| 载体基本信息 |
| 载体名称 |
pMETα A |
载体抗性 |
Ampicillin |
| 载体长度 |
8045 bp |
载体类型 |
Yeast Expression Vectors |
| 筛选标记 |
ADE2 |
拷贝数 |
High copy number |
| 启动子 |
44774 |
克隆方法 |
Enzyme digestion and ligation |
| 5引物 |
AUG1-F: 5´-CAATTTACATCTTTATTTATTAACG-3´; alpha-Factor-F: 5´-TACTATTGCCAGCATTGCTGC-3´ |
3引物 |
AUG1-R: 5´-GAAGAGAAAAACATTAGTTGGC-3´ |
| 载体标签 |
C-V5, C-His |
pMETα A 载体载体图谱
pMETα A 载体载体序列
LOCUS Exported 8045 bp ds-DNA circular SYN 13-JAN-2022
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8045)
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..8045
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS 1197..1463
/codon_start=1
/gene="MF-alpha-1"
/product="N-terminal secretion signal from S. cerevisiae
alpha-factor"
/note="alpha-factor secretion signal"
/note="Cleavage by the Kex2 protease occurs after the
dibasic KR sequence. The EA dipeptides are then removed by
dipeptidyl aminopeptidase A."
/translation="MRFPSIFTAVLFAASSALAAPVNTTTEDETAQIPAEAVIGYSDLE
GDFDVAVLPFSNSTNNGLLFINTTIASIAAKEEGVSLEKREAEA"
CDS 1534..1575
/codon_start=1
/product="epitope tag from simian virus 5"
/note="V5 tag"
/translation="GKPIPNPLLGLDST"
CDS 1585..1602
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis"
/translation="HHHHHH"
promoter 2710..3085
/gene="S. cerevisiae ADE2"
/note="ADE2 promoter"
CDS 3086..4801
/codon_start=1
/gene="S. cerevisiae ADE2"
/product="phosphoribosylaminoimidazole carboxylase,
required for adenine biosynthesis"
/note="ADE2"
/note="yeast auxotrophic marker"
/translation="MDSRTVGILGGGQLGRMIVEAANRLNIKTVILDAENSPAKQISNS
NDHVNGSFSNPLDIEKLAEKCDVLTIEIEHVDVPTLKNLQVKHPKLKIYPSPETIRLIQ
DKYIQKEHLIKNGIAVTQSVPVEQASETSLLNVGRDLGFPFVLKSRTLAYDGRGNFVVK
NKEMIPEALEVLKDRPLYAEKWAPFTKELAVMIVRSVNGLVFSYPIVETIHKDNICDLC
YAPARVPDSVQLKAKLLAENAIKSFPGCGIFGVEMFYLETGELLINEIAPRPHNSGHYT
IDACVTSQFEAHLRSILDLPMPKNFTSFSTITTNAIMLNVLGDKHTKDKELETCERALA
TPGSSVYLYGKESRPNRKVGHINIIASSMAECEQRLNYITGRTDIPIKISVAQKLDLEA
MVKPLVGIIMGSDSDLPVMSAACAVLKDFGVPFEVTIVSAHRTPHRMSAYAISASKRGI
KTIIAGAGGAAHLPGMVAAMTPLPVIGVPVKGSCLDGVDSLHSIVQMPRGVPVATVAIN
NSTNAALLAVRLLGAYDSSYTTKMEQFLLKQEEEVLVKAQKLETVGYEAYLENK"
rep_origin complement(6017..6605)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(6776..7636)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(7637..7741)
/gene="bla"
/note="AmpR promoter"