pTA-Luc is a member of the MercuryTM product line of signal transduction reporter vectors. This vector is designed for analyzing enhancer sequences by assaying for expression of the firefly luciferase (luc) gene from Photinus pyralis (1). In addition, pTA-Luc contains the minimal TA promoter, the TATA box from the herpes simplex virus thymidine kinase promoter (PTA). Located downstream of PTA is the luciferase reporter gene. pTA-Luc vector maintains constitutive levels of luciferase in transformed cells.The vector backbone also contains a pUC origin of replication, and an ampicillin resistance gene for propagation and selection in E. coli.pTA-Luc is provided as a control for monitoring constitutive levels of luciferase activity in MercuryTM pathway profiling experiments. This vector is ideal for use as a negative control or for studying putative enhancers that are inserted upstream of the luciferase reporter gene. Luciferase is a highly sensitive enzymatic reporter that can be assayed by any standard luciferase-detection method, providing quantitative data on induction levels. pTA-Luc can be transfected into mammalian cells by any standard method. For selecting stable clones, cotransfect with a vector containing an antibiotic resistance gene, such as neomycin, hygromycin, or puromycin, and select resistant clones.
| 载体基本信息 | |||
| 载体名称 | pTA-Luc | 载体抗性 | Ampicillin |
|---|---|---|---|
| 载体长度 | 4871 bp | 载体类型 | Signal Pathway Reporter Vectors |
| 克隆方法 | Enzyme digestion and ligation | ||
pTA-Luc 载体载体图谱
pTA-Luc 载体载体序列
LOCUS Exported 4871 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 4871)
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..4871
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 48..79
/note="minP"
/note="minimal TATA-box promoter with low basal activity"
CDS 141..1793
/codon_start=1
/gene="luc+"
/product="firefly luciferase"
/note="luciferase"
/note="enhanced luc+ version of the luciferase gene"
/translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
polyA_signal 1834..1955
/note="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
rep_origin complement(2374..2962)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(3133..3993)
/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(3994..4098)
/gene="bla"
/note="AmpR promoter"
rep_origin 4125..4580
/direction=RIGHT
/note="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
polyA_signal 4711..4759
/note="synthetic polyadenylation signal"
misc_feature 4773..4864
/note="pause site"
/note="RNA polymerase II transcriptional pause signal from
the human alpha-2 globin gene"
