| 质粒名称: | pBud-EGFP |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 23027 |
| 存储实验室: | Andrea LeBlanc |
|
载体骨架基本信息 |
|
| 载体名称: | pBudCE4.1 |
| 载体出品公司: | Invitrogen |
| 载体抗性: | 博莱霉素(Zeocin) |
| 空载体大小: | 4578 bp |
| 完整质粒大小: | 5296 bp |
| 载体类型: | 哺乳细胞表达，细胞表达 |
| 筛选标记: | 博莱霉素 |
| 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: | EGFP |
| 插入大小: | 718 bp |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | KpnI(未破坏) |
| 3' 克隆位点: | BglII(未破坏) |
| 5' 测序引物 | 未知 |
|
文献引用方法 |
|
| 材料和方法部分: | pBud-EGFP was a gift from Andrea LeBlanc (爱迪基因 plasmid # 23027) |
| 参考文献部分: | Loss of anti-Bax function in gerstmann-Straussler-Scheinker syndrome-associated prion protein mutants. Jodoin J, Misiewicz M, Makhijani P, Giannopoulos PN, Hammond J, Goodyer CG, LeBlanc AC. PLoS One. 2009 . 4(8):e6647. 10.1371/journal.pone.0006647 PubMed 19680558 |
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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 pBud-EGFP 5296 bp ds-DNA circular SYN 10-JAN-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pBud-EGFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5296)
AUTHORS Jodoin J, Misiewicz M, Makhijani P, Giannopoulos PN, Hammond J,
Goodyer CG, LeBlanc AC
TITLE Loss of anti-Bax function in gerstmann-Straussler-Scheinker
syndrome-associated prion protein mutants.
JOURNAL PLoS One. 2009 . 4(8):e6647.
PUBMED 19680558
COMMENT This file is created by Vector NTI
http://www.biofeng.com/
COMMENT ORIGDB|GenBank
COMMENT VNTAUTHORNAME|biofeng.com|
COMMENT VNTNAME|pBud-EGFP|
FEATURES Location/Qualifiers
source 1..5296
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 10..389
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 390..593
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 544..564
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 638..657
/label=T7
/note="T7 promoter, forward primer"
promoter 638..656
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 719..748
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS 764..781
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
polyA_signal 812..933
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(823..842)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 877..896
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
CDS complement(1063..1437)
/codon_start=1
/gene="Sh ble from Streptoalloteichus hindustanus"
/product="antibiotic-binding protein"
/label=BleoR
/note="confers resistance to bleomycin, phleomycin, and
Zeocin(TM)"
/translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
EFALRDPAGNCVHFVAEEQD"
promoter complement(1456..1503)
/label=EM7 promoter
/note="synthetic bacterial promoter "
promoter complement(1551..1867)
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 1565..1700
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind complement(1619..1638)
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
promoter 1889..3051
/label=EF-1-alpha promoter
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
intron 2120..3042
/label=EF-1-alpha intron A
/note="intron upstream of the start codon of human
EF-1-alpha"
primer_bind 2999..3019
/label=EF1a-F
/note="Human elongation factor-1a promoter, forward primer"
CDS 3082..3801
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(3127..3148)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(3388..3407)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 3735..3756
/label=EGFP-C
/note="EGFP, forward primer"
CDS 3828..3869
/codon_start=1
/product="epitope tag from simian virus 5"
/label=V5 tag
/translation="GKPIPNPLLGLDST"
CDS 3879..3896
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(3919..3936)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 3925..4149
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin complement(4281..4869)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4361..4380)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 4960..4978
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
primer_bind complement(5016..5038)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 5138..5157
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
ORIGIN
1 gcgcgcgttg acattgatta ttgactagtt attaatagta atcaattacg gggtcattag
61 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
121 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
181 caatagggac tttccattga cgtcaatggg tggactattt acggtaaact gcccacttgg
241 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
301 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
361 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
421 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
481 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
541 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctctctggc
601 taactagaga acccactgct tactggctta tcgaaattaa tacgactcac tatagggaga
661 cccaagcttg cattcctgca ggtcgacatc gatcttaagc agtacttcta gaggatccga
721 acaaaaactc atctcagaag aggatctgaa tatgcatacc ggtcatcatc accatcacca
781 ttgagtttga tccccgggaa ttcagacatg ataagataca ttgatgagtt tggacaaacc
841 acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta
901 tttgtaacca ttataagctg caataaacaa gttggggtgg gcgaagaact ccagcatgag
961 atccccgcgc tggaggatca tccagccggc gtcccggaaa acgattccga agcccaacct
1021 ttcatagaag gcggcggtgg aatcgaaatc tcgtagcacg tgtcagtcct gctcctcggc
1081 cacgaagtgc acgcagttgc cggccgggtc gcgcagggcg aactcccgcc cccacggctg
1141 ctcgccgatc tcggtcatgg ccggcccgga ggcgtcccgg aagttcgtgg acacgacctc
1201 cgaccactcg gcgtacagct cgtccaggcc gcgcacccac acccaggcca gggtgttgtc
1261 cggcaccacc tggtcctgga ccgcgctgat gaacagggtc acgtcgtccc ggaccacacc
1321 ggcgaagtcg tcctccacga agtcccggga gaacccgagc cggtcggtcc agaactcgac
1381 cgctccggcg acgtcgcgcg cggtgagcac cggaacggca ctggtcaact tggccatggt
1441 ttagttcctc accttgtcgt attatactat gccgatatac tatgccgatg attaattgtc
1501 aacacgtgct gatcagatcc gaaaatggat atacaagctc ccgggagctt tttgcaaaag
1561 cctaggcctc caaaaaagcc tcctcactac ttctggaata gctcagaggc agaggcggcc
1621 tcggcctctg cataaataaa aaaaattagt cagccatggg gcggagaatg ggcggaactg
1681 ggcggagtta ggggcgggat gggcggagtt aggggcggga ctatggttgc tgactaattg
1741 agatgcatgc tttgcatact tctgcctgct ggggagcctg gggactttcc acacctggtt
1801 gctgactaat tgagatgcat gctttgcata cttctgcctg ctggggagcc tggggacttt
1861 ccacaccctc gtcgagctag cttcgtgagg ctccggtgcc cgtcagtggg cagagcgcac
1921 atcgcccaca gtccccgaga agttgggggg aggggtcggc aattgaaccg gtgcctagag
1981 aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc tttttcccga
2041 gggtggggga gaaccgtata taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg
2101 gtttgccgcc agaacacagg taagtgccgt gtgtggttcc cgcgggcctg gcctctttac
2161 gggttatggc ccttgcgtgc cttgaattac ttccacctgg ctccagtacg tgattcttga
2221 tcccgagctg gagccagggg cgggccttgc gctttaggag ccccttcgcc tcgtgcttga
2281 gttgaggcct ggcctgggcg ctggggccgc cgcgtgcgaa tctggtggca ccttcgcgcc
2341 tgtctcgctg ctttcgataa gtctctagcc atttaaaatt tttgatgacc tgctgcgacg
2401 ctttttttct ggcaagatag tcttgtaaat gcgggccagg atctgcacac tggtatttcg
2461 gtttttgggc ccgcggccgg cgacggggcc cgtgcgtccc agcgcacatg ttcggcgagg
2521 cggggcctgc gagcgcggcc accgagaatc ggacgggggt agtctcaagc tggccggcct
2581 gctctggtgc ctggcctcgc gccgccgtgt atcgccccgc cctgggcggc aaggctggcc
2641 cggtcggcac cagttgcgtg agcggaaaga tggccgcttc ccggccctgc tccagggggc
2701 tcaaaatgga ggacgcggcg ctcgggagag cgggcgggtg agtcacccac acaaaggaaa
2761 agggcctttc cgtcctcagc cgtcgcttca tgtgactcca cggagtaccg ggcgccgtcc
2821 aggcacctcg attagttctg gagcttttgg agtacgtcgt ctttaggttg gggggagggg
2881 ttttatgcga tggagtttcc ccacactgag tgggtggaga ctgaagttag gccagcttgg
2941 cacttgatgt aattctcgtt ggaatttgcc ctttttgagt ttggatcttg gttcattctc
3001 aagcctcaga cagtggttca aagttttttt cttccatttc aggtgtcgtg aacacgtggt
3061 cgcggccgct tcgaaggtac catggtgagc aagggcgagg agctgttcac cggggtggtg
3121 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
3181 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
3241 ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc
3301 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
3361 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
3421 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
3481 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
3541 atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag
3601 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
3661 gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac
3721 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
3781 atggacgagc tgtacaagta gatctggccg gctgggcccg tttcgaaggt aagcctatcc
3841 ctaaccctct cctcggtctc gattctacgc gtaccggtca tcatcaccat caccattgag
3901 tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc
3961 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
4021 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
4081 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg
4141 gctctatggc ttctgaggcg gaaagaacca gtggcggtaa tacggttatc cacagaatca
4201 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
4261 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
4321 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
4381 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
4441 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
4501 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
4561 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
4621 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
4681 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc
4741 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
4801 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
4861 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
4921 tcacgttaag ggattttggt catgacatta acctataaaa ataggcgtat cacgaggccc
4981 tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
5041 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
5101 gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg
5161 agagtgcacc atatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
5221 tcaggcgcca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
5281 cttcgctatt acgcca
//
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