| 质粒名称: | pUPRT::DHFR-D |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 58528 |
| 存储实验室: | David Sibley |
|
载体骨架基本信息 |
|
| 载体名称: | pDEST R4-R3 Vector II |
| 载体出品公司: | Invitrogen |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 4555bp |
| 完整质粒大小: | 7635bp |
| 载体修饰: | -- |
| 载体类型: | -- |
| 筛选标记: | 新霉素（G418） |
| 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: | DHFR cassette |
| 物种来源: | -- |
| 插入大小: | 3162bp |
| 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: | AgeI(未破坏) |
| 3' 克隆位点: | NotI (未破坏) |
| 5' 测序引物 | CMV-F |
| 3' 测序引物 | SV40pA-R |
|
文献引用方法 |
|
| 材料和方法部分: | pUPRT::DHFR-D was a gift from David Sibley (Addgene plasmid # 58528 ; http://n2t.net/addgene:58528 ; RRID:Addgene_58528) |
| 参考文献部分: | Efficient Gene Disruption in Diverse Strains of Toxoplasma gondii Using CRISPR/CAS9. Shen B, Brown KM, Lee TD, Sibley LD. MBio. 2014 May 13;5(3). pii: e01114-14. doi: 10.1128/mBio.01114-14. 10.1128/mBio.01114-14 PubMed 24825012 |
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 pUPRT::DHFR-D 7635 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION replace TgUPRT with DHFR by homologous recombination.
ACCESSION .
VERSION .
KEYWORDS pUPRT::DHFR-D
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7635)
AUTHORS Shen B, Brown KM, Lee TD, Sibley LD
TITLE Efficient Gene Disruption in Diverse Strains of Toxoplasma gondii
Using CRISPR/CAS9.
JOURNAL MBio. 2014 May 13;5(3). pii: e01114-14. doi: 10.1128/mBio.01114-14.
PUBMED 24825012
REFERENCE 2 (bases 1 to 7635)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7635
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(21..38)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(21..37)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(30..52)
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 365..387
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(425..443)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 511..615
/gene="bla"
/label=AmpR promoter
CDS 616..1476
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGSDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(834..853)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 1647..2235
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 2136..2155
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 2362..2384
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 2381..2397
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 2381..2397
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
protein_bind 2417..2437
/gene="mutant version of attB"
/label=attB4
/bound_moiety="BP Clonase(TM)"
/note="core recombination site for the Gateway(R) BP
reaction"
protein_bind 3416..3440
/gene="mutant version of attB"
/label=attB1
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
protein_bind complement(6607..6631)
/gene="mutant version of attB"
/label=attB2
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
protein_bind complement(7628..13)
/gene="mutant version of attB"
/label=attB3
/bound_moiety="BP Clonase(TM)"
/note="core recombination site for the Gateway(R) BP
reaction"
ORIGIN
1 ttatacatag ttgataattc actggccgtc gttttacaac gtcgtgactg ggaaaaccct
61 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc
121 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc
181 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat atggtgcact
241 ctcagtgcaa tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc
301 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
361 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga
421 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag
481 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
541 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
601 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
661 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
721 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
781 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
841 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
901 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
961 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
1021 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
1081 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
1141 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
1201 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
1261 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
1321 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggt cagatggtaa gccctcccgt
1381 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
1441 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
1501 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
1561 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
1621 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
1681 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
1741 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
1801 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
1861 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
1921 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
1981 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
2041 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
2101 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
2161 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
2221 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggt cttttgctgg
2281 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
2341 gcctttgagt gagctgatac cagcggataa caatttcaca caggaaacag ctatgaccat
2401 gattacgcca agctatcaac tttgtataga aaagttgtcc cgctgtgcct agtatcgaaa
2461 gctgtagagt tgccaccctg atcttctccc ttccaactct ccgcttcgac acacccccca
2521 acgacttgtt gtgaagctct ggcccccctg tttcggttgt gacaggctct ccaggtgctc
2581 ttgcactctc cagtccagct ggtgtcatag aaccgttatc tctagagtgc cccgaaagag
2641 ccccctcctt ttattcgttg tttcccctgg gtacgagcta tcttgtggct ttctttgctt
2701 ggggatgacc cttgtcgtga ccttctatgc gttctcgatg agaaaccgtc gagtcttcgt
2761 tgcatgcagg ggtttctgcg cgaagatact ctcccacaga gccacagaaa gtggtagccg
2821 ctgttttcca acgcagcgta cgtacctaga gatccttcca ggaaacgctg agcctccgct
2881 gtgactgatt ttctgcacgt tggcacgtgg atatcgcgat cccctgtccc ctccactgca
2941 tgcagtgtgg cggtaagacg agggccatat cgatcccatg gcagctgctg tgtggggtgc
3001 cgccatacgt aggggtacga tttctggtcc tgcccaactt cccgcctggt tgattcgagc
3061 aacttacaag gacaaggcaa ttaaccatct tgctattttt cctgttttcg tcgtcatcgg
3121 ctgtcctagc tctcacggct acccatcgtt ttcccaagcg ctgtcaagtg atgccattgg
3181 ttacactgga caaaggccct gcaaatacgc gtgcctccat catcgatccc cctttatcta
3241 ctttctcgct ttccgttatc ccttctgtct gctttttcct ttctcggctc cacctcattc
3301 ccctcgttcc ttgtagcggc cccggcttct acgttccgcc tgcgattccc gtattggtca
3361 cgcatccaca aggcgtattc cttttttcgt cggacctgtc cacagggctt ctaaaacaag
3421 tttgtacaaa aaagcaggct tcaagcttcg ccaggctgta aatcccgtga gtcgtcctca
3481 caaatcatca agcaggtgtc ctcagggaga ctgcctgact gagttatgct aattcctttc
3541 tactttggcg tggtcacggg ggcgcgccgg atccttaatt aagtctagca tgtcattcga
3601 ttttcacccc ccgcgtagtt cctgtgtgtc attcgttgtc gagacaactc tgtcccgccc
3661 cggtgctgtt ccatatgcgt gactttcccg caattttttc agactttcag gaaagacagg
3721 ctccggaacg atctcgtcca tgactggtaa atccacgaca ccgcaatggc ccccagcacc
3781 tctatctctc gtgccagggg actaacgttg tatgcgtctg cgtcttgtct ttttgcattc
3841 gctttccaaa aaagagagcc atccgttccc ccgcacattc aacgccgcga gtgcggtttt
3901 tgtctttttt gagtggtagg acgcttttca tgcgcgaact acgtggacat taagttccat
3961 tctctttttc gacagcacga aaccttgcat tcaaacccgc ccgcggaaga tccgatcttg
4021 ctgctgttcg cagtcccagt agcgtcctgt cggccgcgcc gtctctgttg gtgggcagcc
4081 gctacacctg ttatctgact gccgtgcgcg aaaatgacgc catttttggg aaaatcgggg
4141 aacttcattc tttaaaagta tgcggaggtt tcctttttct tctgttcgtt tctttttctc
4201 gggtttgata accgtgttcg atgtaagcac tttccgtctc tcctccgtgc tttgttcgac
4261 atcgagacca ggtgtgcaga tccttcgctt gtcgatccgg agacgcgtgt ctcgtagaac
4321 cttttcattt taccacacgg cagtgcggag cactgctctg agtgcagcag ggacgggtga
4381 agtttcgctt tagtagtgcg tttctgctct acggggcgtt gtcgtgtctg ggaagatgca
4441 gaaaccggtg tgtctggtcg tcgcgatgac ccccaagagg ggcatcggca tcaacaacgg
4501 cctcccgtgg ccccacttga ccacagattt caaacacttt cgtcgtgtga caaaaacgac
4561 gcccgaagaa gccagtcgcc tgaacgggtg gcttcccagg aaatttgcaa agacgggcga
4621 ctctggactt ccctctccat cagtcggcaa gagattcaac gccgttgtca tgggacggaa
4681 aaactgggaa agcatgcctc gaaagtttag acccctcgtg gacagattga acatcgtcgt
4741 ttcctcttcc ctcaaagaag aagacattgc ggcggagaag cctcaagctg aaggccagca
4801 gcgcgtccga gtctgtgctt cactcccagc agctctcagc cttctggagg aagagtacaa
4861 ggattctgtc gaccagattt ttgtcgtggg aggagcggga ctgtacgagg cagcgctgtc
4921 tctgggcgtt gcctctcacc tgtacatcac gcgtgtagcc cgcgagtttc cgtgcgacgt
4981 tttcttccct gcgttccccg gagatgacat tctttcaaac aaatcaactg ctgcgcaggc
5041 tgcagctcct gccgagtctg tgttcgttcc cttttgtccg gagctcggaa gagagaagga
5101 caatgaagcg acgtatcgac ccatcttcat ttccaagacc ttctcagaca acggggtacc
5161 ctacgactcc gtggttctcg agaagagaag gaagactgac gacgcagcca ctgcggaacc
5221 gagcaacgca atgagctcct tgacgtccac gagggagaca actcccgtgc acgggttgca
5281 ggctccttct tcggccgcag ccattgcccc ggtgttggcg tggatggacg aagaagaccg
5341 gaaaaaacgc gagcaaaagg aactgattcg ggccgttccg catgttcact ttagaggcca
5401 tgaagaattc cagtaccttg atctcattgc cgacattatt aacaatggaa ggacaatgga
5461 tgaccgaacg ggcgttggtg tcatctccaa attcggctgc actatgcgct actcgctgga
5521 tcaggccttt ccacttctca ccacaaagcg tgtgttctgg aaaggggtcc tcgaagagtt
5581 gctgtggttc attcgcggcg acacgaacgc aaaccatctt tctgagaagg gcgtgaagat
5641 ctgggacaag aatgtgacac gcgagttcct cgattcgcgc aatctccccc accgagaggt
5701 cggagacatc ggcccgggct acggcttcca gtggagacac ttcggcgcgg catacaaaga
5761 catgcacaca gactacacag ggcagggcgt cgaccagctg aagaatgtga tccagatgct
5821 gagaacgaat ccaacagatc gtcgcatgct catgactgcc tggaatcctg cagcgctgga
5881 cgaaatggcg ctgccgcctt gtcacttgtt gtgccagttc tacgtgaacg accagaagga
5941 gctgtcgtgc atcatgtatc agcggtcgtg cgatgtcggc ctcggcgtcc ccttcaacat
6001 cgcttcctat tcgcttttga cgctcatggt tgcacacgtc tgcaacctaa aacctaagga
6061 gttcattcac ttcatgggga acacgcatgt ctacacgaac catgtcgagg ctttaaaaga
6121 gcagctgcgg agagaaccga gaccgttccc cattgtgaac atcctcaaca aggaacgcat
6181 caaggaaatc gacgatttca ccgccgagga ttttgaggtc gtgggctacg tcccgcacgg
6241 acgaatccag atggagatgg ctgtctagcg gaaatacaga agctgcccgt ctctcgtttt
6301 cctctctttt cggagggatc agggagagtg cctcgggtcg gagagagctg acgagggggt
6361 gccagagacc cctgtgtcct ttatcgaaga aaagggatga ctcttcatgt ggcatttcac
6421 acagtctcac ctcgccttgt tttctttttg tcaatcagaa cgaaagcgag ttgcgggtga
6481 cgcagatgtg cgtgtatcca ctcgtgaatg cgttatcgtt ctgtatgccg ctagagtgct
6541 ggactgttgc tgtctgccca cgacagcaga caactttcct tctatgcact tgcaggatga
6601 attccgaccc agctttcttg tacaaagtgg tcgtctctag tttttttgac agaccgctga
6661 cggaatcgcg gactggaaaa agcacgcctc aagaggataa acaccatttt tatttttgtc
6721 tcgctcccac gtccctcacc atcggcagga acacagaaaa aaagttttag ttgtgaggct
6781 gcctggtgcg cgacaagcag ttgcgctttt ttgtgccacg gggtatatct ttggtcgcgc
6841 tcctgtcacg actgttgtta gtcatggacc ggacgatttc ttcgtatcag gcacagagtc
6901 ccgtacacct gtaacgggaa tgtttacgag gcggcgtgca tcccgttcct cgtccatctc
6961 gagttttatt tccctttcct ttgttcgaga ttctgctgtc cagaacgaga cagggaagcg
7021 aagtggtagt gccaggaagg gcgcagctta attgctcctc ttcaccttaa gtcgtttact
7081 tgtgcccaag aagtgttctt cacgattacc ttgtgtgttt ccgccgttct ttgctttttg
7141 ttcgcgctgc tcggaaacat ttacaacctc atatctagat gttgttgaag caatgtgaag
7201 aatggtccac gttacattct ctaacggaat tttaacggtg tgggatggac aagcaatgca
7261 tatgcgatgt cgaaccctcg aaaacccgta atgtgcgacc acgtcctggg tgaagcaaac
7321 gcgtttccac atacagtcgg tatcgatgtc acgcttgcca gttcgtgtag ggaagttggc
7381 gtgttttttt ttccacggtg gggattgaac agtgtcattg cccagaccat ggcagtacca
7441 agtctgctct gatgaagtgc gccaccagca ttctgttttt tcgctcccac gagggtggaa
7501 cctctagcac acaacttctc tcttctgagg ttgccgaaaa actgtctgct aatttatcga
7561 gttatttgaa ctacgggacc aacgggcaga gactgttttc actcggatgt cgtacagttg
7621 acgtcgacaa cttta
//
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