| 质粒名称: | pTAL3 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 31034 |
| 存储实验室: | Adam Bogdanove, Daniel Voytas |
|
载体骨架基本信息 |
|
| 载体名称: | pDW1789 |
| 载体抗性: | 氨苄青霉素 (50 ug/ml) |
| 空载体大小: | 5582 bp |
| 筛选标记: | HIS3 |
| 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(未破坏) |
| 3' 克隆位点: | BamHI (未破坏) |
| 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: | pTAL3 was a gift from Adam Bogdanove & Daniel Voytas (Addgene plasmid # 31034 ; http://n2t.net/addgene:31034 ; RRID:Addgene_31034) |
| 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 |
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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 pTAL3 7530 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pTAL3
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7530)
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 7530)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7530
/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)"
CDS complement(3990..4649)
/codon_start=1
/gene="S. cerevisiae HIS3"
/product="imidazoleglycerol-phosphate dehydratase, required
for histidine biosynthesis"
/label=HIS3
/note="yeast auxotrophic marker"
/translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
KEALLARGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHFL
ESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
promoter complement(4650..4837)
/gene="S. cerevisiae HIS3"
/label=HIS3 promoter
misc_feature 4988..5491
/label=CEN/ARS
/note="S. cerevisiae CEN6 centromere fused to an
autonomously replicating sequence"
promoter 5546..5637
/gene="bla"
/label=AmpR promoter
rep_origin 6705..7293
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 7194..7213
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 7447..7464
/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 taggacacca ttgattaacg
3721 cttacaattt cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
3781 tagatccgtc gagttcaaga gaaaaaaaaa gaaaaagcaa aaagaaaaaa ggaaagcgcg
3841 cctcgttcag aatgacacgt atagaatgat gcattacctt gtcatcttca gtatcatact
3901 gttcgtataa atacttactg acattcatag ctatacatat atacacatgt atatatatcg
3961 tatgctggag ctttaaataa tcggtgtcac tacataagaa cacctttggt ggagggaaca
4021 tcgttggttc cattgggcga ggtggcttct cttatggcaa ccgcaagagc cttgaacgca
4081 ctctcactac ggtgatggtc attcttgcct cgcagacaat caacgtggag ggtaattctg
4141 cttgcctctg caaaactttc aagaaaatgc gggatcatct cgcaagagag gtctcctact
4201 ttctcccttt gcaaaccaag ttcgacaact gcgtatggcc tgttagaaag gtctaccacc
4261 gctctggaaa gtgcctcatc caaaggcgca aatcctgatc caaacctttt tactccacgc
4321 gccagtagag cctctttaaa agcctgaccg agagcaatcc cgcagtcttc agtggtgtga
4381 tggtcgtcta tgtgtaagtc accaatgcac tcaacgatta gcgaccagcc ggaatgcttg
4441 gccagagcat gtatcatgtg gtccagaaac cctatacctg tgtggacgtt aatcacttgc
4501 gattgtgtgg cctgttctgc tactgcttct gcctcttttt ctgggaagat cgagtgctct
4561 atcgctaggg gaccaccctt taaagagata gcaatctgaa tcttggtttc atttgtaata
4621 cgctttacta gggctttctg ctctgtcatc tttgccttcg tttatcttgc ctgctcattt
4681 tttagtatat tcttcgaaga aatcacatta ctttatataa tgtataattc attatgtgat
4741 aatgccaatc gctaagaaaa aaaaagagtc atccgctagg tggaaaaaaa aaaatgaaaa
4801 tcattaccga ggcataaaaa aatatagagt gtactagagg aggccaagag taatagaaaa
4861 agaaaattgc gggaaaggac tgtgttatga cttccctgac taatgccgtg ttcaaacgat
4921 acctggcagt gactcctagc gctcaccaag ctcttaaaac gggaatttat ggatcttaat
4981 taagacggat cgcttgcctg taacttacac gcgcctcgta tcttttaatg atggaataat
5041 ttgggaattt actctgtgtt tatttatttt tatgttttgt atttggattt tagaaagtaa
5101 ataaagaagg tagaagagtt acggaatgaa gaaaaaaaaa taaacaaagg tttaaaaaat
5161 ttcaacaaaa agcgtacttt acatatatat ttattagaca agaaaagcag attaaataga
5221 tatacattcg attaacgata agtaaaatgt aaaatcacag gattttcgtg tgtggtcttc
5281 tacacagaca agatgaaaca attcggcatt aatacctgag agcaggaaga gcaagataaa
5341 aggtagtatt tgttggcgat ccccctagag tcttttacat cttcggaaaa caaaaactat
5401 tttttcttta atttcttttt ttactttcta tttttaattt atatatttat attaaaaaat
5461 ttaaattata attattttta tagcacgtga tgaaaagcct tagggtcagg tggcactttt
5521 cggggaaatg tgcgatgaac ccctatttgt ttatttttct aaatacattc aaatatgtat
5581 ccgctaatga gacaataacc ctgataaatg cttcaattat attgaaaaag gaagagtatg
5641 agtatccaac atttccgtgt cgcccttatt cccttttttg cggccttctg ccttcctgtt
5701 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcccga
5761 gtgggttaca ttgaactgga tctcaacagc ggcaaaatct tagagagttt tcgccccgaa
5821 gaacgcttcc caatgatgag cactttcaaa gttctgctat gtggcgcggt gttatcccgt
5881 attgatgctg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
5941 gagtattcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgt
6001 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgattgga
6061 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
6121 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
6181 gtagcaatgg caacaactct ccgcaaacta ttaactggcg aactacttac tcttgcttca
6241 cgccagcaac tcattgactg gatggaggcg gacaaagttg caggaccact tctgcgctcg
6301 gcacttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
6361 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
6421 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
6481 ctgattaagc attggtaact gtcagacgaa gggaaataaa tagtagcccg cctgatgtgc
6541 gggctttttt ttggtggtgg ttaccagtgt cgcttagcta ccgtaggtcc tcttaggcgg
6601 tccgattagc gacatagtca tcaatgcctt gacggcaaaa tcccttaacg tgagttttcg
6661 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
6721 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
6781 ccggatcaag agctactaac tctttttccg aaggtaactg gcttcagcag agcgcagata
6841 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
6901 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
6961 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
7021 tgaacggggg gttcgtgctc acagcccagc ttggagcgaa cgacctacac cgaactgaga
7081 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
7141 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
7201 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
7261 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
7321 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
7381 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
7441 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
7501 cccgcgcgtt ggccgattca ttgatgcagg
//
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