| 质粒名称: | pBad-EBFP2 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 14891 |
| 存储实验室: | Robert Campbell |
|
载体骨架基本信息 |
|
| 载体名称: | pBad/His B |
| 载体出品公司: | Invitrogen |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 4100 bp |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 细菌表达 |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | 低拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | EBFP2,EBFP2 blue fluorescent protein |
| 物种来源: | evolved fluorescent protein |
| 插入大小: | 720 bp |
| 基因突变: | 荧光蛋白被插入pBad/hisb的XhoI和EcoR1之间，XhoI后面有一个额外的“C”，以避免移码。 |
| GenBank编号 | EF517318 |
| 启动子 | -- |
| 标签/融合蛋白: | His（主干N端 |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | XhoI(未破坏) |
| 3' 克隆位点: | EcoRI (未破坏) |
| 5' 测序引物 | ATGCCATAGCATTTTTATCC |
| 3' 测序引物 | GATTTAATCTGTATCAGG |
|
文献引用方法 |
|
| 材料和方法部分: | pBad-EBFP2 was a gift from Robert Campbell (Addgene plasmid # 14891 ; http://n2t.net/addgene:14891 ; RRID:Addgene_14891) |
| 参考文献部分: | Exploration of New Chromophore Structures Leads to the Identification of Improved Blue Fluorescent Proteins. Ai HW, Shaner NC, Cheng Z, Tsien RY, Campbell RE. Biochemistry. 2007 Apr 20. ():. 10.1021/bi700199g PubMed 17444659 |
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 pBad-EBFP2 5429 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Bacterial expression of EBFP2 blue fluorescent protein.
ACCESSION .
VERSION .
KEYWORDS pBad-EBFP2
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5429)
AUTHORS Ai HW, Shaner NC, Cheng Z, Tsien RY, Campbell RE
TITLE Exploration of New Chromophore Structures Leads to the
Identification of Improved Blue Fluorescent Proteins.
JOURNAL Biochemistry. 2007 Apr 20. ():.
PUBMED 17444659
REFERENCE 2 (bases 1 to 5429)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5429
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS complement(53..931)
/codon_start=1
/gene="araC"
/product="L-arabinose regulatory protein"
/label=araC
/translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPPRLSHLFRQQLGIS
VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
EKVNDVAVKLS"
promoter 958..1242
/gene="araBAD"
/label=araBAD promoter
/note="promoter of the L-arabinose operon of E. coli; the
araC regulatory gene is transcribed in the opposite
direction (Guzman et al., 1995)"
primer_bind 1165..1184
/label=pBAD Forward
/note="For vectors with E. coli araBAD promoter, forward
primer"
RBS 1269..1291
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 1311..1328
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS 1332..1364
/codon_start=1
/product="leader peptide from bacteriophage T7 gene 10"
/label=T7 tag (gene 10 leader)
/note="promotes efficient translation in E. coli"
/translation="MASMTGGQQMG"
CDS 1377..1397
/codon_start=1
/product="tobacco etch virus (TEV) protease recognition and
cleavage site"
/label=TEV site
/translation="ENLYFQG"
CDS 1404..2123
/codon_start=1
/product="enhanced blue variant of GFP (Ai et al., 2007)"
/label=EBFP2
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
KFICTTGKLPVPWPTLVTTLSHGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GTYKTRAEVKFEGDTLVNRIELKGVDFKEDGNILGHKLEYNFNSHNIYIMAVKQKNGIK
VNFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDSHYLSTQSVLSKDPNEKRDHMVLL
EFRTAAGITLGMDELYK"
primer_bind complement(1449..1470)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(2169..2186)
/label=pBAD Reverse
/note="For vectors with E. coli araBAD promoter, reverse
primer"
terminator 2339..2425
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 2517..2544
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
promoter 2563..2654
/gene="bla"
/label=AmpR promoter
CDS 2655..3515
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGEPLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(2873..2892)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 3557..4012
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3644..3663)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 3854..3875
/label=F1ori-F
/note="F1 origin, forward primer"
rep_origin 4123..4711
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 4612..4631
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 4865..4882
/label=L4440
/note="L4440 vector, forward primer"
misc_feature 4897..5037
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(4963..4982)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 5123..5145
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 5283..5302
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
ORIGIN
1 tgcaaaccct atgctactcc gtcaagccgt caattgtctg attcgttacc aattatgaca
61 acttgacggc tacatcattc actttttctt cacaaccggc acggaactcg ctcgggctgg
121 ccccggtgca ttttttaaat acccgcgaga aatagagttg atcgtcaaaa ccaacattgc
181 gaccgacggt ggcgataggc atccgggtgg tgctcaaaag cagcttcgcc tggctgatac
241 gttggtcctc gcgccagctt aagacgctaa tccctaactg ctggcggaaa agatgtgaca
301 gacgcggcgg cgacaagcaa acatgctgtg cgacgctggc gatatcaaaa ttgctgtctg
361 ccaggtgatc gctgatgtac tgacaagcct cgcgtacccg attatccatc ggtggatgga
421 gcgactcgtt aatcgcttcc atgcgccgca gtaacaattg ctcaagcaga tttatcgcca
481 gcagctccga atagcgccct tccccttgcc cggcgttaat gatttgccca aacaggtcgc
541 tgaaatgcgg ctggtgcgct tcatccgggc gaaagaaccc cgtattggca aatattgacg
601 gccagttaag ccattcatgc cagtaggcgc gcggacgaaa gtaaacccac tggtgatacc
661 attcgcgagc ctccggatga cgaccgtagt gatgaatctc tcctggcggg aacagcaaaa
721 tatcacccgg tcggcaaaca aattctcgtc cctgattttt caccaccccc tgaccgcgaa
781 tggtgagatt gagaatataa cctttcattc ccagcggtcg gtcgataaaa aaatcgagat
841 aaccgttggc ctcaatcggc gttaaacccg ccaccagatg ggcattaaac gagtatcccg
901 gcagcagggg atcattttgc gcttcagcca tacttttcat actcccgcca ttcagagaag
961 aaaccaattg tccatattgc atcagacatt gccgtcactg cgtcttttac tggctcttct
1021 cgctaaccaa accggtaacc ccgcttatta aaagcattct gtaacaaagc gggaccaaag
1081 ccatgacaaa aacgcgtaac aaaagtgtct ataatcacgg cagaaaagtc cacattgatt
1141 atttgcacgg cgtcacactt tgctatgcca tagcattttt atccataaga ttagcggata
1201 ctacctgacg ctttttatcg caactctcta ctgtttctcc atacccgttt ttttgggcta
1261 gaaataattt tgtttaactt taagaaggag atatacatat gcggggttct catcatcatc
1321 atcatcatgg tatggctagc atgactggtg gacagcaaat gggtcgggat ctgtacgaga
1381 acctgtactt ccagggctcg agcatggtga gcaagggcga ggagctgttc accggggtgg
1441 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgaggggcg
1501 agggcgaggg cgatgccacc aacggcaagc tgaccctgaa gttcatctgc accaccggca
1561 agctgcccgt gccctggccc accctcgtga ccaccctgag ccacggcgtg cagtgcttcg
1621 cccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
1681 acgtccagga gcgcaccatc ttcttcaagg acgacggcac ctacaagacc cgcgccgagg
1741 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcgtc gacttcaagg
1801 aggacggcaa catcctgggg cacaagctgg agtacaactt caacagccac aacatctata
1861 tcatggccgt caagcagaag aacggcatca aggtgaactt caagatccgc cacaacgtgg
1921 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
1981 ccgtgctgct gcccgacagc cactacctga gcacccagtc cgtgctgagc aaagacccca
2041 acgagaagcg cgatcacatg gtcctgctgg agttccgcac cgccgccggg atcactctcg
2101 gcatggacga gctgtacaag taagaattcg aagcttggct gttttggcgg atgagagaag
2161 attttcagcc tgatacagat taaatcagaa cgcagaagcg gtctgataaa acagaatttg
2221 cctggcggca gtagcgcggt ggtcccacct gaccccatgc cgaactcaga agtgaaacgc
2281 cgtagcgccg atggtagtgt ggggtctccc catgcgagag tagggaactg ccaggcatca
2341 aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt
2401 gaacgctctc ctgagtagga caaatccgcc gggagcggat ttgaacgttg cgaagcaacg
2461 gcccggaggg tggcgggcag gacgcccgcc ataaactgcc aggcatcaaa ttaagcagaa
2521 ggccatcctg acggatggcc tttttgcgtt tctacaaact cttttgttta tttttctaaa
2581 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
2641 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
2701 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
2761 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
2821 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
2881 gcgcggtatt atcccgtgtt gacgccgggc aagagcaact cggtcgccgc atacactatt
2941 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
3001 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
3061 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
3121 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
3181 gtgacaccac gatgcctgca gcaatggcaa caacgttgcg caaactatta actggcgaac
3241 cacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
3301 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
3361 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
3421 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
3481 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
3541 tactttagat tgatttacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
3601 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
3661 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
3721 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga tttgggtgat
3781 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
3841 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcgggc
3901 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
3961 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat ttaaaaggat
4021 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
4081 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
4141 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
4201 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
4261 aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
4321 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
4381 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
4441 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
4501 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
4561 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
4621 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
4681 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
4741 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
4801 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
4861 gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg atgcggtatt ttctccttac
4921 gcatctgtgc ggtatttcac accgcatctg gtgcactctc agtacaatct gctctgatgc
4981 cgcatagtta agccagtata cactccgcta tcgctacgtg actgggtcat ggctgcgccc
5041 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
5101 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
5161 ccgaaacgcg cgaggcagaa ggagatggcg cccaacagtc ccccggccac ggggcctgcc
5221 accataccca cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg atcttcccca
5281 tcggtgatgt cggcgatata ggcgccagca accgcacctg tggcgccggt gatgccggcc
5341 acgatgcgtc cggcgtagag gatctgctca tgtttgacag cttatcatcg atgcataatg
5401 tgcctgtcaa atggacgaag cagggattc
//
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