The pGL4.19[luc2CP /Neo] Vector(a–e) encodes the luciferase reporter gene luc2CP (Photinus pyralis) and is designed for high expression and reduced anomalous transcription. This vector also contains a mammalian selectable marker for neomycin resistance in which the number of transcription factor binding sites has been reduced and mammalian codon usage optimized. This vector is engineered with fewer consensus regulatory sequences for reduced background and a decreased risk of anomolous transcription and has a synthetic reporter gene, which has been codon optimized for mammalian expression.The pGL4.19[luc2CP /Neo] Vector is a basic vector with no promoter. However, it contains a multiple cloning region that allows cloning of a promoter of choice. The luc2CP reporter gene contains two protein destabilization sequences, hCL1 and hPEST. The protein encoded by luc2CP responds more quickly and with greater magnitude to changes in transcriptional activity than the luc2 gene, its more stable counterpart.
| 载体基本信息 | |||
| 载体名称 | pGL4.19[lucCP/Neo] | 载体抗性 | Ampicillin |
|---|---|---|---|
| 载体长度 | 5778 bp | 载体类型 | Promoter Reporter Vectors |
| 筛选标记 | Neomycin | 克隆方法 | Enzyme digestion and ligation |
pGL4.19[lucCP/Neo] 载体载体图谱
pGL4.19[lucCP/Neo] 载体载体序列
LOCUS Exported 5778 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 5778)
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..5778
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS 100..1749
/codon_start=1
/product="firefly luciferase"
/note="luciferase"
/note="synthetic luc2 version of the luciferase gene"
/translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
CDS 1756..1803
/codon_start=1
/product="non-ORF yeast peptide conferring
ubiquitin-dependent degradation"
/note="hCL1"
/note="human codon-optimized"
/translation="ACKNWFSSLSHFVIHL"
CDS 1807..1926
/codon_start=1
/product="PEST degradation sequence from mouse ornithine
decarboxylase"
/note="hPEST"
/note="human codon-optimized"
/translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV"
polyA_signal 1975..2096
/note="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
promoter 2290..2647
/note="SV40 promoter"
/note="SV40 enhancer and early promoter"
rep_origin 2498..2633
/note="SV40 ori"
/note="SV40 origin of replication"
CDS 2678..3472
/codon_start=1
/product="aminoglycoside phosphotransferase"
/note="NeoR/KanR"
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
polyA_signal 3497..3545
/note="synthetic polyadenylation signal"
rep_origin complement(3872..4460)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(4660..5520)
/codon_start=1
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
polyA_signal 5625..5673
/note="synthetic polyadenylation signal"
misc_feature 5687..5778
/note="pause site"
/note="RNA polymerase II transcriptional pause signal from
the human alpha-2 globin gene"
