| 质粒名称: | pDonor_ShCAST_kanR |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 127924 |
| 存储实验室: | Feng Zhang |
|
载体骨架基本信息 |
|
| 载体名称: | pUC19, R6K origin of replication |
| 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: | -- |
| 筛选标记: | -- |
| 载体抗性: | 卡那霉素,50 μg/mL |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 低拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | KanR, R6K origin of replication |
| 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: | pDonor_ShCAST_kanR was a gift from Feng Zhang (Addgene plasmid # 127924 ; http://n2t.net/addgene:127924 ; RRID:Addgene_127924) |
| 参考文献部分: | RNA-guided DNA insertion with CRISPR-associated transposases. Strecker J, Ladha A, Gardner Z, Schmid-Burgk JL, Makarova KS, Koonin EV, Zhang F. Science. 2019 Jul 5;365(6448):48-53. doi: 10.1126/science.aax9181. Epub 2019 Jun 6. 10.1126/science.aax9181 PubMed 31171706 |
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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 pDonor_ShCAST_kanR 3016 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Kanamycin resistance donor for ShCAST.
ACCESSION .
VERSION .
KEYWORDS pDonor_ShCAST_kanR
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3016)
AUTHORS Strecker J, Ladha A, Gardner Z, Schmid-Burgk JL, Makarova KS, Koonin
EV, Zhang F
TITLE RNA-guided DNA insertion with CRISPR-associated transposases.
JOURNAL Science. 2019 Jul 5;365(6448):48-53. doi: 10.1126/science.aax9181.
Epub 2019 Jun 6.
PUBMED 31171706
REFERENCE 2 (bases 1 to 3016)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..3016
/organism="synthetic DNA construct"
/mol_type="other DNA"
protein_bind complement(1..48)
/label=FRT
/bound_moiety="FLP recombinase from the Saccharomyces
cerevisiae 2u plasmid"
/note="FLP-mediated recombination occurs in the 8-bp core
sequence TCTAGAAA (Turan and Bode, 2011)."
CDS 435..1229
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(489..508)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 1099..1118
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
protein_bind complement(1337..1384)
/label=FRT
/bound_moiety="FLP recombinase from the Saccharomyces
cerevisiae 2u plasmid"
/note="FLP-mediated recombination occurs in the 8-bp core
sequence TCTAGAAA (Turan and Bode, 2011)."
terminator 1432..1518
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 1621..1715
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
rep_origin 2059..2415
/label=R6K gamma ori
/note="gamma replication origin from E. coli plasmid R6K;
requires the R6K initiator protein pi for replication"
oriT 2466..2574
/note="incP origin of transfer"
ORIGIN
1 gaagttccta tactttctag agaataggaa cttcggaata ggaacttcaa gatcccctga
61 ttccctttgt caacagcaat ggataattcg atttaacaaa tgcatggcgc aagggctgct
121 aaaggaagcg gaacacgtag aaagccagtc cgcagaaacg gtgctgaccc cggatgaatg
181 tcagctactg ggctatctgg acaagggaaa acgcaagcgc aaagagaaag caggtagctt
241 gcagtgggct tacatggcga tagctagact gggcggtttt atggacagca agcgaaccgg
301 aattgccagc tggggcgccc tctggtaagg ttgggaagcc ctgcaaagta aactggatgg
361 ctttcttgcc gccaaggatc tgatggcgca ggggatcaag atctgatcaa gagacaggat
421 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
481 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
541 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
601 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
661 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
721 aagtgccggg gcaggatctc ctgtcatccc accttgctcc tgccgagaaa gtatccatca
781 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
841 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
901 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
961 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
1021 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
1081 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
1141 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
1201 tctatcgcct tcttgacgag ttcttctgaa ttgaaaaagg aagagtatga ggatccaaca
1261 tttccaatca ctagtgaatt atctagaatt attccattga gtaagttttt aagcacatca
1321 gcttcaaaag cgctctgaag ttcctatact ttctagagaa taggaacttc ggaataggta
1381 cttcaagatc cccaattcga gatcgtccgg gccgcaagct cctagcggcg gatttgtcct
1441 actcaggaga gcgttcaccg acaaacaaca gataaaacga aaggcccagt ctttcgactg
1501 agcctttcgt tttatttgat gcctcaagct agagagtcat taccccaggc gtttaagggc
1561 accaataact gccttaaaaa aattacgccc cgccctgcca ctcatcgcag tctagcttgg
1621 attctcacca ataaaaaacg cccggcggca accgagcgtt ctgaacaaat ccagatggag
1681 ttctgaggtc attactggat ctatcaacag gagtccaagc tcagctaatt aaggcgacag
1741 tcaatttgtc attatgaaaa tacacaaaag ctttttccta tcttgcaaag cgacagctaa
1801 tttgtcacaa tcacggacaa cgacatctat tttgtcactg caaagaggtt atgctaaaac
1861 tgccaaagcg ctataatcta tactgtataa ggattttact gatgacaata atttgtcaca
1921 acgacatata attagtcact gtacacgtag agacgtagca atgctacctc tctacaatgg
1981 ttttgtatgg tgcactctca gtacaatctt gtagcaccgc ctacatacct cgctctgcta
2041 atcctgttac cagccggttg tcagccgtta agtgttcctg tgtcactcaa aattgctttg
2101 agaggctcta agggcttctc agtgcgttac atccctggct tgttgtccac aaccgttaaa
2161 ccttaaaagc tttaaaagcc ttatatattc ttttttttct tataaaactt aaaaccttag
2221 aggctattta agttgctgat ttatattaat tttattgttc aaacatgaga gcttagtacg
2281 tgaaacatga gagcttagta cgttagccat gagagcttag tacgttagcc atgagggttt
2341 agttcgttaa acatgagagc ttagtacgtt aaacttgaga gcttagtacg tgaaacatga
2401 gagcttagta cgtactatca acaggttgaa ctgcccatgt tctttcctgc gttatcagag
2461 cttatcggcc agcctcgcag agcaggattc ccgttgagca ccgccaggtg cgaataaggg
2521 acagtgaaga aggaacaccc gctcgcgggt gggcctactt cacctatcct gcccggctga
2581 cgccgttgga tacaccaagg aaagtctaca cgaacccttt ggcaaaatcc tgtatatcgt
2641 gcgaaaaagg atggatatac cgaaaaaatc gctataatga ccccgaagca gggttatgca
2701 gcggaaagta aaaaattttt agtttattag acatctccac aaaaggcgta gtgtacagtg
2761 acaaattatc tgtcgtcggt gacagattaa tgtcattgtg actatttaat tgtcgtcgtg
2821 acccatcagc gttgcttaat taattgatga caaattaaat gtcatcaata taatatgctc
2881 tgcaattatt atacaaagca attaaaacaa gcggataaaa ggacttgctt tcaacccacc
2941 cctaagttta atagttactg agggggatcc actagtgagc tcatgcatga tctcgaatta
3001 gcttcaaaag cgctct
//
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