pTAL-Luc is designed for analyzing enhancer sequences by assaying for expression of the firefly luciferase (luc) gene from Photinus pyralis (1). This vector contains a TATA-like promoter (PTAL) region from the Herpes simplex virus thymidine kinase (HSV-TK) promoter. Putative enhancers can be inserted into one of the MCSs. The luciferase coding sequence is followed by the SV40 late polyadenylation signal to ensure proper, efficient processing of the luc transcript in eukaryotic cells. Located upstream of the MCS is a synthetic transcription blocker (TB), which is composed of adjacent polyadenylation and transcription pause sites for reducing background transcription (2). The vector backbone also contains an f1 origin for single-stranded DNA production, a pUC origin of replication, and an ampicillin resistance gene for propagation and selection in E. coli.pTAL-Luc 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. The pTAL-Luc Vector 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.
| 载体基本信息 | |||
| 载体名称 | pTAL-Luc | 载体抗性 | Ampicillin |
|---|---|---|---|
| 载体长度 | 4956 bp | 载体类型 | Promoter enhancer reporter vector |
| 启动子 | TATA-like box | ||
pTAL-Luc 载体载体图谱
pTAL-Luc 载体载体序列
LOCUS Exported 4956 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 4956)
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..4956
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS 226..1878
/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 1919..2040
/note="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
rep_origin complement(2459..3047)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(3218..4078)
/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(4079..4183)
/gene="bla"
/note="AmpR promoter"
rep_origin 4210..4665
/direction=RIGHT
/note="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
polyA_signal 4796..4844
/note="synthetic polyadenylation signal"
misc_feature 4858..4949
/note="pause site"
/note="RNA polymerase II transcriptional pause signal from
the human alpha-2 globin gene"
