| 质粒名称: | FLAG-EAF pBacPAK8 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 15323 |
| 存储实验室: | Joan Conaway, Ronald Conaway |
|
载体骨架基本信息 |
|
| 载体名称: | pBacPAK8 |
| 载体出品公司: | Clontech |
| 载体抗性: | 氨苄霉素 |
| 空载体大小: | 5538 bp |
| 完整质粒大小: | 6345 bp |
| 载体类型: | Baculovirus construction |
| 高拷贝/低拷贝: | 未知 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | ELL Associated Factor |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 807 bp |
| 基因突变: | EAF;EAF1 |
| 启动子 | polyhedrin |
| 标签/融合蛋白: |
6x His (N terminal on backbone)
FLAG (N terminal on backbone)
|
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | Xho1(未破坏) |
| 3' 克隆位点: | Kpn1(未破坏) |
| 5' 测序引物 | Bac1 |
| 3' 测序引物 | Bac2 |
|
文献引用方法 |
|
| 材料和方法部分: | FLAG-EAF pBacPAK8 was a gift from Joan Conaway & Ronald Conaway (爱迪基因 plasmid # 15323) |
| 参考文献部分: | ELL-associated factors 1 and 2 are positive regulators of RNA polymerase II elongation factor ELL. Kong SE, Banks CA, Shilatifard A, Conaway JW, Conaway RC. Proc Natl Acad Sci U S A. 2005 Jul 19. 102(29):10094-8. 10.1073/pnas.0503017102 PubMed 16006523 |
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 FLAG-EAF pBacPAK8 6252 bp ds-DNA circular SYN 10-JAN-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS FLAG-EAF pBacPAK8
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6252)
AUTHORS Kong SE, Banks CA, Shilatifard A, Conaway JW, Conaway RC
TITLE ELL-associated factors 1 and 2 are positive regulators of RNA
polymerase II elongation factor ELL.
JOURNAL Proc Natl Acad Sci U S A. 2005 Jul 19. 102(29):10094-8.
PUBMED 16006523
COMMENT This file is created by Vector NTI
http://www.biofeng.com/
COMMENT ORIGDB|GenBank
COMMENT VNTAUTHORNAME|biofeng.com|
COMMENT VNTNAME|FLAG-EAF pBacPAK8|
FEATURES Location/Qualifiers
source 1..6252
/organism="synthetic DNA construct"
/mol_type="other DNA"
misc_recomb 2..829
/label=baculovirus recombination region (lef2/ORF603)
/note="contains ORF603 and part of lef2"
CDS 2..126
/codon_start=3
/product="baculovirus late expression factor 2"
/label=lef2
/translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
CDS complement(164..769)
/codon_start=1
/product="baculovirus ORF603 protein"
/label=ORF603
/translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV
IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI
KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ
INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR"
promoter 833..924
/gene="polh from Autographa californica"
/label=polyhedrin promoter
/note="promoter for the baculovirus polyhedrin gene"
primer_bind 849..866
/label=Polyhedrin-F
/note="Polyhedrin promoter, forward primer"
CDS 934..951
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS 1006..1029
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
misc_recomb 1890..3247
/label=baculovirus recombination region (ORF1629)
/note="contains part of ORF1629"
CDS complement(1906..3247)
/codon_start=2
/product="baculovirus capsid-associated protein"
/label=ORF1629
/note="required for viral replication"
/translation="PSLRQSLYNTIAYIERLLNIGTVNDSEITMLIADFYDLYSNYNIE
LPPPQALPRSRRPSVVQPAAPAPVPTIVREQTKPEQIIPAAPPPPPSPVPNIPAPPPPP
PPSMSELPPAPPMPTEPQPAAPLDDRQQLLEAIRNEKNRTRLRPVKPKTAPETSTIVEV
PTVLPKETFEPKPPSASPPPPPPPPPPPAPPAPPPMVDLSSAPPPPPLVDLPSEMLPPP
APSLSNVLSELKSGTVRLKPAQKRPQSEIIPKSSTTNLIADVLADTINRRRVAMAKSSS
EATSNDEGWDDDDNRPNKANTPDVKYVQALFNVFTSSQLYTNDSDERNTKAHNILNDVE
PLLQNKTQTNIDKARLLLQDLASFVALSENPLDSPAIGSEKQPLFETNRNLFYKSIEDL
IFKFRYKDAENHLIFALTYHPKDYKFNELLKYVQQLSVNQQRTESSA"
rep_origin 3394..3849
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3481..3500)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 3691..3712
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(3866..3885)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 3985..4007
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(4045..4063)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 4131..4235
/gene="bla"
/label=AmpR promoter
CDS 4236..5096
/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 complement(4454..4473)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 5267..5855
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5756..5775
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 6009..6026
/label=L4440
/note="L4440 vector, forward primer"
ORIGIN
1 atgcaccttt taatcaaatc ccaagatgtg tataaaccac caaactgcca aaaaatgaaa
61 actgtcgata agctctgtcc gtttgctggc aactgcaagg gtctcaatcc tatttgtaat
121 tattgaataa taaaacaatt ataaatgtca aatttgtttt ttattaacga tacaaaccaa
181 acgcaacaag aacatttgta gtattatcta taattgaaaa agcgtagtta taatcgctga
241 ggtaatattt aaaatcattt tcaaatgatt cacagttaat ttgcgacaat ataattttat
301 tttcacataa actagacgcc ttgtcgtctt cttcttcgta ttccttctct ttttcatttt
361 tctcctcata aaaattaaca tagttattat cgtatccata tatgtatcta tcgtatagag
421 taaatttttt gttgtcataa atatatatgt cttttttaat ggggtgtata gtaccgctgc
481 gcatagtttt tctgtaattt acaacagtgc tattttctgg tagttcttcg gagtgtgttg
541 ctttaattat taaatttata taatcaatga atttgggatc gtcggttttg tacaatatgt
601 tgccggcata gtacgcagct tcttctagtt caattacacc attttttagc agcaccggat
661 taacataact ttccaaaatg ttgtacgaac cgttaaacaa aaacagttca cctccctttt
721 ctatactatt gtctgcgagc agttgtttgt tgttaaaaat aacagccatt gtaatgagac
781 gcacaaacta atatcacaaa ctggaaatgt ctatcaatat atagttgctg atatcatgga
841 gataattaaa atgataacca tctcgcaaat aaataagtat tttactgttt tcgtaacagt
901 tttgtaataa aaaaacctat aaataccatg ggtcatcatc atcaccatca cggtggtcga
961 caagcttgga tccctgcagg cctcgagttc gaatctagaa tgccagacta caaggacgat
1021 gatgacaagc tcgagatgaa tgggaccgca aacccgctgc tggaccgcga ggaacattgc
1081 ctgaggctcg gggagagctt cgagaagcgg ccgcgggcct ccttccacac tattcgttat
1141 gattttaaac cagcatctat agacacttcc tgtgaaggag agcttcaagt tggcaaagga
1201 gatgaagtca caattacact gccacatatc cctggatcca caccacccat gactgtgttc
1261 aaggggaaca aacggcctta ccagaaagac tgtgtgctta ttattaatca tgacactggt
1321 gaatatgtgc tggaaaaact cagtagcagc attcaggtga agaaaacaag agctgagggc
1381 agcagtaaaa tccaggcccg aatggaacag cagcccactc gtcctccaca gacgtcacag
1441 ccaccaccac ctccaccacc tatgccattc agagctccaa cgaagcctcc agttggaccc
1501 aaaacttctc ccttgaaaga taacccctca cctgaacctc agttggatga catcaaaaga
1561 gagctgaggg ctgaagttga cattattgaa caaatgagca gcagcagtgg gagcagctct
1621 tcagactctg agagctcttc gggaagtgat gacgatagct ccagcagtgg aggcgaggac
1681 aatggcccag cctctcctcc gcagccttca caccagcagc cctacaacag taggcctgcc
1741 gttgccaatg gaaccagccg gccacaagga agcaaccagc tcatgaacac cctcagaaat
1801 gacttgcagt tgagtgagtc tggcagtgac agtgatgact agggtaccga gctcgaattc
1861 ccgggcggcc gcttaattaa ttgatccggg ttattagtac atttattaag cgctagattc
1921 tgtgcgttgt tgatttacag acaattgttg tacgtatttt aataattcat taaatttata
1981 atctttaggg tggtatgtta gagcgaaaat caaatgattt tcagcgtctt tatatctgaa
2041 tttaaatatt aaatcctcaa tagatttgta aaataggttt cgattagttt caaacaaggg
2101 ttgtttttcc gaaccgatgg ctggactatc taatggattt tcgctcaacg ccacaaaact
2161 tgccaaatct tgtagcagca atctagcttt gtcgatattc gtttgtgttt tgttttgtaa
2221 taaaggttcg acgtcgttca aaatattatg cgcttttgta tttctttcat cactgtcgtt
2281 agtgtacaat tgactcgacg taaacacgtt aaataacgct tggacatatt taacatcggg
2341 cgtgttagct ttattaggcc gattatcgtc gtcgtcccaa ccctcgtcgt tagaagttgc
2401 ttccgaagac gattttgcca tagccacacg acgcctatta attgtgtcgg ctaacacgtc
2461 cgcgatcaaa tttgtagttg agctttttgg aattatttct gattgcgggc gtttttgggc
2521 gggtttcaat ctaactgtgc ccgattttaa ttcagacaac acgttagaaa gcgatggtgc
2581 aggcggtggt aacatttcag acggcaaatc tactaatggc ggcggtggtg gagctgatga
2641 taaatctacc atcggtggag gcgcaggcgg ggctggcggc ggaggcggag gcggaggtgg
2701 tggcggtgat gcagacggcg gtttaggctc aaatgtctct ttaggcaaca cagtcggcac
2761 ctcaactatt gtactggttt cgggcgccgt ttttggtttg accggtctga gacgagtgcg
2821 atttttttcg tttctaatag cttccaacaa ttgttgtctg tcgtctaaag gtgcagcggg
2881 ttgaggttcc gtcggcattg gtggagcggg cggcaattca gacatcgatg gtggtggtgg
2941 tggtggaggc gctggaatgt taggcacggg agaaggtggt ggcggcggtg ccgccggtat
3001 aatttgttca ggtttagttt gttcgcgcac gattgtgggc accggcgcag gcgccgctgg
3061 ctgcacaacg gaaggtcgtc tgcttcgagg cagcgcttgg ggtggtggca attcaatatt
3121 ataattggaa tacaaatcgt aaaaatctgc tataagcatt gtaatttcgc tatcgtttac
3181 cgtgccgata tttaacaacc gctcaatgta agcaattgta ttgtaaagag attgtctcaa
3241 gctcggatcg atcccgcacg ccgataacaa gccttttcat ttttactaca gcattgtagt
3301 ggcgagacac ttcgctgtcg tcgcctgatg cggtattttc tccttacgca tctgtgcggt
3361 atttcacacc gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg
3421 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
3481 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
3541 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
3601 aacttgattt gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
3661 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
3721 tcaaccctat ctcgggctat tcttttgatt tataagggat tttgccgatt tcggcctatt
3781 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt
3841 ttacaatttt atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc
3901 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
3961 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
4021 caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca
4081 tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
4141 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
4201 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
4261 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
4321 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
4381 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
4441 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
4501 tcggtcgccg catacactat tctcagaatg acttggttga atattcacca gtcacagaaa
4561 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
4621 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
4681 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
4741 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
4801 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
4861 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
4921 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
4981 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
5041 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
5101 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
5161 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
5221 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
5281 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
5341 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
5401 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
5461 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
5521 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
5581 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
5641 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
5701 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
5761 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
5821 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
5881 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
5941 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
6001 ccgagcgcag cgagtcagtg agcgaggaag ctcgacatgc tgaacaacaa gattaatatg
6061 cctccgtgta taaaaaaaat attgaacgat ttgaaagaaa acaatgtacc gcgcggcggt
6121 atgtacagga agaggtttat actaaactgt tacattgcaa acgtggtttc gtgtgccaag
6181 tgtgaaaacc gatgtttaat caaggctctg acgcatttct acaaccacga ctccaagtgt
6241 gtcggtgaag tc
//
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