pTK-Hyg is a selection vector that confers hygromycin resistance in mammalian cells for the selection of stably transformed cells. pTK-Hyg is especially useful for selection of double-stable cell lines using the Tet-On or Tet-Off Gene Expression Systems because the absence of an enhancer element on pTK-Hyg prevents the unwanted activation of pTRE- and pBI-derived plasmids upon cointegration into the host cells genome.pTK-Hyg can be cotransfected into the host cell line with an expression plasmid that contains a gene of interest using any standard transfection technique. Most mammalian cell lines require 200 μg/ml of Hygromycin to select for stably transformed cells.
| 载体基本信息 | |||
| 载体名称 | pTK-Hyg | 载体抗性 | Ampicillin |
|---|---|---|---|
| 载体长度 | 5066 bp | 载体类型 | Tetracycline Regulatory System |
| 筛选标记 | Hyg | 克隆方法 | Enzyme digestion and ligation |
pTK-Hyg 载体载体图谱
pTK-Hyg 载体载体序列
LOCUS Exported 5066 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 5066)
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..5066
/organism="synthetic DNA construct"
/mol_type="other DNA"
polyA_signal 29..163
/note="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
polyA_signal 1450..1498
/note="HSV TK poly(A) signal"
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
CDS complement(1540..2577)
/codon_start=1
/product="hygromycin B phosphotransferase"
/note="HygR"
/note="confers resistance to hygromycin"
/translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRGDRE
MGEAN"
promoter complement(2612..2757)
/note="HSV TK promoter"
/note="herpes simplex virus thymidine kinase promoter"
rep_origin complement(3214..3802)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(3973..4833)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(4834..4938)
/gene="bla"
/note="AmpR promoter"
