| 质粒名称: | pDHt-SK |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 63615 |
| 存储实验室: | Seogchan Kang |
|
载体骨架基本信息 |
|
| 载体名称: | pDHt |
| 载体出品公司: | Mullins, E.D., Kang, S., 2001. Transformation: a tool for studying fungal pathogens of plants. Cell. Mol. Life Sci. 58, 2043–205 |
| 载体抗性: | 卡那霉素 |
| 空载体大小: | 7483 bp |
| 完整质粒大小: | -- |
| 载体修饰: | 用从pgreeni0000分离的0.8kb的HpaI-StuI片段替换含有MCS的pHT的300bp PvuII-PvuII片段 |
| 载体类型: | 真菌表达 |
| 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: | 限制性内切酶 |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | M13R |
| 3' 测序引物 | M13-F20 |
|
文献引用方法 |
|
| 材料和方法部分: | pDHt-SK was a gift from Seogchan Kang (Addgene plasmid # 63615 ; http://n2t.net/addgene:63615 ; RRID:Addgene_63615) |
| 参考文献部分: | A dual selection based, targeted gene replacement tool for Magnaporthe grisea and Fusarium oxysporum. Khang CH, Park SY, Lee YH, Kang S. Fungal Genet Biol. 2005 Jun;42(6):483-92. Epub 2005 Apr 25. 10.1016/j.fgb.2005.03.004 PubMed 15893252 |
买家导航
pDHtSK载体基本信息
pDHtSK载体图谱质粒图谱和多克隆位点信息
pDHtSK载体简介
pDHtSK载体序列
LOCUS pDHt-SK 7481 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION The multiple cloning site of pGreenII0000 cloned into binary vector
pDHt.
ACCESSION .
VERSION .
KEYWORDS pDHt-SK
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7481)
AUTHORS Khang CH, Park SY, Lee YH, Kang S
TITLE A dual selection based, targeted gene replacement tool for
Magnaporthe grisea and Fusarium oxysporum.
JOURNAL Fungal Genet Biol. 2005 Jun;42(6):483-92. Epub 2005 Apr 25.
PUBMED 15893252
REFERENCE 2 (bases 1 to 7481)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7481
/organism="synthetic DNA construct"
/mol_type="other DNA"
misc_feature 1..25
/label=LB T-DNA repeat
/note="left border repeat from nopaline C58 T-DNA"
polyA_signal 103..277
/label=CaMV poly(A) signal
/note="cauliflower mosaic virus polyadenylation signal"
protein_bind 585..606
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 621..651
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 659..675
/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)."
primer_bind 664..686
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 683..699
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 683..699
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
CDS 695..1060
/codon_start=1
/gene="lacZ fragment"
/product="LacZ-alpha fragment of beta-galactosidase"
/label=lacZ-alpha
/translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLSI
PSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
TDRPSQQLRSLNGEWKL"
primer_bind 718..738
/label=T3
/note="T3 promoter, forward primer"
promoter 720..738
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
misc_feature 751..858
/label=MCS
/note="pBluescript multiple cloning site"
primer_bind 775..791
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 775..791
/label=pBluescriptSK
/note="For pBluescript vector"
primer_bind complement(825..841)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(826..842)
/label=pBluescriptKS
/note="For pBluescript vector"
primer_bind complement(866..885)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(867..885)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(895..912)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(895..911)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(904..926)
/label=M13/pUC Forward
/note="In lacZ gene"
misc_feature 1204..1228
/label=RB T-DNA repeat
/note="right border repeat from nopaline C58 T-DNA"
CDS 2528..3157
/codon_start=1
/product="stability protein from the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
/label=pVS1 StaA
/translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
CDS 3591..4658
/codon_start=1
/product="replication protein from the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
/label=pVS1 RepA
/translation="GRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESWQA
AADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLSKR
DRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKPGR
VFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGEAL
ISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYRLA
RRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPILV
MRYRNLIEGEASAGS"
rep_origin 4724..4918
/label=pVS1 oriV
/note="origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
primer_bind complement(5154..5176)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
misc_feature 5262..5402
/label=bom
/note="basis of mobility region from pBR322"
primer_bind 5317..5336
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
rep_origin complement(5588..6176)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5668..5687)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(6263..7057)
/codon_start=1
/gene="aphA-3"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin"
/translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
ORIGIN
1 tggcaggata tattgtggtg taaacaaatt gacgcttaga caacttaata acacattgcg
61 gacgttttta atgtactgaa ttaacgccga attaattcgg gggatctgga ttttagtact
121 ggattttggt tttaggaatt agaaatttta ttgatagaag tattttacaa atacaaatac
181 atactaaggg tttcttatat gctcaacaca tgagcgaaac cctataggaa ccctaattcc
241 cttatctggg aactactcac acattattat ggagaaactc gattggcaag ctgctctagc
301 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc cttcatgttc
361 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
421 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
481 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
541 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
601 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
661 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagcgcgca
721 attaaccctc actaaaggga acaaaagctg gagctccacc gcggtggcgg ccgctctaga
781 actagtggat cccccgggct gcaggaattc gatatcaagc ttatcgatac cgtcgacctc
841 gagggggggc ccggtaccca attcgcccta tagtgagtcg tattacgcgc gctcactggc
901 cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc
961 agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc
1021 ccaacagttg cgcagcctga atggcgaatg gaaattgtaa gcgttaatgt tctggcgtaa
1081 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
1141 ctagagcagc ttgagcttgg atcagattgt cgtttcccgc cttcagttta aactatcagt
1201 gtttgacagg atatattggc gggtaaacct aagagaaaag agcgtttatt agaataacgg
1261 atatttaaaa gggcgtgaaa aggtttatcc gttcgtccat ttgtatgtgc atgccaacca
1321 cagggttccc ctcgggatca aagtactttg atccaacccc tccgctgcta tagtgcagtc
1381 ggcttctgac gttcagtgca gccgtcttct gaaaacgaca tgtcgcacaa gtcctaagtt
1441 acgcgacagg ctgccgccct gcccttttcc tggcgttttc ttgtcgcgtg ttttagtcgc
1501 ataaagtaga atacttgcga ctagaaccgg agacattacg ccatgaacaa gagcgccgcc
1561 gctggcctgc tgggctatgc ccgcgtcagc accgacgacc aggacttgac caaccaacgg
1621 gccgaactgc acgcggccgg ctgcaccaag ctgttttccg agaagatcac cggcaccagg
1681 cgcgaccgcc cggagctggc caggatgctt gaccacctac gccctggcga cgttgtgaca
1741 gtgaccaggc tagaccgcct ggcccgcagc acccgcgacc tactggacat tgccgagcgc
1801 atccaggagg ccggcgcggg cctgcgtagc ctggcagagc cgtgggccga caccaccacg
1861 ccggccggcc gcatggtgtt gaccgtgttc gccggcattg ccgagttcga gcgttcccta
1921 atcatcgacc gcacccggag cgggcgcgag gccgccaagg cccgaggcgt gaagtttggc
1981 ccccgcccta ccctcacccc ggcacagatc gcgcacgccc gcgagctgat cgaccaggaa
2041 ggccgcaccg tgaaagaggc ggctgcactg cttggcgtgc atcgctcgac cctgtaccgc
2101 gcacttgagc gcagcgagga agtgacgccc accgaggcca ggcggcgcgg tgccttccgt
2161 gaggacgcat tgaccgaggc cgacgccctg gcggccgccg agaatgaacg ccaagaggaa
2221 caagcatgaa accgcaccag gacggccagg acgaaccgtt tttcattacc gaagagatcg
2281 aggcggagat gatcgcggcc gggtacgtgt tcgagccgcc cgcgcacgtc tcaaccgtgc
2341 ggctgcatga aatcctggcc ggtttgtctg atgccaagct ggcggcctgg ccggccagct
2401 tggccgctga agaaaccgag cgccgccgtc taaaaaggtg atgtgtattt gagtaaaaca
2461 gcttgcgtca tgcggtcgct gcgtatatga tgcgatgagt aaataaacaa atacgcaagg
2521 ggaacgcatg aaggttatcg ctgtacttaa ccagaaaggc gggtcaggca agacgaccat
2581 cgcaacccat ctagcccgcg ccctgcaact cgccggggcc gatgttctgt tagtcgattc
2641 cgatccccag ggcagtgccc gcgattgggc ggccgtgcgg gaagatcaac cgctaaccgt
2701 tgtcggcatc gaccgcccga cgattgaccg cgacgtgaag gccatcggcc ggcgcgactt
2761 cgtagtgatc gacggagcgc cccaggcggc ggacttggct gtgtccgcga tcaaggcagc
2821 cgacttcgtg ctgattccgg tgcagccaag cccttacgac atatgggcca ccgccgacct
2881 ggtggagctg gttaagcagc gcattgaggt cacggatgga aggctacaag cggcctttgt
2941 cgtgtcgcgg gcgatcaaag gcacgcgcat cggcggtgag gttgccgagg cgctggccgg
3001 gtacgagctg cccattcttg agtcccgtat cacgcagcgc gtgagctacc caggcactgc
3061 cgccgccggc acaaccgttc ttgaatcaga acccgagggc gacgctgccc gcgaggtcca
3121 ggcgctggcc gctgaaatta aatcaaaact catttgagtt aatgaggtaa agagaaaatg
3181 agcaaaagca caaacacgct aagtgccggc cgtccgagcg cacgcagcag caaggctgca
3241 acgttggcca gcctggcaga cacgccagcc atgaagcggg tcaactttca gttgccggcg
3301 gaggatcaca ccaagctgaa gatgtacgcg gtacgccaag gcaagaccat taccgagctg
3361 ctatctgaat acatcgcgca gctaccagag taaatgagca aatgaataaa tgagtagatg
3421 aattttagcg gctaaaggag gcggcatgga aaatcaagaa caaccaggca ccgacgccgt
3481 ggaatgcccc atgtgtggag gaacgggcgg ttggccaggc gtaagcggct gggttgtctg
3541 ccggccctgc aatggcactg gaacccccaa gcccgaggaa tcggcgtgac ggtcgcaaac
3601 catccggccc ggtacaaatc ggcgcggcgc tgggtgatga cctggtggag aagttgaagg
3661 ccgcgcaggc cgcccagcgg caacgcatcg aggcagaagc acgccccggt gaatcgtggc
3721 aagcggccgc tgatcgaatc cgcaaagaat cccggcaacc gccggcagcc ggtgcgccgt
3781 cgattaggaa gccgcccaag ggcgacgagc aaccagattt tttcgttccg atgctctatg
3841 acgtgggcac ccgcgatagt cgcagcatca tggacgtggc cgttttccgt ctgtcgaagc
3901 gtgaccgacg agctggcgag gtgatccgct acgagcttcc agacgggcac gtagaggttt
3961 ccgcagggcc ggccggcatg gccagtgtgt gggattacga cctggtactg atggcggttt
4021 cccatctaac cgaatccatg aaccgatacc gggaagggaa gggagacaag cccggccgcg
4081 tgttccgtcc acacgttgcg gacgtactca agttctgccg gcgagccgat ggcggaaagc
4141 agaaagacga cctggtagaa acctgcattc ggttaaacac cacgcacgtt gccatgcagc
4201 gtacgaagaa ggccaagaac ggccgcctgg tgacggtatc cgagggtgaa gccttgatta
4261 gccgctacaa gatcgtaaag agcgaaaccg ggcggccgga gtacatcgag atcgagctag
4321 ctgattggat gtaccgcgag atcacagaag gcaagaaccc ggacgtgctg acggttcacc
4381 ccgattactt tttgatcgat cccggcatcg gccgttttct ctaccgcctg gcacgccgcg
4441 ccgcaggcaa ggcagaagcc agatggttgt tcaagacgat ctacgaacgc agtggcagcg
4501 ccggagagtt caagaagttc tgtttcaccg tgcgcaagct gatcgggtca aatgacctgc
4561 cggagtacga tttgaaggag gaggcggggc aggctggccc gatcctagtc atgcgctacc
4621 gcaacctgat cgagggcgaa gcatccgccg gttcctaatg tacggagcag atgctagggc
4681 aaattgccct agcaggggaa aaaggtcgaa aaggtctctt tcctgtggat agcacgtaca
4741 ttgggaaccc aaagccgtac attgggaacc ggaacccgta cattgggaac ccaaagccgt
4801 acattgggaa ccggtcacac atgtaagtga ctgatataaa agagaaaaaa ggcgattttt
4861 ccgcctaaaa ctctttaaaa cttattaaaa ctcttaaaac ccgcctggcc tgtgcataac
4921 tgtctggcca gcgcacagcc gaagagctgc aaaaagcgcc tacccttcgg tcgctgcgct
4981 ccctacgccc cgccgcttcg cgtcggccta tcgcggccgc tggccgctca aaaatggctg
5041 gcctacggcc aggcaatcta ccagggcgcg gacaagccgc gccgtcgcca ctcgaccgcc
5101 ggcgcccaca tcaaggcacc ctgcctcgcg cgtttcggtg atgacggtga aaacctctga
5161 cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa
5221 gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gcgcagccat gacccagtca
5281 cgtagcgata gcggagtgta tactggctta actatgcggc atcagagcag attgtactga
5341 gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca
5401 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
5461 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
5521 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
5581 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
5641 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
5701 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
5761 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
5821 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
5881 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
5941 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
6001 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
6061 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
6121 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
6181 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
6241 ttttggtcat gcattctagg tactaaaaca attcatccag taaaatataa tattttattt
6301 tctcccaatc aggcttgatc cccagtaagt caaaaaatag ctcgacatac tgttcttccc
6361 cgatatcctc cctgatcgac cggacgcaga aggcaatgtc ataccacttg tccgccctgc
6421 cgcttctccc aagatcaata aagccactta ctttgccatc tttcacaaag atgttgctgt
6481 ctcccaggtc gccgtgggaa aagacaagtt cctcttcggg cttttccgtc tttaaaaaat
6541 catacagctc gcgcggatct ttaaatggag tgtcttcttc ccagttttcg caatccacat
6601 cggccagatc gttattcagt aagtaatcca attcggctaa gcggctgtct aagctattcg
6661 tatagggaca atccgatatg tcgatggagt gaaagagcct gatgcactcc gcatacagct
6721 cgataatctt ttcagggctt tgttcatctt catactcttc cgagcaaagg acgccatcgg
6781 cctcactcat gagcagattg ctccagccat catgccgttc aaagtgcagg acctttggaa
6841 caggcagctt tccttccagc catagcatca tgtccttttc ccgttccaca tcataggtgg
6901 tccctttata ccggctgtcc gtcattttta aatataggtt ttcattttct cccaccagct
6961 tatatacctt agcaggagac attccttccg tatcttttac gcagcggtat ttttcgatca
7021 gttttttcaa ttccggtgat attctcattt tagccattta ttatttcctt cctcttttct
7081 acagtattta aagatacccc aagaagctaa ttataacaag acgaactcca attcactgtt
7141 ccttgcattc taaaacctta aataccagaa aacagctttt tcaaagttgt tttcaaagtt
7201 ggcgtataac atagtatcga cggagccgat tttgaaaccg cggtgatcac aggcagcaac
7261 gctctgtcat cgttacaatc aacatgctac cctccgcgag atcatccgtg tttcaaaccc
7321 ggcagcttag ttgccgttct tccgaatagc atcggtaaca tgagcaaagt ctgccgcctt
7381 acaacggctc tcccgctgac gccgtcccgg actgatgggc tgcctgtatc gagtggtgat
7441 tttgtgccga gctgccggtc ggggagctgt tggctggctg g
//
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