| 载体基本信息 | |||
| 载体名称 | pEU-GST-FV | 载体抗性 | Ampicillin |
|---|---|---|---|
| 载体长度 | 5612 bp | 载体类型 | Structural Genomics Vectors |
| 载体来源 | Vinarov DA, Lytle BL, Peterson FC, Tyler EM, Volkman BF, Markley JL. | 拷贝数 | High copy number |
| 3'引物 | M13 rev | ||
pEU-GST-FV 载体载体图谱
pEU-GST-FV 载体载体序列
LOCUS 40924_18636 5612 bp DNA circular SYN 01-JAN-1980
DEFINITION Flexi(R) Vector with the barnase gene, for in vitro transcription
and translation of a protein with an N-terminal GST-HRV 3C cassette.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5612)
AUTHORS Vinarov DA, Lytle BL, Peterson FC, Tyler EM, Volkman BF, Markley JL.
TITLE Cell-free protein production and labeling protocol for NMR-based
structural proteomics.
JOURNAL Nat. Methods 2004;1:149-53.
PUBMED 15782178
REFERENCE 2 (bases 1 to 5612)
AUTHORS Center for Eukaryotic Structural Genomics
TITLE Direct Submission
REFERENCE 3 (bases 1 to 5612)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nat.
Methods"; date: "2004"; volume: "1"; pages: "149-53"
COMMENT SGRef: number: 2; type: "Journal Article"
COMMENT To remove the lethal barnase gene, insert an ORF between the SgfI
site and the PmeI site or a downstream site.
FEATURES Location/Qualifiers
source 1..5612
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 1..19
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
CDS 155..805
/codon_start=1
/label=GST
/note="glutathione S-transferase from Schistosoma
japonicum"
/translation="SPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKKF
ELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRYG
VSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVLY
MDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
CDS 812..835
/codon_start=1
/label=HRV 3C site
/note="recognition and cleavage site for human rhinovirus
3C and PreScission proteases"
/translation="LEVLFQGP"
CDS 867..1199
/codon_start=1
/label=barnase
/note="ribonuclease from Bacillus amyloliquefaciens"
/translation="MAQVINTFDGVADYLQTYHKLPDNYITKSEAQALGWVASKGNLAD
VAPGKSIGGDIFSNREGKLPGKSGRTWREADINYTSGFRNSDRILYSSDWLIYKTTDHY
QTFTKIR"
promoter 1218..1248
/label=lac UV5 promoter
/note="E. coli lac promoter with an 'up' mutation"
CDS 1302..1958
/codon_start=1
/label=CmR
/note="chloramphenicol acetyltransferase"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
misc_feature 1971..2026
/label=MCS
/note="pUC18/19 multiple cloning site"
primer_bind complement(2697..2713)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(2721..2737)
/label=lac operator
/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(2745..2775)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(2790..2811)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(3099..3687)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(3861..4718)
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(4719..4823)
/label=AmpR promoter
我们不提供该载体，载体信息仅供查询和参考。