Gene:
Gene Symbol:
Gene Name:
Original Clone ID:
None
Keyword:
No ATG. Codon optimized
Species:
Human coronavirus NL63
Type:
cDNA
Vector Name:
pJFT7_n3xFlag-Halo_DC
Format:
FUSION
Source:
Center for Personalized Diagnostics
Description:
None
Comments:
None
Mutation / Discrepancy:
No / No
Publications:
None
Authors:
Center for Personalized Diagnostics
HcCD00956653
Price:
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AVAILABLE
No restriction
Special MTA: HALO
Special MTA: HALO
Insert sequence: 2514nts
Open reading frame: 1 to 2514
ACTATCACCGAGCTGCTCCAGTTCGTGACCGATCCTAGCCTGATCATTGCAAGCTCCCCC
GCCCTGGTCGACCAGAGGACAATTTGCTTTAGCGTGGCCGCCCTGTCCACAGGCCTGACT
AACCAGGTGGTCAAGCCAGGGCACTTCAATGAGGAGTTCTACAACTTCCTGAGGCTGCGC
GGCTTCTTTGATGAGGGGTCTGAACTGACCCTGAAGCATTTCTTTTTCGCCCAGAACGGC
GACGCAGCCGTGAAGGACTTTGATTTCTACCGCTATAACAAGCCCACCATCCTGGATATT
TGTCAGGCCCGGGTCACATACAAGATCGTGAGCAGATACTTCGACATCTATGAAGGCGGG
TGCATTAAGGCTTGTGAGGTTGTGGTCACTAACCTGAACAAGTCCGCCGGCTGGCCCCTG
AACAAGTTTGGGAAAGCTTCTCTGTACTATGAAAGTATCTCATATGAGGAACAGGACGCA
CTCTTCGCCCTGACCAAGCGGAATGTGCTGCCAACAATGACTCAGCTGAACCTGAAGTAC
GCAATTAGCGGAAAAGAGCGAGCACGTACCGTCGGAGGCGTGAGCCTGCTCTCTACAATG
ACCACACGCCAGTACCACCAGAAGCATCTGAAATCCATCGTGAATACCCGAAACGCCACA
GTGGTCATCGGCACTACCAAGTTTTATGGCGGGTGGAACAATATGCTCAGAACCCTGATC
GATGGAGTGGAGAACCCTATGCTGATGGGTTGGGATTACCCGAAGTGCGACCGAGCACTG
CCCAATATGATCCGTATGATTTCTGCCATGGTTCTGGGGAGTAAACACGTGAATTGCTGT
ACCGCCACAGACCGGTTCTATAGACTCGGAAACGAACTGGCTCAGGTGCTCACTGAGGTG
GTGTACAGCAATGGAGGCTTCTACTTCAAGCCCGGCGGGACAACTTCTGGCGATGCCAGT
ACCGCATACGCCAATAGTATCTTTAACATTTTCCAGGCCGTGAGCAGTAACATCAATAGG
CTGCTCTCAGTGCCTAGCGACTCCTGCAACAATGTCAACGTGCGGGATCTGCAGCGGCGG
CTGTACGACAACTGTTATCGTCTGACATCAGTGGAGGAAAGCTTTATCGACGATTACTAT
GGGTATCTGCGCAAGCATTTCTCTATGATGATTCTGAGTGACGATGGAGTGGTCTGTTAC
AACAAGGATTATGCCGAACTGGGTTACATCGCTGACATTAGCGCCTTCAAGGCAACCCTG
TACTATCAGAACAATGTCTTCATGTCTACTAGTAAGTGCTGGGTGGAGGAAGATCTGACC
AAAGGACCCCACGAGTTTTGTTCCCAGCATACTATGCAGATCGTGGACAAGGATGGAACC
TACTATCTGCCATACCCGGACCCCTCCAGGATTCTGTCTGCAGGCGTGTTCGTCGACGAC
GTGGTGAAAACAGATGCCGTGGTGCTGCTCGAACGGTACGTGAGCCTGGCCATCGACGCT
TATCCGCTCTCAAAGCACCCCAACAGCGAGTACAGAAAAGTGTTTTATGTCCTGCTCGAT
TGGGTGAAGCATCTGAACAAAAACCTGAACGAGGGCGTCCTGGAATCATTCAGCGTGACC
CTGCTCGATAACCAGGAGGACAAGTTTTGGTGCGAAGATTTCTACGCCTCCATGTATGAG
AATTCTACCATCCTGCAGGCTGCAGGACTGTGCGTGGTGTGCGGTTCCCAGACAGTGCTG
AGATGCGGCGACTGTCTGCGCAAGCCAATGCTGTGCACTAAATGTGCCTACGATCACGTG
TTTGGGACAGACCATAAGTTCATCCTGGCTATTACTCCCTACGTGTGCAACGCAAGTGGC
TGTGGCGTGAGCGATGTGAAGAAACTCTATCTGGGAGGCCTGAACTACTATTGCACAAAC
CACAAGCCTCAGCTGAGCTTTCCACTGTGCTCCGCTGGAAATATCTTCGGTCTGTATAAA
AACAGTGCAACTGGCTCACTCGACGTGGAAGTGTTTAACAGGCTGGCTACTTCTGACTGG
ACCGATGTCCGCGACTACAAGCTGGCAAATGATGTGAAAGACACACTCCGGCTGTTTGCC
GCTGAGACTATTAAGGCCAAAGAGGAAAGCGTGAAGTCAAGCTACGCTTTCGCAACCCTG
AAGGAGGTTGTGGGCCCTAAAGAACTGCTCCTGAGTTGGGAGTCAGGGAAGGTCAAACCC
CCTCTGAATAGAAACTCCGTGTTTACATGCTTCCAGATCAGCAAGGATTCCAAATTCCAG
ATCGGAGAGTTTATTTTCGAGAAAGTTGAATACGGTTCAGACACTGTGACCTATAAGAGC
ACCGTGACCACAAAACTGGTCCCAGGGATGATTTTTGTGCTGACATCTCACAATGTCCAG
CCTCTGCGCGCTCCAACTATCGCAAACCAGGAGAAGTACTCCTCTATCTACAAACTGCAT
CCGGCCTTCAATGTCAGTGACGCCTACGCTAACCTGGTGCCTTACTATCAGCTC
Protein sequence predicted based on nucleotide sequence
TITELLQFVTDPSLIIASSPALVDQRTICFSVAALSTGLTNQVVKPGHFNEEFYNFLRLR GFFDEGSELTLKHFFFAQNGDAAVKDFDFYRYNKPTILDICQARVTYKIVSRYFDIYEGG CIKACEVVVTNLNKSAGWPLNKFGKASLYYESISYEEQDALFALTKRNVLPTMTQLNLKY AISGKERARTVGGVSLLSTMTTRQYHQKHLKSIVNTRNATVVIGTTKFYGGWNNMLRTLI DGVENPMLMGWDYPKCDRALPNMIRMISAMVLGSKHVNCCTATDRFYRLGNELAQVLTEV VYSNGGFYFKPGGTTSGDASTAYANSIFNIFQAVSSNINRLLSVPSDSCNNVNVRDLQRR LYDNCYRLTSVEESFIDDYYGYLRKHFSMMILSDDGVVCYNKDYAELGYIADISAFKATL YYQNNVFMSTSKCWVEEDLTKGPHEFCSQHTMQIVDKDGTYYLPYPDPSRILSAGVFVDD VVKTDAVVLLERYVSLAIDAYPLSKHPNSEYRKVFYVLLDWVKHLNKNLNEGVLESFSVT LLDNQEDKFWCEDFYASMYENSTILQAAGLCVVCGSQTVLRCGDCLRKPMLCTKCAYDHV FGTDHKFILAITPYVCNASGCGVSDVKKLYLGGLNYYCTNHKPQLSFPLCSAGNIFGLYK NSATGSLDVEVFNRLATSDWTDVRDYKLANDVKDTLRLFAAETIKAKEESVKSSYAFATL KEVVGPKELLLSWESGKVKPPLNRNSVFTCFQISKDSKFQIGEFIFEKVEYGSDTVTYKS TVTTKLVPGMIFVLTSHNVQPLRAPTIANQEKYSSIYKLHPAFNVSDAYANLVPYYQL
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
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:
pJFT7_n3xFlag-Halo_DC
Synonyms:
''
Description:
In vitro expression vector with a T7 promoter and N-terminal TEV-cleavable 3xFlag-Halo tag; ampicillin resistance in bacteria; Gateway or Flexi cloning compatible
Comments:
None
Size (bp):
6286
Parent Vector:
None
Properties:
Gateway, acceptor (destination), blue/white selection, in vitro transcription, recombinational cloning
Author Name:
Arizona State University
Joshua LaBaer
Justin Saul
Ji Qiu
Fernanda Festa
Center for Personalized Diagnostics
Joshua LaBaer
Justin Saul
Ji Qiu
Fernanda Festa
Center for Personalized Diagnostics
Publications:
None
Vector Map:
Vector Sequence:
Sequence:
Vector Features:
Type
Name
Description
Start
End
bacterial origin
ColE
ColE1 bacterial origin
3571
4253
bacterial origin
M13 Ori
M13 origin
5647
6102
death cassette
ccdB
Gateway ccdB death Cassette (DC) (removed in plasmids with inserts)
1558
3212
death cassette 2
ccdB (BsrGI removed)
ccdb(bsrgi removed)
2767
3072
enhancer
CITE
CITE enhancer
17
515
primer
NAP150
NAP150F forward sequencing primer cccattgtatgggatctgatc (sequencing starts before the 3xFlag-HALO tag)
338
408
primer
reverse primer
cHaloseqR reverse sequencing primer CAACATCGACGTAGTGCAT
657
675
primer
forward primer
HaloseqF aagcctgcctaactgcaa (used for sequencing the gene insert)
1364
1381
primer
NAP138
NAP138r reverse sequencing primer tgtttcgccatttatcacctt
3272
3292
primer
T7 term primer
T7 terminator primer
3331
3349
primer
M13 forward
M13 forward sequencing primer
6246
6263
promoter
T7
T7 promoter
6270
6286
protease cleavage site
TEV
TEV protease cleavage site EDLYFQS (used to separate expressed protein from 3xFlag-HALO tag)
1497
1517
recombination site
attR1
attR1 recombinational cloning site
1533
1557
recombination site
attR2
attR2 recombinational cloning site
3213
3237
reporter gene
LacZ
LacZ alpha
6107
6175
selectable marker
CmR
Chloramphenicol resistance gene (removed in plasmids containing gene inserts)
1766
2422
selectable marker
AmpR
Ampicillin resistance gene
4351
5010
ssDNA origin
f1
f1 origin
5779
6085
tag
Flag
N-terminal 3xFlag tag
546
590
tag
HALO
N-terminal HALO tag
597
1484
trxn termination sequence
T7 term
T7 terminator
3342
3389
