pCDH-CMV-MCS-EF1-CopGFP-T2A-Puro
Search name
pCDH-CMV-MCS-EF1-CopGFP-T2A-Puro,Plasmid pCDH-CMV-MCS-EF1-CopGFP-T2A-Puro,pCDH-CMV-MCS-EF1-CopGFP-T2A-Puro vector
pCDH-CMV-MCS-EF1-CopGFP-T2A-Puro Information
Promoter: EF-1 alpha core, CMV
Replicator: pUC ori, SV40 ori
Terminator: SV40 poly (A) signal
Plasmid classification: virus series, lentivirus cloning vector
Plasmid size: 8189bp
Prokaryotic resistance: Amp
Screening markers: Puro
Cloned strain: Stbl3
Culture conditions: 37 centigrade, aerobic LB
Expression host: mammalian cells
Induction mode: no induction, instantaneous expression
5'sequencing primers: CMV-F:CGCAAATGGGCGGTAGGCGTG
3'sequencing primers: primers designed according to sequence
Use:Lentiviral
pCDH-CMV-MCS-EF1-CopGFP-T2A-Puro Description
PCDH-CMV-MCS-EF1-CopGFP-T2A-Puro polyclonal site (MCS) - in MCS is located in the CMV promoter downstream of interest gene cloning.To improve stability and CMV driven transcription translation. SV40 multi - poly adenosine acidification signal to enable the efficiency of transcription and transcription termination. The hybrid / RSV 5ltr promoter provides a high level of viral transcript expression in 293 cells of this full-length producer.
pCDH-CMV-MCS-EF1-CopGFP-T2A-Puro Sequence
LOCUS Exported 8189 bp ds-DNA circular SYN 30-AUG-2016
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS Untitled 2
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8189)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Tuesday, August 30, 2016 from SnapGene Viewer 3.1.4
FEATURES Location/Qualifiers
source 1..8189
/organism="synthetic DNA construct"
/mol_type="other DNA"
LTR 234..414
/note="5' LTR (truncated)"
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 458..583
/note="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1076..1309
/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 1798..1915
/note="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 2005..2208
/note="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
promoter 2348..2559
/note="EF-1-alpha core promoter"
/note="core promoter for human elongation factor
EF-1-alpha"
LTR 2572..2840
/note="5' LTR (truncated)"
/note="truncated 5' long terminal repeat (LTR) from human
T-cell leukemia virus (HTLV) type 1"
CDS 2899..3561
/codon_start=1
/product="green fluorescent protein 2 from Pontellina
plumata, also known as ppluGFP2 (Shagin et al., 2004)"
/note="CopGFP"
/translation="PAMEIECRITGTLNGVEFELVGGGEGTPKQGRMTNKMKSTKGALT
FSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVLHVSFSYRY
EAGRVIGDFKVVGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNVLVGSFARTFSLRDGG
YYSFVVDSHMHFKSAIHPSILQNGGPMFAFRRVEELHSNTELGIVEYQHAFKTPIAFA"
CDS 3631..3684
/codon_start=1
/product="2A peptide from Thosea asigna virus capsid
protein"
/note="T2A"
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="EGRGSLLTCGDVEENPGP"
CDS 3685..4284
/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 4285..4872
/note="WPRE"
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
CDS complement(4756..4767)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/note="Factor Xa site"
/translation="IEGR"
LTR 4946..5179
/note="3' LTR (Delta-U3)"
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 5251..5372
/note="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
rep_origin 5391..5526
/note="SV40 ori"
/note="SV40 origin of replication"
primer_bind complement(5564..5580)
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 5588..5604
/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(5612..5642)
/note="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 5657..5678
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(5966..6554)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"< <
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