Gene:
Gene Symbol:
Gene Name:
Original Clone ID:
FLH260539.01X
Keyword:
None
Species:
Saccharomyces cerevisiae
Type:
cDNA
Vector Name:
pBY011
Format:
CLOSED
Source:
HIP
Description:
None
Comments:
None
Mutation / Discrepancy:
No / No
Publications:
None
Authors:
ScCD00097878
Price:
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AVAILABLE
No restriction
Special MTA: None
Special MTA: None
Insert sequence: 2238nts
Open reading frame: 1 to 2238
ATGTCACACACTAATAAGATCGCATACGTGTTGAACAATGACACGGAGGAAACAGCCTCG
CCCTCTTCCGTTGGTTGTTTTGACAAGAAACAGCTCACTAAATTACTGATACATACTCTA
AAGGAGCTGGGCTACGACTCCGCCGCTAACCAGTTACTGCTAGAGAGCGGCGGATATCAA
AATGAGTCTAACCACATCCAGACTTTCTTCAAACTCATCAAGACCGGCCAATTCCATCTT
ATTAATTGGCAGATTGTTTGCTCGCTGCCCCTTGCCCATAGCTCTCCGCTAAGGTCCGAA
TGGCTTCAAAGGCTGCTCATTCCCACGCCGACGCCCGCCACGACTTCACTTTTCGACCAC
ATGCTCCTGCAACTGCAATACCTGCAACAGCTGATGAGCTCTGTAAATTCATCTACTTGT
TCGGACGCAGAGATCGCCACGCTCAGAAATTACGTAGAGATCATGATTCTAGTTAATAGA
CAAATATTCCTCGAATTCTTCCATCCAGTCACAAATTCCGCCTCTCACAAGGGCCCTCAC
ACTGCACTGCCCGTCCTGTATTTGCGCAAAATCCTCAAAAACTTCATCGAGATATGGGAT
TCCCTGCTGGTGTCTAACGATCAGTTCCTAAATGAAGAAAACATCTTCAATCCGGAAACG
ACCTTGAGAGAACTGTCGACGTACCTGACCAACCCAAAACTAACCGCGCAGTTGAACCTC
GAGCGAGACCATCTGATTGACGCCATCTCCAAATATATCGATCCGAACGAACTCGTTCCC
AAGGGTCGTCTCTTGCATCTCTTGAAACAGGCCATCAAGTATCAACAATCACAAGACATT
TTTAATATCATCGATCCGGATGATGACGCCTCTTTCTCCTCTCCACCCCACCGGATCAAC
CTTTTGCAGGATAACTTCTCACACGATCTGACTGTGACCTTTCAAGAATGGAAGACTATC
CAAGACACCACAGACGAAATTTGGTTTTTGACATTTTCGCCCAATGGCAAGTATTTGGCT
TCTGCGACTTCCGAGTCTTCAAGAGGCTACTTCATTACTGTTTATGATGTGGAACAAGAC
TTCAAAATTTATAAGACTTGCGTGAGCTTGTCACAATCCGTATTGTACCTCATGTTTTCT
CCTGATAGCCGGTATCTGGTTGCTTGCCCTTTCAGCGAAGATGTTACCATTTACGACATG
AATGCCACCTCCCTGCCCGATGCGTCTGCCACAGACTCGTTCCTACTATACCCTTCTACC
AGGCTCTCGCCCATGGACTCGTTTAAACTGGACACCACCACGTACCCAGATGATACCGAA
TCATCAGCATCATCTTCCTCGAGGCCGGCAAACGCGAATTCTAATCAATCAAGAGTATGG
TGCTGCGATGCCTTCCACACTGCGGAACGTGCGGGCTGGATGGTGGTTGGATCGCCCGAC
AGAGAAGCTATTGTACACTCACTCACCACGAAAGAGTCACTTTTTAGCTTAAAAGGCAGG
ACTTGTATCGCGTTGGGCCACGATGAAAACATCTCTGGGAGAAAATCAATCGATCCTGCA
AAAGTCCTTTACAAACCAACAAGTAGCAATGGTAACTGGCAATATGTGGAGGACGACGAA
ACCTTTCCACGTGTGCACGACGTGAAAATCAGCTATGACGATAAATACGTGCTTTTAATG
ACCCACCAGGGCGTTATAGATGTCTACGATTTCAGTGGGTTCCCCTCCAAAGAGGAGCTA
TCTAAGCAAACAGTGGATCCCAAAAACTTTCTGATTCCAAGGATCGCAAGATTGGACGTG
GGCAAAAACATGACTTGTATCTCATTACCGCTAAATACCACGCATCAAGGCTTCCATAGG
CAACAAATATCCGAGTCTCAACATTTGGTATTAGTTAGCCTTCAAGATAACGAATTACAA
ATGTGGGACTATAAAGAAAATATTCTCATACAAAAGTACTTTGGACAGAAGCAACAGCAC
TTTATTATCAGATCTTGCTTCGCCTACGGTAATAAGCTTGTCATGAGCGGATCAGAAGAT
GGTAAAATTTACATTTGGGATAGAATTAGAGGTAACCTAGTATCTGTTTTATCCGGTCAT
TCCACGGTAATGAGTAATTCCACAAAACCCATGGGGAAAAATTGTAACGTAGTTGCATCA
AACCCTGCAGATAAAGAAATGTTTGCTTCTGGCGGTGATGACGGTAAGATAAAAATCTGG
AAAATTTCAAGAAATTAG
Protein sequence predicted based on nucleotide sequence
MSHTNKIAYVLNNDTEETASPSSVGCFDKKQLTKLLIHTLKELGYDSAANQLLLESGGYQ NESNHIQTFFKLIKTGQFHLINWQIVCSLPLAHSSPLRSEWLQRLLIPTPTPATTSLFDH MLLQLQYLQQLMSSVNSSTCSDAEIATLRNYVEIMILVNRQIFLEFFHPVTNSASHKGPH TALPVLYLRKILKNFIEIWDSLLVSNDQFLNEENIFNPETTLRELSTYLTNPKLTAQLNL ERDHLIDAISKYIDPNELVPKGRLLHLLKQAIKYQQSQDIFNIIDPDDDASFSSPPHRIN LLQDNFSHDLTVTFQEWKTIQDTTDEIWFLTFSPNGKYLASATSESSRGYFITVYDVEQD FKIYKTCVSLSQSVLYLMFSPDSRYLVACPFSEDVTIYDMNATSLPDASATDSFLLYPST RLSPMDSFKLDTTTYPDDTESSASSSSRPANANSNQSRVWCCDAFHTAERAGWMVVGSPD REAIVHSLTTKESLFSLKGRTCIALGHDENISGRKSIDPAKVLYKPTSSNGNWQYVEDDE TFPRVHDVKISYDDKYVLLMTHQGVIDVYDFSGFPSKEELSKQTVDPKNFLIPRIARLDV GKNMTCISLPLNTTHQGFHRQQISESQHLVLVSLQDNELQMWDYKENILIQKYFGQKQQH FIIRSCFAYGNKLVMSGSEDGKIYIWDRIRGNLVSVLSGHSTVMSNSTKPMGKNCNVVAS NPADKEMFASGGDDGKIKIWKISRN*
Coding Sequence Details
Insert Sequence Verified?:
N
Verification Method:
Insert was fully sequenced in parent vector
Reference Sequence Annotations:
Start on reference sequence
1
End on reference sequence
2238
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:
yeast
Marker:
URA
Host Type: bacterial Marker: ampicillin
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:
pBY011
Synonyms:
''
Description:
with tag/fusion/marker assay, Gateway®, acceptor (destination), recombinational cloning clone, inducible expression, yeast expression, yeast shuttle expression with ARAC5R and Ura3 promoter and AmpR promoter and bla and ori and CAP binding site and lac promoter and lac operator and M13 reverse and attR2 and CmR and lac UV5 promoter and attR1 and GAL1, 10 and Ura3 and UAS and M13 forward; 50ug/mL of Kanamycin resistance in Pichia;
Comments:
This is a bacteria-yeast shuttle vector useful for expressing cDNAs in yeast that includes an auxotrophic marker for selection in yeast.
Size (bp):
8121
Parent Vector:
Name:
YCplac33
Description: Parent vector is a CEN4, URA3, ampR, ColE1 vector from R.D. Geitz described in Gene 74(2) 5270534 (1988). The Gal1-10 promoter and the Gateway-type attR recombination cassette (reference frame A, see information at the Invitrogen website) were inserted into the parent to create pBY011.
Properties:
Gateway, acceptor (destination), inducible expression, recombinational cloning, with tag/fusion/marker, yeast expression, yeast shuttle
Author Name:
Bhullar
Publications:
None
Vector Map:
Vector Sequence:
Sequence:
Vector Features:
Type
Name
Description
Start
End
CDS
CmR
confers resistance to chloramphenicol
5563
6222
CDS
Ura3
yeast auxotrophic marker, counterselectable with 5-fluoroorotic acid (5-FOA)
1460
2263
CDS
bla
allows cytosolic expression of beta-lactamase
2768
3556
primer_bind
M13 forward
common sequencing primer, one of multiple similar variants
7183
7199
primer_bind
M13 reverse
common sequencing primer, one of multiple similar variants
4702
4718
promoter
AmpR promoter
null
2590
2694
promoter
GAL1, 10
divergent inducible promoter, regulated by Gal4
6481
7145
promoter
Ura3 promoter
S. cerevisiae URA3
2264
2479
promoter
lac promoter
promoter for the E. coli lac operon
4640
4670
promoter
lac UV5 promoter
E. coli lac promoter with an "up" mutation
6276
6306
protein binding site
attR1
recombination site for the Gateway LR reaction
6331
6455
protein binding site
attR2
recombination site for the Gateway LR reaction
4747
4871
protein binding site
CAP binding site
CAP binding activates transcription in the presence of cAMP.
4604
4625
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).
4678
4694
protein binding site
UAS
upstream activating sequence mediating Gal4-dependent induction
6812
6929
rep_origin
ARAC5R
S. cerevisiae autonomously replicating sequence ARS1/ARS416
288
1125
rep_origin
ori
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
3727
4315
