| 质粒名称: | pZHY500 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 36186 |
| 存储实验室: | Daniel Voytas |
|
载体骨架基本信息 |
|
| 载体名称: | pCP3 |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 6561 bp |
| 完整质粒大小: | -- |
| 载体修饰: | TALENs 的 Golden Gate 兼容（见 Cermak, et al., 2011）表达载体。TAL-DNA 结合域可以通过 BamHI 和 XbaI 消化去除。 |
| 载体类型: | 酵母表达 |
| 筛选标记: | HIS3 |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | FokI homodimeric Nuclease domain |
| 物种来源: | -- |
| 插入大小: | -- |
| 基因突变: | delta152,+63 truncation to TAL backbone |
| GenBank编号 | -- |
| 启动子 | TEV |
| 标签/融合蛋白: | ACV5（主干上的N终端） |
| 克隆方法: | -- |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | atcgcgcaatgcactgacgggtg |
| 3' 测序引物 | ttaaaagtttatctcaccg |
|
文献引用方法 |
|
| 材料和方法部分: | pZHY500 was a gift from Daniel Voytas (Addgene plasmid # 36186 ; http://n2t.net/addgene:36186 ; RRID:Addgene_36186) |
| 参考文献部分: | TALENs enable efficient plant genome engineering. Zhang Y, Zhang F, Li X, Baller JA, Qi Y, Starker CG, Bogdanove AJ, Voytas DF. Plant Physiol. 2012 Nov 2. 10.1104/pp.112.205179 PubMed 23124327 |
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 pZHY500 6561 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pZHY500
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6561)
AUTHORS Zhang Y, Zhang F, Li X, Baller JA, Qi Y, Starker CG, Bogdanove AJ,
Voytas DF
TITLE TALENs enable efficient plant genome engineering.
JOURNAL Plant Physiol. 2012 Nov 2.
PUBMED 23124327
REFERENCE 2 (bases 1 to 6561)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6561
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS 16..36
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 37..63
/codon_start=1
/product="epitope tag from the baculovirus Autographa
californica GP64 envelope fusion protein"
/label=AcV5 tag
/translation="SWKDASGWS"
CDS complement(370..777)
/codon_start=1
/gene="lacZ fragment"
/product="LacZ-alpha fragment of beta-galactosidase"
/label=lacZ-alpha
/translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGARLQGG
TRQCIARCMHCLHCGHAATFCNLHKLACVQL"
primer_bind 627..649
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 641..658
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 642..658
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 668..686
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(734..754)
/label=T3
/note="T3 promoter, forward primer"
promoter complement(734..752)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(773..789)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(773..789)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(786..808)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 797..813
/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(821..851)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 866..887
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
CDS 1073..1660
/codon_start=1
/product="nonspecific DNA cleavage domain of the FokI
endonuclease (Li et al., 1992)"
/label=FokI cleavage domain
/note="must dimerize to be catalytically active"
/translation="QLVKSELEEKKSELRHKLKYVPHEYIELIEIARNSTQDRILEMKV
MEFFMKVYGYRGKHLGGSRKPDGAIYTVGSPIDYGVIVDTKAYSGGYNLPIGQADEMQR
YVEENQTRNKHINPNEWWKVYPSSVTEFKFLFVSGHFKGNYKAQLTRLNHITNCNGAVL
SVEELLIGGEMIKAGTLTLEEVRRKFNNGEINF"
terminator 1683..1935
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
promoter complement(1977..1995)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
terminator 1997..2036
/label=fd terminator
/note="central terminator from bacteriophage fd (Otsuka and
Kunisawa, 1982)"
CDS complement(2336..2995)
/codon_start=1
/gene="S. cerevisiae HIS3"
/product="imidazoleglycerol-phosphate dehydratase, required
for histidine biosynthesis"
/label=HIS3
/note="yeast auxotrophic marker"
/translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
KEALLARGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHFL
ESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
promoter complement(2996..3183)
/gene="S. cerevisiae HIS3"
/label=HIS3 promoter
misc_feature 3334..3837
/label=CEN/ARS
/note="S. cerevisiae CEN6 centromere fused to an
autonomously replicating sequence"
promoter 3892..3983
/gene="bla"
/label=AmpR promoter
rep_origin 5051..5639
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5540..5559
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 5793..5810
/label=L4440
/note="L4440 vector, forward primer"
terminator 5911..6005
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
promoter 6012..6030
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
promoter 6107..6505
/gene="S. cerevisiae TEF1"
/label=TEF1 promoter
/note="promoter for EF-1-alpha"
ORIGIN
1 atggcttcct cccctccaaa gaaaaagaga aaggttagtt ggaaggacgc aagtggttgg
61 tctagaatgg tggatctacg cacgctcggc tacagtcagc agcagcaaga gaagatcaaa
121 ccgaaggtgc gttcgacagt ggcgcagcac cacgaggcac tggtgggcca tgggtttaca
181 cacgcgcaca tcgttgcgct cagccaacac ccggcagcgt tagggaccgt cgctgtcacg
241 tatcagcaca taatcacggc gttgccagag gcgacacacg aagacatcgt tggcgtcggc
301 aaacagtggt ccggcgcacg cgccctggag gccttgctca cggatgcggg ggagttgaga
361 ggtccgccgt tacagttgga cacaggccaa cttgtgaaga ttgcaaaacg tggcggcgtg
421 accgcaatgg aggcagtgca tgcatcgcgc aatgcactga cgggtgcccc cctggagacg
481 ggcgccgcta cagggcgcgt cccattcgcc attcaggctg cgcaactgtt gggaagggcg
541 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg
601 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga
661 gcgcgcgtaa tacgactcac tatagggcga attgggtacc gggccccccc tcgaggtcct
721 ccagcttttg ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc
781 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
841 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
901 cactgcccgc tttccaccgg tcgtctccaa cgaccacctc gtcgccttgg cctgcctcgg
961 cggacgtcct gccatggatg cagtgaaaaa gggattgccg cacgcgccgg aattgatcag
1021 aagagtcaat cgccgtattg gcgaacgcac gtcccatcgc gttgccggat cccagctagt
1081 gaaatctgaa ttggaagaga agaaatctga acttagacat aaattgaaat atgtgccaca
1141 tgaatatatt gaattgattg aaatcgcaag aaattcaact caggatagaa tccttgaaat
1201 gaaggtgatg gagttcttta tgaaggttta tggttatcgt ggtaaacatt tgggtggatc
1261 aaggaaacca gacggagcaa tttatactgt cggatctcct attgattacg gtgtgatcgt
1321 tgatactaag gcatattcag gaggttataa tcttccaatt ggtcaagcag atgaaatgca
1381 aagatatgtc gaagagaatc aaacaagaaa caagcatatc aaccctaatg aatggtggaa
1441 agtctatcca tcttcagtaa cagaatttaa gttcttgttt gtgagtggtc atttcaaagg
1501 aaactacaaa gctcagctta caagattgaa tcatatcact aattgtaatg gagctgttct
1561 tagtgtagaa gagcttttga ttggtggaga aatgattaaa gctggtacat tgacacttga
1621 ggaagtgaga aggaaattta ataacggtga gataaacttt taataggagc tcgaatttcc
1681 ccgatcgttc aaacatttgg caataaagtt tcttaagatt gaatcctgtt gccggtcttg
1741 cgatgattat catataattt ctgttgaatt acgttaagca tgtaataatt aacatgtaat
1801 gcatgacgtt atttatgaga tgggttttta tgattagagt cccgcaatta tacatttaat
1861 acgcgataga aaacaaaata tagcgcgcaa actaggataa attatcgcgc gcggtgtcat
1921 ctatgttact agatcgtggg cccacattcc tgtagaactc ggtgagactt ttgttccctt
1981 tagtgagggt taattgtcca aaaaaaaagg ctccaaaagg agcctttaat tgtttcggtt
2041 tatgcttagg acaccattga ttaacgctta caatttcctg atgcggtatt ttctccttac
2101 gcatctgtgc ggtatttcac accgcataga tccgtcgagt tcaagagaaa aaaaaagaaa
2161 aagcaaaaag aaaaaaggaa agcgcgcctc gttcagaatg acacgtatag aatgatgcat
2221 taccttgtca tcttcagtat catactgttc gtataaatac ttactgacat tcatagctat
2281 acatatatac acatgtatat atatcgtatg ctggagcttt aaataatcgg tgtcactaca
2341 taagaacacc tttggtggag ggaacatcgt tggttccatt gggcgaggtg gcttctctta
2401 tggcaaccgc aagagccttg aacgcactct cactacggtg atggtcattc ttgcctcgca
2461 gacaatcaac gtggagggta attctgcttg cctctgcaaa actttcaaga aaatgcggga
2521 tcatctcgca agagaggtct cctactttct ccctttgcaa accaagttcg acaactgcgt
2581 atggcctgtt agaaaggtct accaccgctc tggaaagtgc ctcatccaaa ggcgcaaatc
2641 ctgatccaaa cctttttact ccacgcgcca gtagagcctc tttaaaagcc tgaccgagag
2701 caatcccgca gtcttcagtg gtgtgatggt cgtctatgtg taagtcacca atgcactcaa
2761 cgattagcga ccagccggaa tgcttggcca gagcatgtat catgtggtcc agaaacccta
2821 tacctgtgtg gacgttaatc acttgcgatt gtgtggcctg ttctgctact gcttctgcct
2881 ctttttctgg gaagatcgag tgctctatcg ctaggggacc accctttaaa gagatagcaa
2941 tctgaatctt ggtttcattt gtaatacgct ttactagggc tttctgctct gtcatctttg
3001 ccttcgttta tcttgcctgc tcatttttta gtatattctt cgaagaaatc acattacttt
3061 atataatgta taattcatta tgtgataatg ccaatcgcta agaaaaaaaa agagtcatcc
3121 gctaggtgga aaaaaaaaaa tgaaaatcat taccgaggca taaaaaaata tagagtgtac
3181 tagaggaggc caagagtaat agaaaaagaa aattgcggga aaggactgtg ttatgacttc
3241 cctgactaat gccgtgttca aacgatacct ggcagtgact cctagcgctc accaagctct
3301 taaaacggga atttatggat cttaattaag acggatcgct tgcctgtaac ttacacgcgc
3361 ctcgtatctt ttaatgatgg aataatttgg gaatttactc tgtgtttatt tatttttatg
3421 ttttgtattt ggattttaga aagtaaataa agaaggtaga agagttacgg aatgaagaaa
3481 aaaaaataaa caaaggttta aaaaatttca acaaaaagcg tactttacat atatatttat
3541 tagacaagaa aagcagatta aatagatata cattcgatta acgataagta aaatgtaaaa
3601 tcacaggatt ttcgtgtgtg gtcttctaca cagacaagat gaaacaattc ggcattaata
3661 cctgagagca ggaagagcaa gataaaaggt agtatttgtt ggcgatcccc ctagagtctt
3721 ttacatcttc ggaaaacaaa aactattttt tctttaattt ctttttttac tttctatttt
3781 taatttatat atttatatta aaaaatttaa attataatta tttttatagc acgtgatgaa
3841 aagccttagg gtcaggtggc acttttcggg gaaatgtgcg atgaacccct atttgtttat
3901 ttttctaaat acattcaaat atgtatccgc taatgagaca ataaccctga taaatgcttc
3961 aattatattg aaaaaggaag agtatgagta tccaacattt ccgtgtcgcc cttattccct
4021 tttttgcggc cttctgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
4081 atgctgaaga tcagttgggt gcccgagtgg gttacattga actggatctc aacagcggca
4141 aaatcttaga gagttttcgc cccgaagaac gcttcccaat gatgagcact ttcaaagttc
4201 tgctatgtgg cgcggtgtta tcccgtattg atgctgggca agagcaactc ggtcgccgca
4261 tacactattc tcagaatgac ttggttgagt attcaccagt cacagaaaag catcttacgg
4321 atggcatgac agtaagagaa ttatgtagtg ctgccataac catgagtgat aacactgcgg
4381 ccaacttact tctgacaacg attggaggac cgaaggagct aaccgctttt ttgcacaaca
4441 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
4501 acgacgagcg tgacaccacg atgcctgtag caatggcaac aactctccgc aaactattaa
4561 ctggcgaact acttactctt gcttcacgcc agcaactcat tgactggatg gaggcggaca
4621 aagttgcagg accacttctg cgctcggcac ttccggctgg ctggtttatt gctgataaat
4681 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
4741 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
4801 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gacgaaggga
4861 aataaatagt agcccgcctg atgtgcgggc ttttttttgg tggtggttac cagtgtcgct
4921 tagctaccgt aggtcctctt aggcggtccg attagcgaca tagtcatcaa tgccttgacg
4981 gcaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
5041 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
5101 caccgctacc agcggtggtt tgtttgccgg atcaagagct actaactctt tttccgaagg
5161 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
5221 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
5281 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
5341 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgctcacag cccagcttgg
5401 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
5461 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
5521 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
5581 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
5641 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
5701 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
5761 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
5821 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattga tgcaggtcaa
5881 cgacatcacc tggttgcgag ccttctgaag gactcctgtt gatagatcca gtaatgacct
5941 cagaactcca tctggatttg ttcagaacgc tcggttgccg ccgggcgttt tttattggtg
6001 agaatcaggc gtaatacgac tcactatagg gcgaatggaa ctggacgctc gcctgtggtc
6061 gcagggtggc tcgtacgaac ttaccgcgga gacatatcga taatcacata gcttcaaaat
6121 gtttctactc cttttttact cttccagatt ttctcggact ccgcgcatcg ccgtaccact
6181 tcaaaacacc caagcacagc atactaaatt tcccctcttt cttcctctag ggtgtcgtta
6241 attacccgta ctaaaggttt ggaaaagaaa aaagagaccg cctcgtttct ttttcttcgt
6301 cgaaaaaggc aataaaaatt tttatcacgt ttctttttct tgaaaatttt ttttttgatt
6361 tttttctctt tcgatgacct cccattgata tttaagttaa taaacggtct tcaatttctc
6421 aagtttcagt ttcatttttc ttgttctatt acaacttttt ttacttcttg ctcattagaa
6481 agaaagcata gcaatctaat ctaagtttag atctaactcg agaaagatat tgtatatatc
6541 gtaacaatag gaggttcaac a
//
