic6T |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 106401 |
| 存储实验室: | Sheng Yang |
|
载体骨架基本信息 |
|
| 载体名称: | pSEVA644 pTargetF |
| 完整质粒大小: | 4500 bp |
| 载体类型: | 细菌表达 |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 未知 |
|
插入基因信息 |
|
| 插入基因名称: | sgRNA |
| 物种来源: | Synthetic |
| 插入大小: | 96 bp |
| 启动子: | pj23119 |
| 克隆方法: | 未知 |
| 5’测序引物 | 未知 |
|
文献引用方法 |
|
| 材料和方法部分: | pSEVA-gRic6T was a gift from Sheng Yang (Addgene plasmid # 106401 ; http://n2t.net/addgene:106401 ; RRID:Addgene_106401) |
| 参考文献部分: | Genome editing and transcriptional repression in Pseudomonas putida KT2440 via the type II CRISPR system. Sun J, Wang Q, Jiang Y, Wen Z, Yang L, Wu J, Yang S. Microb Cell Fact. 2018 Mar 13;17(1):41. doi: 10.1186/s12934-018-0887-x. 10.1186/s12934-018-0887-x PubMed 29534717 |
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 pSEVA-gRic6T 4521 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION E.coli-Pseudomonas putida KT2440 shuttle plasmid containing sgRNA
targeting the KT2440 genome, and homologous arms..
ACCESSION .
VERSION .
KEYWORDS pSEVA-gRic6T
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4521)
AUTHORS Sun J, Wang Q, Jiang Y, Wen Z, Yang L, Wu J, Yang S
TITLE Genome editing and transcriptional repression in Pseudomonas putida
KT2440 via the type II CRISPR system.
JOURNAL Microb Cell Fact. 2018 Mar 13;17(1):41. doi:
10.1186/s12934-018-0887-x.
PUBMED 29534717
REFERENCE 2 (bases 1 to 4521)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..4521
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 25..59
/label=J23119 promoter
/note="bacterial promoter (Registry of Standard Biological
Parts BBa_J23119)"
misc_RNA 80..155
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
regulatory 233..242
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
terminator 1247..1341
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
promoter 1372..1400
/gene="intI1 (promoter lies within the coding sequence)"
/label=Pc promoter
/note="class 1 integron promoter"
oriT 2314..2422
/note="incP origin of transfer"
rep_origin 2516..3104
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 3005..3024
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(3158..3991)
/codon_start=1
/product="replication protein for the broad-host-range
plasmid pRO1600 from Pseudomonas aeruginosa "
/label=pRO1600 Rep
/translation="MASPPMVYKSNALVEAAYRLSVQEQRIVLACISQVKRSEPVTDEV
MYSVTAEDIATMAGVPIESSYNQLKEAALRLKRREVRLTQEPNGKGKRPSVMITGWVQT
IIYREGEGRVELRFTKDMLPYLTELTKQFTKYALADVAKMDSTHAIRLYELLMQWDSIG
QREIEIDQLRKWFQLEGRYPSIKDFKLRVLDPAVTQINEHSPLQVEWAQRKTGRKVTHL
LFSFGPKKPAKAVGKAPAKRKAGKISDAEIAKQARPGETWEAARARLTQMPLDLA"
rep_origin 4005..4356
/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)"
terminator 4429..4515
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
ORIGIN
1 ttaattaatt gacaccatga attcttgaca gctagctcag tcctaggtat aatgctagca
61 ggcagaaaat cgcaatcgtg ttttagagct agaaatagca agttaaaata aggctagtcc
121 gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt ttgaggagct cggtacccgg
181 ggatccggca gctctaccgc atacaccttg gccttcaaac gctgggcggc ctccatggcg
241 gcgttgaccg tgcgcgggtc ggagtagtgg ctcttgagca cggccacgct ctgcgtttcg
301 tccactcgag acagcttcag tacgtgctcg aacatctggg tcagttgtgc attgctcacc
361 ggcatcgggg ttctcctggg tggcgttgac aggggcgcac catggcagtg cctggtgctg
421 catctcggtg caaaataata gtgtgtacac cctacttgtg cagatgaaaa cgcatcctgc
481 cagcgagcgc aagtgatctg ttcaaacgtt acctatccac acattgacca gtatttacgg
541 taacaatcaa ccggatttcg tttcgatcag ctgtttgaaa aatattttca gctggctctt
601 ttcaaaggtc atttgtagcg tatacgctct aaatcgtcag gcggttgagc cgctggccac
661 ctatcagaac aagaggagac atacccgcac cttcgtcgct ggtggtaacg tcagcgccaa
721 tggcatccgc cagcattacc tgcgctatgg cggcaagggc catgcgctga tcctggtgcc
781 gggcatcacc agcccagcga tcacctgggg ctttgtcgcc gagcgccttg ggcattattt
841 cgacacctat gtgctggatg tgcgcgggcg tggcttgtct tcaagcggcc ccgacctgga
901 ctacggcacc gacgcttgtg ctgcggatat tcccgccttt gccgccgcgc tgggcctgga
961 tagctaccat ctgctcggtc actcgatggg ggcgcgtttt gccatccgcg cagccgccca
1021 aggggcaccg ggactgcagc ggctggtgct ggtcgaccca ccggtgtccg ggccgggccg
1081 ccgcgcctac ccgagcaagt tgccgtggta cgtcgattcg atccgccaag ccacggttgg
1141 catgagcggc gatgacatgc gcgccttctg tgccacctgg agcgacctgc aggcatgcaa
1201 gcttgcggcc gcgtcgtgac tgggaaaacc ctggcgacta gtcttggact cctgttgata
1261 gatccagtaa tgacctcaga actccatctg gatttgttca gaacgctcgg ttgccgccgg
1321 gcgtttttta ttggtgagaa tccaggggtc cccaataatt acgatttaaa tttgacataa
1381 gcctgttcgg ttcgtaaact gtaatgcaag tagcgtatgc gctcacgcaa ctggtccaga
1441 accttgaccg aacgcagcgg tggtaacggc gcagtggcgg ttttcatggc ttgttatgac
1501 tgtttttttg tacagcctat gcctcgggca tccaagcagc aagcgcgtta cgccgtgggt
1561 cgatgtttga tgttatggag cagcaacgat gttacgcagc agggcagtcg ccctaaaaca
1621 aagttaggtg gctcaagtat gggcatcatt cgcacatgta ggctcggccc tgaccaagtc
1681 aaatccatgc gggctgctct tgatcttttc ggtcgtgagt tcggagacgt agccacctac
1741 tcccaacatc agccggactc cgattacctc gggaacttgc tccgtagtaa gacattcatc
1801 gcgcttgctg ccttcgacca agaagcggtt gttggcgctc tcgcggctta cgttctgccc
1861 aagtttgagc agccgcgtag tgagatctat atctatgatc tcgcagtctc cggagagcac
1921 cggaggcagg gcattgccac cgcgctcatc aatctcctca agcatgaggc caacgcgctt
1981 ggtgcttatg tgatctacgt gcaagcagat tacggtgacg atcccgcagt ggctctctat
2041 acaaagttgg gcatacggga agaagtgatg cactttgata tcgacccaag taccgccacc
2101 taacaattcg ttcaagccga gatcgacttc ccggccgcgg agttgttcgg taaattggac
2161 aacggtccgc gcgttgtcct tttccgctgc ataaccctgc ttcggggtca ttatagcgat
2221 tttttcggta tatccatcct ttttcgcacg atatacagga ttttgccaaa gggttcgtgt
2281 agactttcct tggtgtatcc aacggcgtca gccgggcagg ataggtgaag taggcccacc
2341 cgcgagcggg tgttccttct tcactgtccc ttattcgcac ctggcggtgc tcaacgggaa
2401 tcctgctctg cgaggctggc cgtaggccgg ccgataatct catgaccaaa atcccttaac
2461 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
2521 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
2581 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
2641 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
2701 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
2761 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
2821 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
2881 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
2941 aggcggacag gcatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
3001 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
3061 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
3121 ccgtgaaagg caggccggtc cgtggtggcc acggcctcta ggccagatcc agcggcatct
3181 gggttagtcg agcgcgggcc gcttcccatg tctcaccagg gcgagcctgt ttcgcgatct
3241 cagcatctga aatcttcccg gccttgcgct tcgctggggc cttacccacc gccttggcgg
3301 gcttcttcgg tccaaaactg aacaacagat gtgtgacctt gcgcccggtc tttcgctgcg
3361 cccactccac ctgtagcggg ctgtgctcgt tgatctgcgt cacggctgga tcaagcactc
3421 gcaacttgaa gtccttgatc gagggatacc ggccttccag ttgaaaccac tttcgcagct
3481 ggtcaatttc tatttcgcgc tggccgatgc tgtcccattg catgagcagc tcgtaaagcc
3541 tgatcgcgtg ggtgctgtcc atcttggcca cgtcagccaa ggcgtatttg gtgaactgtt
3601 tggtgagttc cgtcaggtac ggcagcatgt ctttggtgaa cctgagttct acacggccct
3661 caccctcccg gtagatgatt gtttgcaccc agccggtaat catcacactc ggtcttttcc
3721 ccttgccatt gggctcttgg gttaaccgga cttcccgccg tttcaggcgc agggccgctt
3781 ctttgagctg gttgtaggaa gattcgatag ggacacccgc catcgtcgct atgtcctccg
3841 ccgtcactga atacatcact tcatcggtga caggctcgct cctcttcacc tggctaatac
3901 aggccagaac gatccgctgt tcctgaacac tgaggcgata cgcggcctcg accagggcat
3961 tgcttttgta aaccattggg ggtgaggcca cgttcgacat tccttgtgta taaggggaca
4021 ctgtatctgc gtcccacaat acaacaaatc cgtcccttta caacaacaaa tccgtccctt
4081 cttaacaaca aatccgtccc ttaatggcaa caaatccgtc cctttttaaa ctctacaggc
4141 cacggattac gtggcctgta gacgtcctaa aaggtttaaa agggaaaagg aagaaaaggg
4201 tggaaacgca aaaaacgcac cactacgtgg ccccgttggg gccgcatttg tgcccctgaa
4261 ggggcggggg aggcgtctgg gcaatccccg ttttaccagt cccctatcgc cgcctgagag
4321 ggcgcaggaa gcgagtaatc agggtatcga ggcggattca cccttggcgt ccaaccagcg
4381 gcaccagcgg cgcctgagag gggcgcgccc agctgtctag ggcggcggat ttgtcctact
4441 caggagagcg ttcaccgaca aacaacagat aaaacgaaag gcccagtctt tcgactgagc
4501 ctttcgtttt atttgatgcc t
//
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