pLVX-PAmCherry-C1 Vector is an HIV-1-based, lentiviral expression vector. Lentiviral particles derived from the vector allow you to infect cells and express your gene of interest fused to PAmCherry, a photoactivatable mutant of the fluorescent protein mCherry. PAmCherry is non-fluorescence until photoactivated by a short exposure to light at a wavelength between 350 nm and 400 nm. The excitation/emission wavelengths of photoactivated PAmCherry are 564 nm and 595 nm. Genes cloned into the MCS will be expressed as fusions to the C-terminus of PAmCherry if they are in the same reading frame as PAmCherry and there are no intervening stop codons.
| 载体基本信息 | |||
| 载体名称 | pLVX-PAmCherry-C1 | 载体抗性 | Ampicillin |
|---|---|---|---|
| 载体长度 | 8770 bp | 载体类型 | Viral Expression & Packaging Vectors |
| 载体来源 | Clontech | 筛选标记 | Puromycin |
| 拷贝数 | High copy number | 启动子 | CMV |
| 克隆方法 | Enzyme Cut | 载体标签 | PAmCherry |
| 表达方法 | Constiutive, Stable | ||
pLVX-PAmCherry-C1 载体载体图谱
pLVX-PAmCherry-C1 载体载体序列
LOCUS Exported 8770 bp ds-DNA circular SYN 08-APR-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pLVX-PAmCherry-C1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8770)
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..8770
/organism="synthetic DNA construct"
/mol_type="other DNA"
LTR 1..634
/note="3 LTR"
/note="3 long terminal repeat (LTR) from HIV-1"
misc_feature 681..806
/note="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1303..1536
/note="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
misc_feature 2028..2143
/note="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1 (lacking the first T)"
enhancer 2201..2504
/note="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 2505..2708
/note="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
CDS 2806..3513
/codon_start=1
/product="photoactivatable monomeric derivative of DsRed
fluorescent protein"
/note="PAmCherry"
/note="mammalian codon-optimized"
/translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHVFEIEGEGEGRPYEG
TQTAKLKVTKGGPLPFTWDILSPQFMYGSNAYVKHPADIPDYFKLSFPEGFKWERVMKF
EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEALSERMYPEDGALK
GEVKPRVKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNRKLDITSHNEDYTIVEQYERA
EGRHSTGGMDELYK"
misc_feature 3526..3582
/note="MCS"
/note="multiple cloning site"
promoter 3605..4104
/note="PGK promoter"
/note="mouse phosphoglycerate kinase 1 promoter"
CDS 4125..4724
/codon_start=1
/gene="pac from Streptomyces alboniger"
/product="puromycin N-acetyltransferase"
/note="PuroR"
/note="confers resistance to puromycin"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
misc_feature 4738..5326
/note="WPRE"
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
CDS complement(5209..5220)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/note="Factor Xa site"
/translation="IEGR"
LTR 5533..6166
/note="3 LTR"
/note="3 long terminal repeat (LTR) from HIV-1"
primer_bind complement(6295..6311)
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 6319..6335
/bound_moiety="lac repressor encoded by lacI"
/note="lac operator"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(6343..6373)
/note="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 6388..6409
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(6697..7285)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(7456..8316)
/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(8317..8421)
/gene="bla"
/note="AmpR promoter"
polyA_signal 8469..8603
/note="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
