Gene:
Gene Symbol:
Gene Name:
Original Clone ID:
None
Keyword:
No ATG. Codon optimized
Species:
Human coronavirus NL63
Type:
cDNA
Vector Name:
pLenti6.3/V5-DEST
Format:
FUSION
Source:
Center for Personalized Diagnostics
Description:
None
Comments:
None
Mutation / Discrepancy:
No / No
Publications:
None
Authors:
Center for Personalized Diagnostics
HcCD00956575
Price:
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AVAILABLE
No restriction
Special MTA: None
Special MTA: None
Insert sequence: 1752nts
Open reading frame: 1 to 1752
CAGGTGTGCGGCCTGTTCAAGAATTGTACCAGAACACCCCTGAACCTGCCCCCTACTCAC
GCTCATACCTTCCTGTCTCTCAGTGACCAGTTTAAGACCACAGGCGATCTGGCAGTGCAG
ATCGGGTCCAACAACGTGTGCACCTACGAGCACGTGATTTCTTTCATGGGATTCCGCTTT
GACATCAGTATTCCAGGTTCACATAGCCTGTTCTGCACACGGGATTTTGCAATCCGTAAC
GTGCGGGGATGGCTGGGTATGGACGTGGAGTCTGCCCACGTGTGCGGCGATAACATTGGG
ACCAATGTGCCACTGCAGGTCGGATTCTCTAACGGTGTGAATTTTGTGGTCCAGACAGAG
GGATGCGTGAGTACTAACTTCGGCGACGTGATCAAGCCCGTGTGCGCCAAATCACCACCG
GGGGAACAGTTTAGACATCTGATCCCCCTGCTCAGGAAGGGACAGCCTTGGCTCATTGTG
CGGAGAAGGATCGTCCAGATGATTTCTGACTACCTGAGCAACCTGTCCGATATCCTGGTG
TTCGTGCTGTGGGCTGGCAGTCTGGAGCTCACTACCATGAGATATTTTGTGAAGATCGGC
CCCATCAAGTACTGCTATTGTGGGAACAGCGCCACATGCTACAACTCCGTGTCTAATGAA
TATTGCTGTTTCAAGCACGCCCTGGGATGTGACTACGTGTATAATCCCTACGCTTTTGAT
ATCCAGCAGTGGGGCTATGTGGGGAGTCTGTCACAGAACCACCATACTTTCTGCAACATT
CACCGCAATGAGCATGACGCATCCGGCGATGCCGTGATGACCCGATGCCTGGCCGTCCAC
GACTGTTTTGTTAAGAATGTGGATTGGACTGTGACCTACCCCTTCATCGCTAACGAAAAG
TTCATCAATGGCTGTGGGCGTAACGTGCAGGGTCACGTGGTGCGGGCCGCTCTGAAGCTC
TATAAACCATCAGTGATCCATGACATTGGCAACCCGAAGGGGGTCCGGTGCGCTGTTACT
GACGCAAAGTGGTACTGTTATGATAAACAGCCTGTGAACTCCAATGTCAAACTGCTCGAC
TACGATTATGCCACCCACGGACAGCTGGACGGTCTGTGCCTGTTCTGGAACTGTAATGTG
GATATGTACCCAGAGTTCTCAATCGTCTGCCGCTTTGACACACGCACTCGAAGCGTCTTT
AATCTGGAAGGGGTTAACGGCGGGTCCCTCTACGTGAATAAGCACGCCTTCCATACACCC
GCTTATGATAAAAGGGCATTTGTGAAGCTGAAACCAATGCCCTTCTTCTACTTCGACGAT
TCTGACTGCGATGTCGTTCAGGAGCAGGTCAACTATGTTCCTCTGCGCGCCAGCTCCTGC
GTGACTCGATGTAATATCGGAGGTGCAGTCTGTAGTAAGCACGCCAACCTGTACCAGAAA
TATGTGGAAGCTTACAATACCTTCACACAGGCAGGCTTTAACATCTGGGTGCCTCATAGC
TTCGATGTCTATAATCTGTGGCAGATCTTTATTGAGACAAACCTGCAGTCCCTCGAAAAC
ATTGCATTCAATGTGGTCAAGAAAGGCTGTTTTACTGGAGTTGACGGCGAGCTGCCCGTC
GCCGTTGTGAATGATAAGGTGTTCGTCAGATACGGCGACGTGGATAATCTGGTGTTTACC
AACAAAACCACCCTGCCTACAAACGTGGCCTTCGAACTGTTTGCTAAGAGGAAAATGGGC
CTCACCCCCCCT
Protein sequence predicted based on nucleotide sequence
QVCGLFKNCTRTPLNLPPTHAHTFLSLSDQFKTTGDLAVQIGSNNVCTYEHVISFMGFRF DISIPGSHSLFCTRDFAIRNVRGWLGMDVESAHVCGDNIGTNVPLQVGFSNGVNFVVQTE GCVSTNFGDVIKPVCAKSPPGEQFRHLIPLLRKGQPWLIVRRRIVQMISDYLSNLSDILV FVLWAGSLELTTMRYFVKIGPIKYCYCGNSATCYNSVSNEYCCFKHALGCDYVYNPYAFD IQQWGYVGSLSQNHHTFCNIHRNEHDASGDAVMTRCLAVHDCFVKNVDWTVTYPFIANEK FINGCGRNVQGHVVRAALKLYKPSVIHDIGNPKGVRCAVTDAKWYCYDKQPVNSNVKLLD YDYATHGQLDGLCLFWNCNVDMYPEFSIVCRFDTRTRSVFNLEGVNGGSLYVNKHAFHTP AYDKRAFVKLKPMPFFYFDDSDCDVVQEQVNYVPLRASSCVTRCNIGGAVCSKHANLYQK YVEAYNTFTQAGFNIWVPHSFDVYNLWQIFIETNLQSLENIAFNVVKKGCFTGVDGELPV AVVNDKVFVRYGDVDNLVFTNKTTLPTNVAFELFAKRKMGLTPP
Coding Sequence Details
Insert Sequence Verified?:
Y
Verification Method:
Sequence Verification
Mutations:
None
Discrepancy:
None
5' Linker Sequence:
None
3' Linker Sequence:
None
Recommended Growth Condition:
Distributed in bacterial strain:
DH5-alpha T1 phage resistant
Antibiotic Selection:
Host Type:
bacterial
Marker:
ampicillin
Host Type: mammalian Marker: blasticidin
Host Type: bacterial Marker: chloramphenicol
Host Type: mammalian Marker: blasticidin
Host Type: bacterial Marker: chloramphenicol
Bacterial Selection Condition:
100 ug/mL ampicillin
Growth Condition:
Growth with the single antibiotic in LB at 37 degrees is recommended.
Comments:
Commonly used conditions for ampicillin resistant plasmid clones.
Protein Expression Results:
None
Recommended expression in:
Not Applicable
Vector Name:
pLenti6.3/V5-DEST
Synonyms:
''
Sequencing Primer:
Forward:
CMV forward
Reverse:
V5 reverse
Description:
with tag/fusion/marker assay, Gateway®, acceptor (destination), recombinational cloning clone, lentiviral infection, mammalian expression expression with RSV and 5pLTR and HIV-1 psi and RRE and gp41 peptide and cPPT and CMV Promoter and CMV enhancer and attR1 and lac UV5 promoter and CmR and ccdB and attR2 and V5 and WPRE and Factor Xa site and SV40 promoter and SV40 Ori and T7 promoter and M13 forward and f1 ori and AmpR promoter and AmpR and ori and CAP binding site and lac promoter and lac operator and M13 reverse and T3 and EM7 and BlastR and 3pLTR and SV40 poly(A) signal; ampicillin resistance in bacterial, chloramphenicol resistance in bacterial, blasticidin resistance in mammalian;
Comments:
pLent6.3/V5-DEST from Invitrogen: https://www.thermofisher.com/order/catalog/product/V53306
Catalog: V53306
For research use only. Not intended for any therapeutic or diagnostic use.
The upgrade from pLenti6 to pLenti6.3 is in the addition of a WPRE cassette. Lentiviral Gateway destination vector for expression of a C-terminally V5-tagged protein.
Size (bp):
9387
Parent Vector:
None
Properties:
Gateway, acceptor (destination), lentiviral infection, mammalian expression, recombinational cloning, with tag/fusion/marker
Author Name:
Invitrogen
Publications:
None
Vector Map:
Vector Sequence:
Sequence:
Vector Features:
Type
Name
Description
Start
End
CDS
V5
epitope tag from simian virus 5
4325
4366
CDS
gp41 peptide
antigenic peptide corresponding to amino acids 655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik et al., 2013)
1493
1537
CDS
CmR
confers resistance to chloramphenicol
2801
3460
CDS
ccdB
CcdB, a bacterial toxin that poisons DNA gyrase
3802
4107
CDS
Factor Xa site
Factor Xa recognition and cleavage site
4861
4872
CDS
AmpR
confers resistance to ampicillin, carbenicillin, and related antibiotics
7302
8162
Miscellaneous Feature
WPRE
woodchuck hepatitis virus posttranscriptional regulatory element
4390
4978
Miscellaneous Feature
cPPT
central polypurine tract and central termination sequence of HIV-1
1804
1921
Miscellaneous Feature
HIV-1 psi
packaging signal of human immunodeficiency virus type 1
457
582
Miscellaneous Feature
RRE
The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm.
1075
1308
enhancer
CMV enhancer
human cytomegalovirus immediate early enhancer
1944
2247
primer_bind
M13 forward
common sequencing primer, one of multiple similar variants
6558
6574
primer_bind
M13 reverse
common sequencing primer, one of multiple similar variants
9307
9323
promoter
lac UV5 promoter
E. coli lac promoter with an "up" mutation
2717
2747
promoter
SV40 promoter
SV40 enhancer and early promoter
4993
5309
promoter
T7 promoter
promoter for bacteriophage T7 RNA polymerase
6530
6548
promoter
CMV Promoter
human cytomegalovirus (CMV) immediate early promoter
2248
2451
promoter
RSV
Rous sarcoma virus enhancer/promoter
3
229
promoter
AmpR promoter
null
7197
7301
promoter
lac promoter
promoter for the E. coli lac operon
9245
9275
promoter
T3
promoter for bacteriophage T3 RNA polymerase
9344
9362
protein binding site
attR1
recombination site for the Gateway LR reaction
2568
2692
protein binding site
attR2
recombination site for the Gateway LR reaction
4148
4272
protein binding site
CAP binding site
CAP binding activates transcription in the presence of cAMP.
9209
9230
protein binding site
lac operator
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).
9283
9299
rep_origin
SV40 Ori
SV40 origin of replication
5160
5295
rep_origin
f1 ori
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
6716
7171
rep_origin
ori
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
8333
8921
viral LTR
5pLTR
truncated 5' long terminal repeat (LTR) from HIV-1
230
410
