| 质粒名称: | pCS2TAL3-RR |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 37276 |
| 存储实验室: | David Grunwald |
|
载体骨架基本信息 |
|
| 载体名称: | pCS2 |
| 载体抗性: | 氨苄霉素 |
| 空载体大小: | 4039 bp |
| 载体修饰: | The tal1c sequence from Dan Voytas lab's pTAL3 vector (1214-2210nt) consisting of a truncated N terminal and C terminal Tal protein regions and a LacZ screening module for use in Golden Gate cloning was PCR cloned into the KpnI and BamHI sites of the pCS2-Flag-GAAZFP-FokI-RR from Scot Wolfe's lab, making the new vector pCS2TAL3-RR. A Esp3I site in the FokI domain of the backbone vector was modified without changing the coding sequence. |
| 载体类型: | 哺乳细胞表达，Bacterial Expression |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 启动子 | CMV |
| 标签/融合蛋白: |
HA (N terminal on backbone)
FokI RR (C terminal on backbone)
N-terminal Tal from pTAL3 (N terminal on backbone)
C terminal Tal From pTAL3 (C terminal on backbone)
|
| 克隆方法: | 限制性内切酶 |
| 5' 测序引物 | TTGGCGTCGGCAAACAGTGG |
| 3' 测序引物 | GGCAACGCGATGGGACGT |
|
文献引用方法 |
|
| 材料和方法部分: | pCS2TAL3-RR was a gift from David Grunwald (爱迪基因 plasmid # 37276) |
| 参考文献部分: | Simple Methods for Generating and Detecting Locus-Specific Mutations Induced with TALENs in the Zebrafish Genome. Dahlem TJ, Hoshijima K, Jurynec MJ, Gunther D, Starker CG, Locke AS, Weis AM, Voytas DF, Grunwald DJ. PLoS Genet. 2012 Aug;8(8):e1002861. Epub 2012 Aug 16. 10.1371/journal.pgen.1002861 PubMed 22916025 |
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 pCS2TAL3-RR 5798 bp ds-DNA circular SYN 10-JAN-2018
DEFINITION TALEN backbone vector for use with the Golden Gate TALEN Kit in
place of pTAL1-4. Shorter N- and C-term tal protein segments
increases mutation induction; Fok I RR (improved version =
pCS2TAL3-RRR).
ACCESSION .
VERSION .
KEYWORDS pCS2TAL3-RR
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5798)
AUTHORS Dahlem TJ, Hoshijima K, Jurynec MJ, Gunther D, Starker CG, Locke AS,
Weis AM, Voytas DF, Grunwald DJ
TITLE Simple Methods for Generating and Detecting Locus-Specific Mutations
Induced with TALENs in the Zebrafish Genome.
JOURNAL PLoS Genet. 2012 Aug;8(8):e1002861. Epub 2012 Aug 16.
PUBMED 22916025
COMMENT This file is created by Vector NTI
http://www.biofeng.com/
COMMENT ORIGDB|GenBank
COMMENT VNTAUTHORNAME|biofeng.com|
COMMENT VNTNAME|pCS2TAL3-RR|
FEATURES Location/Qualifiers
source 1..5798
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 35..53
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind 35..52
/label=SP6
/note="SP6 promoter, forward primer"
regulatory 85..94
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 97..117
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 118..207
/codon_start=1
/product="three tandem HA epitope tags"
/label=3xHA
/translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
CDS complement(520..927)
/codon_start=1
/gene="lacZ fragment"
/product="LacZ-alpha fragment of beta-galactosidase"
/label=lacZ-alpha
/translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGARLQGG
TRQCIARCMHCLHCGHAATFCNLHKLACVQL"
primer_bind 777..799
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 791..808
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 792..808
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 818..837
/label=T7
/note="T7 promoter, forward primer"
promoter 818..836
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
promoter complement(884..902)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(923..939)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 947..963
/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(971..1001)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 1016..1037
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
CDS 1223..1810
/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
MEFFMKVYGYRGKHLGGSRKPDGAIYTVGSPIDYGVIVDTKAYSGGYNLPIGQAREMQR
YVEENQTRNKHINPNEWWKVYPSSVTEFKFLFVSGHFKGNYKAQLTRLNHITNCNGAVL
SVEELLIGGEMIKAGTLTLEEVRRKFNNGEINF"
promoter complement(1821..1838)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
polyA_signal 1843..1977
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(1867..1886)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 1921..1940
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
promoter complement(2094..2112)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(2133..2149)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 2157..2173
/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(2181..2211)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 2226..2247
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(2364..2381)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(2535..3123)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(2615..2634)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(3294..4154)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 3917..3936
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(4155..4259)
/gene="bla"
/label=AmpR promoter
rep_origin complement(4285..4740)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(4422..4443)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 4634..4653
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 4814..5798
/label=CMV IE94 promoter
/note="enhancer/promoter region of simian cytomegalovirus
major immediate early transcription unit IE94"
ORIGIN
1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
61 ctacttgttc tttttgcagg atctgccacc atggctccaa agaagaagcg taaggtatac
121 ccatacgatg ttcctgacta tgcgggctat ccctatgacg tcccggacta tgcaggatcg
181 tatccatatg acgttccaga ttacgctgct catggtaccg tggatctacg cacgctcggc
241 tacagtcagc agcagcaaga gaagatcaaa ccgaaggtgc gttcgacagt ggcgcagcac
301 cacgaggcac tggtgggcca tgggtttaca cacgcgcaca tcgttgcgct cagccaacac
361 ccggcagcgt tagggaccgt cgctgtcacg tatcagcaca taatcacggc gttgccagag
421 gcgacacacg aagacatcgt tggcgtcggc aaacagtggt ccggcgcacg cgccctggag
481 gccttgctca cggatgcggg ggagttgaga ggtccgccgt tacagttgga cacaggccaa
541 cttgtgaaga ttgcaaaacg tggcggcgtg accgcaatgg aggcagtgca tgcatcgcgc
601 aatgcactga cgggtgcccc cctggagacg ggcgccgcta cagggcgcgt cccattcgcc
661 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
721 gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca
781 gtcacgacgt tgtaaaacga cggccagtga gcgcgcgtaa tacgactcac tatagggcga
841 attgggtacc gggccccccc tcgaggtcct ccagcttttg ttccctttag tgagggttaa
901 ttgcgcgctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
961 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
1021 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccaccgg tcgtctccaa
1081 cgaccacctc gtcgccttgg cctgcctcgg cggacgtcct gccatggatg cagtgaaaaa
1141 gggattgccg cacgcgccgg aattgatcag aagagtcaat cgccgtattg gcgaacgcac
1201 gtcccatcgc gttgccggat cccaactagt caaaagtgaa ctggaggaga agaaatctga
1261 acttcgtcat aaattgaaat atgtgcctca tgaatatatt gaattaattg aaattgccag
1321 aaattccact caggatagaa ttcttgaaat gaaggtaatg gaatttttta tgaaagttta
1381 tggatataga ggtaaacatt tgggtggatc aaggaaaccg gacggagcaa tttatactgt
1441 cggatctcct attgattacg gtgtgatcgt ggatactaaa gcttatagcg gaggttataa
1501 tctgccaatt ggccaagcac gtgaaatgca acgatatgtc gaagaaaatc aaacacgaaa
1561 caaacatatc aaccctaatg aatggtggaa agtctatcca tcttctgtaa cggaatttaa
1621 gtttttattt gtgagtggtc actttaaagg aaactacaaa gctcagctta cacgattaaa
1681 tcatatcact aattgtaatg gagctgttct tagtgtagaa gagcttttaa ttggtggaga
1741 aatgattaaa gccggcacat taaccttaga ggaagtgcgc cggaaattta ataacggcga
1801 gataaacttt taatctagaa ctatagtgag tcgtattacg tagatccaga catgataaga
1861 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt
1921 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac
1981 aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa
2041 ttcgcggccg cggcgccaat gcattgggcc cggtacgtac ccagcttttg ttccctttag
2101 tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
2161 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
2221 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
2281 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
2341 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
2401 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
2461 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
2521 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
2581 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
2641 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
2701 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
2761 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
2821 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
2881 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
2941 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
3001 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
3061 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
3121 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
3181 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
3241 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
3301 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
3361 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
3421 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
3481 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
3541 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
3601 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
3661 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
3721 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
3781 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
3841 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
3901 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
3961 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
4021 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
4081 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
4141 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
4201 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
4261 acatttcccc gaaaagtgcc acctaaattg taagcgttaa tattttgtta aaattcgcgt
4321 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
4381 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
4441 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
4501 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
4561 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
4621 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
4681 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
4741 cccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
4801 tattacgcca gtcgaccata gccaattcaa tatggcgtat atggactcat gccaattcaa
4861 tatggtggat ctggacctgt gccaattcaa tatggcgtat atggactcgt gccaattcaa
4921 tatggtggat ctggacccca gccaattcaa tatggcggac ttggcaccat gccaattcaa
4981 tatggcggac ttggcactgt gccaactggg gaggggtcta cttggcacgg tgccaagttt
5041 gaggaggggt cttggccctg tgccaagtcc gccatattga attggcatgg tgccaataat
5101 ggcggccata ttggctatat gccaggatca atatataggc aatatccaat atggccctat
5161 gccaatatgg ctattggcca ggttcaatac tatgtattgg ccctatgcca tatagtattc
5221 catatatggg ttttcctatt gacgtagata gcccctccca atgggcggtc ccatatacca
5281 tatatggggc ttcctaatac cgcccatagc cactccccca ttgacgtcaa tggtctctat
5341 atatggtctt tcctattgac gtcatatggg cggtcctatt gacgtatatg gcgcctcccc
5401 cattgacgtc aattacggta aatggcccgc ctggctcaat gcccattgac gtcaatagga
5461 ccacccacca ttgacgtcaa tgggatggct cattgcccat tcatatccgt tctcacgccc
5521 cctattgacg tcaatgacgg taaatggccc acttggcagt acatcaatat ctattaatag
5581 taacttggca agtacattac tattggaagg acgccagggt acattggcag tactcccatt
5641 gacgtcaatg gcggtaaatg gcccgcgatg gctgccaagt acatccccat tgacgtcaat
5701 ggggaggggc aatgacgcaa atgggcgttc cattgacgta aatgggcggt aggcgtgcct
5761 aatgggaggt ctatataagc aatgctcgtt tagggaac
//
