| 质粒名称: | pZHY501 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 36187 |
| 存储实验室: | Daniel Voytas |
|
载体骨架基本信息 |
|
| 载体名称: | pCP4 |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 7498 bp |
| 载体修饰: | 金门兼容（见Cermak等人，2011）TALENs表达载体。TAL-DNA结合域可通过BamHI和XbaI消化去除。 |
| 筛选标记: | LEU2 |
| 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: | Homodimeric FokI Nuclease |
| 基因突变: | delta152,+63 truncation to TAL backbone |
| 启动子 | TEV |
| 标签/融合蛋白: | ACV5（主干上的N终端） |
| 5' 测序引物 | ttggcgtcggcaaacagtgg |
| 3' 测序引物 | ttaaaagtttatctcaccg |
|
文献引用方法 |
|
| 材料和方法部分: | pZHY501 was a gift from Daniel Voytas (Addgene plasmid # 36187 ; http://n2t.net/addgene:36187 ; RRID:Addgene_36187) |
| 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 pZHY501 7498 bp ds-DNA circular SYN 13-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pZHY501
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7498)
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 7498)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7498
/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)"
promoter 2331..2735
/gene="S. cerevisiae LEU2"
/label=LEU2 promoter
CDS 2748..3842
/codon_start=1
/gene="S. cerevisiae LEU2"
/product="3-isopropylmalate dehydrogenase, required for
leucine biosynthesis"
/label=LEU2
/note="yeast auxotrophic marker"
/translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
IGGAAIDATGVPLPDEALEASKKVDAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
GLYEPCHGSAPDLPKNKVDPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
DLGGSNSTTEVGDAVAEEVKKILA"
misc_feature 4271..4774
/label=CEN/ARS
/note="S. cerevisiae CEN6 centromere fused to an
autonomously replicating sequence"
promoter 4829..4920
/gene="bla"
/label=AmpR promoter
rep_origin 5988..6576
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6477..6496
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 6730..6747
/label=L4440
/note="L4440 vector, forward primer"
terminator 6848..6942
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
promoter 6949..6967
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
promoter 7044..7442
/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 tatgcttctc cttacgcatc tgtgcggtat ttcacaccgc atatcgacgg tcgaggagaa
2101 cttctagtat atccacatac ctaatattat tgccttatta aaaatggaat cccaacaatt
2161 acatcaaaat ccacattctc ttcaaaatca attgtcctgt acttccttgt tcatgtgtgt
2221 tcaaaaacgt tatatttata ggataattat actctatttc tcaacaagta attggttgtt
2281 tggccgagcg gtctaaggcg cctgattcaa gaaatatctt gaccgcagtt aactgtggga
2341 atactcaggt atcgtaagat gcaagagttc gaatctctta gcaaccatta tttttttcct
2401 caacataacg agaacacaca ggggcgctat cgcacagaat caaattcgat gactggaaat
2461 tttttgttaa tttcagaggt cgcctgacgc atataccttt ttcaactgaa aaattgggag
2521 aaaaaggaaa ggtgagaggc cggaaccggc ttttcatata gaatagagaa gcgttcatga
2581 ctaaatgctt gcatcacaat acttgaagtt gacaatatta tttaaggacc tattgttttt
2641 tccaataggt ggttagcaat cgtcttactt tctaactttt cttacctttt acatttcagc
2701 aatatatata tatatttcaa ggatatacca ttctaatgtc tgcccctatg tctgccccta
2761 agaagatcgt cgttttgcca ggtgaccacg ttggtcaaga aatcacagcc gaagccatta
2821 aggttcttaa agctatttct gatgttcgtt ccaatgtcaa gttcgatttc gaaaatcatt
2881 taattggtgg tgctgctatc gatgctacag gtgtcccact tccagatgag gcgctggaag
2941 cctccaagaa ggttgatgcc gttttgttag gtgctgtggg tggtcctaaa tggggtaccg
3001 gtagtgttag acctgaacaa ggtttactaa aaatccgtaa agaacttcaa ttgtacgcca
3061 acttaagacc atgtaacttt gcatccgact ctcttttaga cttatctcca atcaagccac
3121 aatttgctaa aggtactgac ttcgttgttg tcagagaatt agtgggaggt atttactttg
3181 gtaagagaaa ggaagacgat ggtgatggtg tcgcttggga tagtgaacaa tacaccgttc
3241 cagaagtgca aagaatcaca agaatggccg ctttcatggc cctacaacat gagccaccat
3301 tgcctatttg gtccttggat aaagctaatg ttttggcctc ttcaagatta tggagaaaaa
3361 ctgtggagga aaccatcaag aacgaattcc ctacattgaa ggttcaacat caattgattg
3421 attctgccgc catgatccta gttaagaacc caacccacct aaatggtatt ataatcacca
3481 gcaacatgtt tggtgatatc atctccgatg aagcctccgt tatcccaggt tccttgggtt
3541 tgttgccatc tgcgtccttg gcctctttgc cagacaagaa caccgcattt ggtttgtacg
3601 aaccatgcca cggttctgct ccagatttgc caaagaataa ggttgaccct atcgccacta
3661 tcttgtctgc tgcaatgatg ttgaaattgt cattgaactt gcctgaagaa ggtaaggcca
3721 ttgaagatgc agttaaaaag gttttggatg caggtatcag aactggtgat ttaggtggtt
3781 ccaacagtac caccgaagtc ggtgatgctg tcgccgaaga agttaagaaa atccttgctt
3841 aaaaagattc tcttttttta tgatatttgt acataaactt tataaatgaa attcataata
3901 gaaacgacac gaaattacaa aatggaatat gttcataggg tagacgaaac tatatacgca
3961 atctacatac atttatcaag aaggagaaaa aggaggatag taaaggaata caggtaagca
4021 aattgatact aatggctcaa cgtgataagg aaaaagaatt gcactttaac attaatattg
4081 acaaggagga gggcaccaca caaaaagtta ggtgtaacag aaaatcatga aactacgatt
4141 cctaatttga tattggagga ttttctctaa aaaaaaaaaa atacaacaaa taaaaaacac
4201 tcaatgacct gaccatttga tgggctcacc aagctcttaa aacgggaatt tatggatctt
4261 aattaagacg gatcgcttgc ctgtaactta cacgcgcctc gtatctttta atgatggaat
4321 aatttgggaa tttactctgt gtttatttat ttttatgttt tgtatttgga ttttagaaag
4381 taaataaaga aggtagaaga gttacggaat gaagaaaaaa aaataaacaa aggtttaaaa
4441 aatttcaaca aaaagcgtac tttacatata tatttattag acaagaaaag cagattaaat
4501 agatatacat tcgattaacg ataagtaaaa tgtaaaatca caggattttc gtgtgtggtc
4561 ttctacacag acaagatgaa acaattcggc attaatacct gagagcagga agagcaagat
4621 aaaaggtagt atttgttggc gatcccccta gagtctttta catcttcgga aaacaaaaac
4681 tattttttct ttaatttctt tttttacttt ctatttttaa tttatatatt tatattaaaa
4741 aatttaaatt ataattattt ttatagcacg tgatgaaaag ccttagggtc aggtggcact
4801 tttcggggaa atgtgcgatg aacccctatt tgtttatttt tctaaataca ttcaaatatg
4861 tatccgctaa tgagacaata accctgataa atgcttcaat tatattgaaa aaggaagagt
4921 atgagtatcc aacatttccg tgtcgccctt attccctttt ttgcggcctt ctgccttcct
4981 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgcc
5041 cgagtgggtt acattgaact ggatctcaac agcggcaaaa tcttagagag ttttcgcccc
5101 gaagaacgct tcccaatgat gagcactttc aaagttctgc tatgtggcgc ggtgttatcc
5161 cgtattgatg ctgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
5221 gttgagtatt caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
5281 tgtagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatt
5341 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
5401 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
5461 cctgtagcaa tggcaacaac tctccgcaaa ctattaactg gcgaactact tactcttgct
5521 tcacgccagc aactcattga ctggatggag gcggacaaag ttgcaggacc acttctgcgc
5581 tcggcacttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
5641 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
5701 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
5761 tcactgatta agcattggta actgtcagac gaagggaaat aaatagtagc ccgcctgatg
5821 tgcgggcttt tttttggtgg tggttaccag tgtcgcttag ctaccgtagg tcctcttagg
5881 cggtccgatt agcgacatag tcatcaatgc cttgacggca aaatccctta acgtgagttt
5941 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
6001 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
6061 ttgccggatc aagagctact aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
6121 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
6181 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
6241 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
6301 ggctgaacgg ggggttcgtg ctcacagccc agcttggagc gaacgaccta caccgaactg
6361 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
6421 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
6481 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
6541 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
6601 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
6661 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
6721 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct
6781 ctccccgcgc gttggccgat tcattgatgc aggtcaacga catcacctgg ttgcgagcct
6841 tctgaaggac tcctgttgat agatccagta atgacctcag aactccatct ggatttgttc
6901 agaacgctcg gttgccgccg ggcgtttttt attggtgaga atcaggcgta atacgactca
6961 ctatagggcg aatggaactg gacgctcgcc tgtggtcgca gggtggctcg tacgaactta
7021 ccgcggagac atatcgataa tcacatagct tcaaaatgtt tctactcctt ttttactctt
7081 ccagattttc tcggactccg cgcatcgccg taccacttca aaacacccaa gcacagcata
7141 ctaaatttcc cctctttctt cctctagggt gtcgttaatt acccgtacta aaggtttgga
7201 aaagaaaaaa gagaccgcct cgtttctttt tcttcgtcga aaaaggcaat aaaaattttt
7261 atcacgtttc tttttcttga aaattttttt tttgattttt ttctctttcg atgacctccc
7321 attgatattt aagttaataa acggtcttca atttctcaag tttcagtttc atttttcttg
7381 ttctattaca acttttttta cttcttgctc attagaaaga aagcatagca atctaatcta
7441 agtttagatc taactcgaga aagatattgt atatatcgta acaataggag gttcaaca
//
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