| 质粒名称: | pTAL4 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 31035 |
| 存储实验室: | Adam Bogdanove, Daniel Voytas |
|
载体骨架基本信息 |
|
| 载体名称: | pDW1789_Leu |
| 载体抗性: | 氨苄青霉素（50 ug/ml） |
| 空载体大小: | 6519 bp |
| 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: | TALEN-FokI nuclease backbone |
| 物种来源: | Xanthamonas oryzae |
| 插入大小: | 1948 bp |
| 标签/融合蛋白: |
FokI nuclease（插入物上的C末端） AcV5 |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | BamHI（未销毁） |
| 3' 克隆位点: | BamHI（未销毁） |
| 5' 测序引物 | TEF_seq (ggtcttcaatttctcaagtttc) TAL_F1 (ttggcgtcggcaaacagtgg) |
| 3' 测序引物 | homodimer_rev (aattcagatttcactagctg) TAL_R2 (ggcgacgaggtggtcgttg |
|
文献引用方法 |
|
| 材料和方法部分: | pTAL4 was a gift from Adam Bogdanove & Daniel Voytas (Addgene plasmid # 31035 ; http://n2t.net/addgene:31035 ; RRID:Addgene_31035) |
| 参考文献部分: | Efficient design and assembly of custom TALEN and other TAL effector-based constructs for DNA targeting. Cermak T, Doyle EL, Christian M, Wang L, Zhang Y, Schmidt C, Baller JA, Somia NV, Bogdanove AJ, Voytas DF. Nucleic Acids Res. 2011 Apr 14. ():. 10.1093/nar/gkr218 PubMed 21493687 |
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 pTAL4 8467 bp ds-DNA circular SYN 13-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pTAL4
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8467)
AUTHORS Cermak T, Doyle EL, Christian M, Wang L, Zhang Y, Schmidt C, Baller
JA, Somia NV, Bogdanove AJ, Voytas DF
TITLE Efficient design and assembly of custom TALEN and other TAL
effector-based constructs for DNA targeting.
JOURNAL Nucleic Acids Res. 2011 Apr 14. ():.
PUBMED 21493687
REFERENCE 2 (bases 1 to 8467)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..8467
/organism="synthetic DNA construct"
/mol_type="other DNA"
terminator 35..129
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
promoter 136..154
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
promoter 231..629
/gene="S. cerevisiae TEF1"
/label=TEF1 promoter
/note="promoter for EF-1-alpha"
CDS 701..721
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 722..748
/codon_start=1
/product="epitope tag from the baculovirus Autographa
californica GP64 envelope fusion protein"
/label=AcV5 tag
/translation="SWKDASGWS"
CDS complement(1514..1921)
/codon_start=1
/gene="lacZ fragment"
/product="LacZ-alpha fragment of beta-galactosidase"
/label=lacZ-alpha
/translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGARLQGG
TRQCIARCMHCLHCGHAATFCNLHKLACVQL"
primer_bind 1771..1793
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 1785..1802
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 1786..1802
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 1812..1830
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(1878..1898)
/label=T3
/note="T3 promoter, forward primer"
promoter complement(1878..1896)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(1917..1933)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(1917..1933)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(1930..1952)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 1941..1957
/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(1965..1995)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 2010..2031
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
CDS 2727..3314
/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 3337..3589
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
promoter complement(3631..3649)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
terminator 3651..3690
/label=fd terminator
/note="central terminator from bacteriophage fd (Otsuka and
Kunisawa, 1982)"
promoter 3985..4389
/gene="S. cerevisiae LEU2"
/label=LEU2 promoter
CDS 4402..5496
/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 5925..6428
/label=CEN/ARS
/note="S. cerevisiae CEN6 centromere fused to an
autonomously replicating sequence"
promoter 6483..6574
/gene="bla"
/label=AmpR promoter
rep_origin 7642..8230
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 8131..8150
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 8384..8401
/label=L4440
/note="L4440 vector, forward primer"
ORIGIN
1 tcaacgacat cacctggttg cgagccttct gaaggactcc tgttgataga tccagtaatg
61 acctcagaac tccatctgga tttgttcaga acgctcggtt gccgccgggc gttttttatt
121 ggtgagaatc aggcgtaata cgactcacta tagggcgaat ggaactggac gctcgcctgt
181 ggtcgcaggg tggctcgtac gaacttaccg cggagacata tcgataatca catagcttca
241 aaatgtttct actccttttt tactcttcca gattttctcg gactccgcgc atcgccgtac
301 cacttcaaaa cacccaagca cagcatacta aatttcccct ctttcttcct ctagggtgtc
361 gttaattacc cgtactaaag gtttggaaaa gaaaaaagag accgcctcgt ttctttttct
421 tcgtcgaaaa aggcaataaa aatttttatc acgtttcttt ttcttgaaaa tttttttttt
481 gatttttttc tctttcgatg acctcccatt gatatttaag ttaataaacg gtcttcaatt
541 tctcaagttt cagtttcatt tttcttgttc tattacaact ttttttactt cttgctcatt
601 agaaagaaag catagcaatc taatctaagt ttagatctaa ctcgagaaag atattgtata
661 tatcgtaaca ataggaggtt caacaatggc ttcctcccct ccaaagaaaa agagaaaggt
721 tagttggaag gacgcaagtg gttggtctag aatgcatgcg gatcccattc gtccgcgcag
781 gccaagtcct gcccgcgagc ttctgcccgg accccaaccg gatagggttc agccgactgc
841 agatcgtggg gtgtctgcgc ctgctggcag ccctctggat ggcttgcccg ctcggcggac
901 ggtgtcccgg acccggctgc catctccccc tgcgccctca cctgcgttct cggcgggcag
961 cttcagcgat ctgctccgtc cgttcgatcc gtcgcttctt gatacatcgc ttcttgattc
1021 gatgcctgcc gtcggcacgc cgcatacagc ggctgcccca gcagagtggg atgaggcgca
1081 atcggctctg cgtgcagccg atgacccgcc acccaccgtg cgtgtcgctg tcactgccgc
1141 gcggccgccg cgcgccaagc cggccccgcg acggcgtgct gcgcaaccct ccgacgcttc
1201 gccggccgcg caggtggatc tacgcacgct cggctacagt cagcagcagc aagagaagat
1261 caaaccgaag gtgcgttcga cagtggcgca gcaccacgag gcactggtgg gccatgggtt
1321 tacacacgcg cacatcgttg cgctcagcca acacccggca gcgttaggga ccgtcgctgt
1381 cacgtatcag cacataatca cggcgttgcc agaggcgaca cacgaagaca tcgttggcgt
1441 cggcaaacag tggtccggcg cacgcgccct ggaggccttg ctcacggatg cgggggagtt
1501 gagaggtccg ccgttacagt tggacacagg ccaacttgtg aagattgcaa aacgtggcgg
1561 cgtgaccgca atggaggcag tgcatgcatc gcgcaatgca ctgacgggtg cccccctgga
1621 gacgggcgcc gctacagggc gcgtcccatt cgccattcag gctgcgcaac tgttgggaag
1681 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
1741 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
1801 gtgagcgcgc gtaatacgac tcactatagg gcgaattggg taccgggccc cccctcgagg
1861 tcctccagct tttgttccct ttagtgaggg ttaattgcgc gcttggcgta atcatggtca
1921 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
1981 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
2041 cgctcactgc ccgctttcca ccggtcgtct ccaacgacca cctcgtcgcc ttggcctgcc
2101 tcggcggacg tcctgccatg gatgcagtga aaaagggatt gccgcacgcg ccggaattga
2161 tcagaagagt caatcgccgt attggcgaac gcacgtccca tcgcgttgcc gactacgcgc
2221 aagtggttcg cgtgctggag tttttccagt gccactccca cccagcgtac gcatttgatg
2281 aggccatgac gcagttcggg atgagcagga acgggttggt acagctcttt cgcagagtgg
2341 gcgtcaccga actcgaagcc cgcggtggaa cgctcccccc agcctcgcag cgttgggacc
2401 gtatcctcca ggcatcaggg atgaaaaggg ccaaaccgtc ccctacttca gctcaaacac
2461 cggatcaggc gtctttgcat gcattcgccg attcgctgga gcgtgacctt gatgcgccca
2521 gcccaatgca cgagggagat cagacgcggg caagcagccg taaacggtcc cgatcggatc
2581 gtgctgtcac cggcccctcc gcacagcagg ctgtcgaggt gcgcgttccc gaacagcgcg
2641 atgcgctgca tttgcccctc agctggaggg taaaacgccc gcgtaccagg atctggggcg
2701 gcctcccgga tccgatatct agatcccagc tagtgaaatc tgaattggaa gagaagaaat
2761 ctgaacttag acataaattg aaatatgtgc cacatgaata tattgaattg attgaaatcg
2821 caagaaattc aactcaggat agaatccttg aaatgaaggt gatggagttc tttatgaagg
2881 tttatggtta tcgtggtaaa catttgggtg gatcaaggaa accagacgga gcaatttata
2941 ctgtcggatc tcctattgat tacggtgtga tcgttgatac taaggcatat tcaggaggtt
3001 ataatcttcc aattggtcaa gcagatgaaa tgcaaagata tgtcgaagag aatcaaacaa
3061 gaaacaagca tatcaaccct aatgaatggt ggaaagtcta tccatcttca gtaacagaat
3121 ttaagttctt gtttgtgagt ggtcatttca aaggaaacta caaagctcag cttacaagat
3181 tgaatcatat cactaattgt aatggagctg ttcttagtgt agaagagctt ttgattggtg
3241 gagaaatgat taaagctggt acattgacac ttgaggaagt gagaaggaaa tttaataacg
3301 gtgagataaa cttttaatag gagctcgaat ttccccgatc gttcaaacat ttggcaataa
3361 agtttcttaa gattgaatcc tgttgccggt cttgcgatga ttatcatata atttctgttg
3421 aattacgtta agcatgtaat aattaacatg taatgcatga cgttatttat gagatgggtt
3481 tttatgatta gagtcccgca attatacatt taatacgcga tagaaaacaa aatatagcgc
3541 gcaaactagg ataaattatc gcgcgcggtg tcatctatgt tactagatcg tgggcccaca
3601 ttcctgtaga actcggtgag acttttgttc cctttagtga gggttaattg tccaaaaaaa
3661 aaggctccaa aaggagcctt taattgtttc ggtttatgct tctccttacg catctgtgcg
3721 gtatttcaca ccgcatatcg acggtcgagg agaacttcta gtatatccac atacctaata
3781 ttattgcctt attaaaaatg gaatcccaac aattacatca aaatccacat tctcttcaaa
3841 atcaattgtc ctgtacttcc ttgttcatgt gtgttcaaaa acgttatatt tataggataa
3901 ttatactcta tttctcaaca agtaattggt tgtttggccg agcggtctaa ggcgcctgat
3961 tcaagaaata tcttgaccgc agttaactgt gggaatactc aggtatcgta agatgcaaga
4021 gttcgaatct cttagcaacc attatttttt tcctcaacat aacgagaaca cacaggggcg
4081 ctatcgcaca gaatcaaatt cgatgactgg aaattttttg ttaatttcag aggtcgcctg
4141 acgcatatac ctttttcaac tgaaaaattg ggagaaaaag gaaaggtgag aggccggaac
4201 cggcttttca tatagaatag agaagcgttc atgactaaat gcttgcatca caatacttga
4261 agttgacaat attatttaag gacctattgt tttttccaat aggtggttag caatcgtctt
4321 actttctaac ttttcttacc ttttacattt cagcaatata tatatatatt tcaaggatat
4381 accattctaa tgtctgcccc tatgtctgcc cctaagaaga tcgtcgtttt gccaggtgac
4441 cacgttggtc aagaaatcac agccgaagcc attaaggttc ttaaagctat ttctgatgtt
4501 cgttccaatg tcaagttcga tttcgaaaat catttaattg gtggtgctgc tatcgatgct
4561 acaggtgtcc cacttccaga tgaggcgctg gaagcctcca agaaggttga tgccgttttg
4621 ttaggtgctg tgggtggtcc taaatggggt accggtagtg ttagacctga acaaggttta
4681 ctaaaaatcc gtaaagaact tcaattgtac gccaacttaa gaccatgtaa ctttgcatcc
4741 gactctcttt tagacttatc tccaatcaag ccacaatttg ctaaaggtac tgacttcgtt
4801 gttgtcagag aattagtggg aggtatttac tttggtaaga gaaaggaaga cgatggtgat
4861 ggtgtcgctt gggatagtga acaatacacc gttccagaag tgcaaagaat cacaagaatg
4921 gccgctttca tggccctaca acatgagcca ccattgccta tttggtcctt ggataaagct
4981 aatgttttgg cctcttcaag attatggaga aaaactgtgg aggaaaccat caagaacgaa
5041 ttccctacat tgaaggttca acatcaattg attgattctg ccgccatgat cctagttaag
5101 aacccaaccc acctaaatgg tattataatc accagcaaca tgtttggtga tatcatctcc
5161 gatgaagcct ccgttatccc aggttccttg ggtttgttgc catctgcgtc cttggcctct
5221 ttgccagaca agaacaccgc atttggtttg tacgaaccat gccacggttc tgctccagat
5281 ttgccaaaga ataaggttga ccctatcgcc actatcttgt ctgctgcaat gatgttgaaa
5341 ttgtcattga acttgcctga agaaggtaag gccattgaag atgcagttaa aaaggttttg
5401 gatgcaggta tcagaactgg tgatttaggt ggttccaaca gtaccaccga agtcggtgat
5461 gctgtcgccg aagaagttaa gaaaatcctt gcttaaaaag attctctttt tttatgatat
5521 ttgtacataa actttataaa tgaaattcat aatagaaacg acacgaaatt acaaaatgga
5581 atatgttcat agggtagacg aaactatata cgcaatctac atacatttat caagaaggag
5641 aaaaaggagg atagtaaagg aatacaggta agcaaattga tactaatggc tcaacgtgat
5701 aaggaaaaag aattgcactt taacattaat attgacaagg aggagggcac cacacaaaaa
5761 gttaggtgta acagaaaatc atgaaactac gattcctaat ttgatattgg aggattttct
5821 ctaaaaaaaa aaaaatacaa caaataaaaa acactcaatg acctgaccat ttgatgggct
5881 caccaagctc ttaaaacggg aatttatgga tcttaattaa gacggatcgc ttgcctgtaa
5941 cttacacgcg cctcgtatct tttaatgatg gaataatttg ggaatttact ctgtgtttat
6001 ttatttttat gttttgtatt tggattttag aaagtaaata aagaaggtag aagagttacg
6061 gaatgaagaa aaaaaaataa acaaaggttt aaaaaatttc aacaaaaagc gtactttaca
6121 tatatattta ttagacaaga aaagcagatt aaatagatat acattcgatt aacgataagt
6181 aaaatgtaaa atcacaggat tttcgtgtgt ggtcttctac acagacaaga tgaaacaatt
6241 cggcattaat acctgagagc aggaagagca agataaaagg tagtatttgt tggcgatccc
6301 cctagagtct tttacatctt cggaaaacaa aaactatttt ttctttaatt tcttttttta
6361 ctttctattt ttaatttata tatttatatt aaaaaattta aattataatt atttttatag
6421 cacgtgatga aaagccttag ggtcaggtgg cacttttcgg ggaaatgtgc gatgaacccc
6481 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctaatgagac aataaccctg
6541 ataaatgctt caattatatt gaaaaaggaa gagtatgagt atccaacatt tccgtgtcgc
6601 ccttattccc ttttttgcgg ccttctgcct tcctgttttt gctcacccag aaacgctggt
6661 gaaagtaaaa gatgctgaag atcagttggg tgcccgagtg ggttacattg aactggatct
6721 caacagcggc aaaatcttag agagttttcg ccccgaagaa cgcttcccaa tgatgagcac
6781 tttcaaagtt ctgctatgtg gcgcggtgtt atcccgtatt gatgctgggc aagagcaact
6841 cggtcgccgc atacactatt ctcagaatga cttggttgag tattcaccag tcacagaaaa
6901 gcatcttacg gatggcatga cagtaagaga attatgtagt gctgccataa ccatgagtga
6961 taacactgcg gccaacttac ttctgacaac gattggagga ccgaaggagc taaccgcttt
7021 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
7081 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caactctccg
7141 caaactatta actggcgaac tacttactct tgcttcacgc cagcaactca ttgactggat
7201 ggaggcggac aaagttgcag gaccacttct gcgctcggca cttccggctg gctggtttat
7261 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
7321 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
7381 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
7441 agacgaaggg aaataaatag tagcccgcct gatgtgcggg cttttttttg gtggtggtta
7501 ccagtgtcgc ttagctaccg taggtcctct taggcggtcc gattagcgac atagtcatca
7561 atgccttgac ggcaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
7621 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
7681 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc tactaactct
7741 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
7801 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
7861 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
7921 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgctcaca
7981 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
8041 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
8101 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
8161 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
8221 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
8281 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
8341 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
8401 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcattg
8461 atgcagg
//
