| 质粒名称: | pAC1402-pCR8-Clover_NLSPUFa |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 71900 |
| 存储实验室: | Albert Cheng |
|
载体骨架基本信息 |
|
| 载体名称: | pCR8/GW/TOPO |
| 载体出品公司: | Invitrogen |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | Gateway Donor |
| 筛选标记: | -- |
| 载体抗性: | 大观霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | -- |
|
插入基因信息 |
|
| 插入基因名称: | Clover_PUFa |
| 物种来源: | -- |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子: | -- |
| 标签/融合蛋白: | Clover（插入件上的N端子） |
| 克隆方法: | -- |
| 5’克隆位点: | -- |
| 3’克隆位点: | -- |
| 5’测序引物 | M13F |
| 3’测序引物 | M13R |
|
文献引用方法 |
|
| 材料和方法部分: | pAC1402-pCR8-Clover_NLSPUFa was a gift from Albert Cheng (Addgene plasmid # 71900 ; http://n2t.net/addgene:71900 ; RRID:Addgene_71900) |
| 参考文献部分: | Casilio: a versatile CRISPR-Cas9-Pumilio hybrid for gene regulation and genomic labeling. Cheng AW, Jillette N, Lee P, Plaskon D, Fujiwara Y, Wang W, Taghbalout A, Wang H. Cell Res. 2016 Feb;26(2):254-7. doi: 10.1038/cr.2016.3. Epub 2016 Jan 15. 10.1038/cr.2016.3 PubMed 26768771 |
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 pAC1402-pCR8-Clover_NLSPUFa 4694 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Clover_NLSPUFa in pCR8 gateway donor vector.
ACCESSION .
VERSION .
KEYWORDS pAC1402-pCR8-Clover_NLSPUFa
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4694)
AUTHORS Cheng AW, Jillette N, Lee P, Plaskon D, Fujiwara Y, Wang W,
Taghbalout A, Wang H
TITLE Casilio: a versatile CRISPR-Cas9-Pumilio hybrid for gene regulation
and genomic labeling.
JOURNAL Cell Res. 2016 Feb;26(2):254-7. doi: 10.1038/cr.2016.3. Epub 2016
Jan 15.
PUBMED 26768771
REFERENCE 2 (bases 1 to 4694)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..4694
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 92..109
/label=L4440
/note="L4440 vector, forward primer"
terminator 268..295
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
terminator 387..473
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
primer_bind 522..544
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 536..553
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 537..553
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 569..668
/gene="mutant version of attL"
/label=attL1
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
regulatory 686..695
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 692..1408
/codon_start=1
/product="bright green-yellow fluorescent protein derived
from GFP (Lam et al., 2012)"
/label=Clover
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
KFICTTGKLPVPWPTLVTTFGYGVACFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDD
GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIK
ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSHQSALSKDPNEKRDHMVLL
EFVTAAGITHGMDELYK"
primer_bind complement(737..758)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind 1345..1366
/label=EGFP-C
/note="EGFP, forward primer"
promoter complement(1424..1442)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(1425..1442)
/label=SP6
/note="SP6 promoter, forward primer"
CDS 1481..1501
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
protein_bind complement(2573..2672)
/gene="mutant version of attL"
/label=attL2
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
primer_bind complement(2689..2708)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(2690..2708)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(2713..2729)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(2713..2729)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
CDS 3160..3951
/codon_start=1
/gene="aadA"
/product="aminoglycoside adenylyltransferase (Murphy,
1985)"
/label=SmR
/note="confers resistance to spectinomycin and
streptomycin"
/translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
rep_origin 4044..4632
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 4533..4552
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
661 agcaggctcc gaattcaccg gtgccgccac catggtgagc aagggcgagg agctgttcac
721 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
781 ccgcggcgag ggcgagggcg atgccaccaa cggcaagctg accctgaagt tcatctgcac
841 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accttcggct acggcgtggc
901 ctgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
961 cgaaggctac gtccaggagc gcaccatctc tttcaaggac gacggtacct acaagacccg
1021 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
1081 cttcaaggag gacggcaaca tcctggggca caagctggag tacaacttca acagccacaa
1141 cgtctatatc acggccgaca agcagaagaa cggcatcaag gctaacttca agatccgcca
1201 caacgttgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg
1261 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc catcagtccg ccctgagcaa
1321 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat
1381 tacacatggc atggacgagc tgtacaagtc tagagggccc tattctatag tgtcacctaa
1441 atgcggaggc ggcggtagcg ggccggccgg gatcctcccc cccaagaaaa agaggaaggt
1501 atctagaggc cgcagccgcc ttttggaaga ttttcgaaac aaccggtacc ccaatttaca
1561 actgcgggag attgctggac atataatgga attttcccaa gaccagcatg ggtccagatt
1621 cattcagctg aaactggagc gtgccacacc agctgagcgc cagcttgtct tcaatgaaat
1681 cctccaggct gcctaccaac tcatggtgga tgtgtttggt aattacgtca ttcagaagtt
1741 ctttgaattt ggcagtcttg aacagaagct ggctttggca gaacggattc gaggccacgt
1801 cctgtcattg gcactacaga tgtatggcag ccgtgttatc gagaaagctc ttgagtttat
1861 tccttcagac cagcagaatg agatggttcg ggaactagat ggccatgtct tgaagtgtgt
1921 gaaagatcag aatggcaatc acgtggttca gaaatgcatt gaatgtgtac agccccagtc
1981 tttgcaattt atcatcgatg cgtttaaggg acaggtattt gccttatcca cacatcctta
2041 tggctgccga gtgattcaga gaatcctgga gcactgtctc cctgaccaga cactccctat
2101 tttagaggag cttcaccagc acacagagca gctggtacag gatcaatatg gaaattatgt
2161 aatccaacat gtactggagc acggtcgtcc tgaggataaa agcaaaattg tagcagaaat
2221 ccgaggcaat gtacttgtat tgagtcagca caaatttgca agcaatgttg tggagaagtg
2281 tgttactcac gcctcacgta cggagcgcgc tgtgctcatc gatgaggtgt gcaccatgaa
2341 cgacggtccc cacagtgcct tatacaccat gatgaaggac cagtatgcca actacgtggt
2401 ccagaagatg attgacgtgg cggagccagg ccagcggaag atcgtcatgc ataagatccg
2461 gccccacatc gcaactcttc gtaagtacac ctatggcaag cacattctgg ccaagctgga
2521 gaagtactac atgaagaacg gtgttgactt agggtgatta attaagaatt cgacccagct
2581 ttcttgtaca aagttggcat tataaaaaat aattgctcat caatttgttg caacgaacag
2641 gtcactatca gtcaaaataa aatcattatt tgccatccag ctgatatccc ctatagtgag
2701 tcgtattaca tggtcatagc tgtttcctgg cagctctggc ccgtgtctca aaatctctga
2761 tgttacattg cacaagataa aaatatatca tcatgcctcc tctagaccag ccaggacaga
2821 aatgcctcga cttcgctgct gcccaaggtt gccgggtgac gcacaccgtg gaaacggatg
2881 aaggcacgaa cccagtggac ataagcctgt tcggttcgta agctgtaatg caagtagcgt
2941 atgcgctcac gcaactggtc cagaaccttg accgaacgca gcggtggtaa cggcgcagtg
3001 gcggttttca tggcttgtta tgactgtttt tttggggtac agtctatgcc tcgggcatcc
3061 aagcagcaag cgcgttacgc cgtgggtcga tgtttgatgt tatggagcag caacgatgtt
3121 acgcagcagg gcagtcgccc taaaacaaag ttaaacatca tgagggaagc ggtgatcgcc
3181 gaagtatcga ctcaactatc agaggtagtt ggcgtcatcg agcgccatct cgaaccgacg
3241 ttgctggccg tacatttgta cggctccgca gtggatggcg gcctgaagcc acacagtgat
3301 attgatttgc tggttacggt gaccgtaagg cttgatgaaa caacgcggcg agctttgatc
3361 aacgaccttt tggaaacttc ggcttcccct ggagagagcg agattctccg cgctgtagaa
3421 gtcaccattg ttgtgcacga cgacatcatt ccgtggcgtt atccagctaa gcgcgaactg
3481 caatttggag aatggcagcg caatgacatt cttgcaggta tcttcgagcc agccacgatc
3541 gacattgatc tggctatctt gctgacaaaa gcaagagaac atagcgttgc cttggtaggt
3601 ccagcggcgg aggaactctt tgatccggtt cctgaacagg atctatttga ggcgctaaat
3661 gaaaccttaa cgctatggaa ctcgccgccc gactgggctg gcgatgagcg aaatgtagtg
3721 cttacgttgt cccgcatttg gtacagcgca gtaaccggca aaatcgcgcc gaaggatgtc
3781 gctgccgact gggcaatgga gcgcctgccg gcccagtatc agcccgtcat acttgaagct
3841 agacaggctt atcttggaca agaagaagat cgcttggcct cgcgcgcaga tcagttggaa
3901 gaatttgtcc actacgtgaa aggcgagatc accaaggtag tcggcaaata accctcgagc
3961 cacccatgac caaaatccct taacgtgagt tacgcgtcgt tccactgagc gtcagacccc
4021 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
4081 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
4141 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg
4201 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
4261 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
4321 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
4381 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagcattga
4441 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
4501 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
4561 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
4621 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
4681 tttgctcaca tgtt
//
