Gene:
Gene Symbol:
Gene Name:
Original Clone ID:
FLH162868.01X
Keyword:
None
Species:
Saccharomyces cerevisiae
Type:
cDNA
Vector Name:
pBY011
Format:
CLOSED
Source:
HIP
Description:
None
Comments:
None
Mutation / Discrepancy:
No / Yes
Publications:
None
Authors:
ScCD00094349
Price:
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AVAILABLE
No restriction
Special MTA: None
Special MTA: None
Insert sequence: 3252nts
Open reading frame: 1 to 3252
ATGTCATCTGAGGATATCATTTATGATCCTCAGTTCAAACCTGTTCAGGGAATCTACGAG
AATAGATTAAGACAATTTATTGATACTGGTGGGGATTACCATGACTTAAATTTACCCAAG
TTTTATGATAAGAAGCGCATTTCATTAGATCATGATCACGTTAAGGTTTGGTGGTACCAG
GTTTCTTTTGAACGTGGATCTTCCCCTGTTTCCCCTGATAAGAGGCCATCTTGGAAATCT
ATTATTGAACGTGACAAGAAGGGTGAATTGGAGTTTAGAGAAGCGAACATAAATCAGCCC
TTTGGACCCAGTTGGTCTACCACTTGGTTCAAGGTGAAAATCTCTTTGCCTGAAGATTGG
GTTAAATCAAACGAGCAACTACTTTTCCAATGGGATTGTTCTAACGAAGGAATTGTCATA
GACCCAAAGACTCTAATCCCTGTGACTGCTTTTTCAGGTGGTGAAAGGACTGAATATGTT
TTACCGAAAACATCGGATGGGAAACATTTCTTTTACATCGAAGCTGGTAATAATGGTATG
TTTGGCTGCGGTGCAGGATCCACTATAAATCCACCAGATGATAATAGATTTTTTCATTTG
AGAAAAGCTGATATCGTTTGGCCCGATCTAGATGCTCGTGCATTGTATATCGATTTTTGG
ATGCTAGGGGATGCTGCAAGAGAGCTACCTGGAGACTCCTGGCAAAAACACCAGGCAAGA
CAACTAGGCAATGCAGTTATGAATCTTTTTGACCCGAATGATCGTTCAAGTGTTCGTAAA
TGTCGTGAATTACTACAAAGGGAATACTTTGACTCCTTTTTAGAAAGCAGTAAGGTTTAC
GAACAAGGTGAATCTCAAGTTTTAACAAACGTTTATGGTATCGGCAACTGTCATATAGAT
ACAGCTTGGCTATGGCCATTCGCAGAAACAAGAAGGAAAATTGTGAGATCTTGGTCTTCC
CAATGTACTTTAATGGATCGCTTCCCAGAGTATAAATTTGTTGCTTCACAAGCCCAACAA
TTTAAATGGTTATTAGAAGATCATCCCGAATTTTTCAATAAAGTGTTAATTCCAAAAATT
CAGCAATCTCAGTTTTTTGCTGTTGGTGGTACATGGGTTGAGAATGATACCAATATTCCC
TCAGGGGAATCGCTGGCAAGGCAATTCTTTTTTGGTCAAAGGTTTTTCTTAAAGCATTTT
GGTCTCAAGTCAAAGATATTTTGGTTACCTGACACCTTTGGTTATTCTTCGCAAATGCCA
CAACTTTGCCGCTTATCTGGTATTGATAAATTTTTGACTCAAAAGCTTTCCTGGAATAAC
ATCAACAGTTTTCCGTATAGCACGTTTAACTGGGCAGGGATTGACGGTTCTCAATTATTG
ACTCACATGCCTCCTGGTAATACGTATACAGCAGATTCACACTTCGGTGATGTTTTGCGT
ACTGCCAAGCAGAATAAGACACCAGAATATTACGGCTCCGGTTTAATGTTGTACGGAAAG
GGTGATGGTGGTGGAGGACCAACTGAGGAAATGCTGCAAAAAATGAGGCGTATTAGATCC
ATGAATAACAGAAATGGTAATGTTATTCCAAAGTTACAAGTTGGTATAACGGTTGATGAA
TTTTATGATGACATTTTGAAAAGGACTAACCAAGGCCATGATTTACCCACATGGAGTGGT
GAATTGTATTTCGAATTTCATAGGGGGACATACACCAGTCAAGCTCAGACCAAAAAATTA
ATGAGGTTGTCCGAAATTAAACTACATGATTTGGAATGGATAGCTGCCAAAACATCAGTT
TTATATCCCGATTCGTATAAATACCCTTCCAAACAGATTAACGAACTTTGGGAGAATGTT
TTGTTATGTCAATTTCACGATGTTCTACCAGGATCATGTATTGAAATGGTATATAAGTAT
GAGGCTGTCCCTATGTTGCACAACGTTGTGAAAGAATGTACTTCCTTAATAGACAAAACC
GTTCAATTTCTTCAGAGTCAGAGTAAGGCAGATTTGGTTGAGATGAGAACATTAACTTGG
TCAAAACCAGAAAAGGTTTCTGAGGAGTGCTCGTTGAACGGTAGTTACACGTCCTCTGTA
ACCGGGTACGATGACTATATAGTCCTTGCCAATGGGAAGTTGAAAGTTATCATTTGCAAA
AAGACTGGGGTGATCACAAGCATTACGGATGAAACTTTGGGGGTAGAATACCTGGACACG
GAACATGGCAGAAACAAATTGGGCGCCAACCAGTTTGTTATTTATGATGATAAACCTTTG
GGCTGGCAGGCTTGGGATACTGAACTTTATTCCGTCAACCAATACAAGTATGTAACAAAG
CCGAAGAAAGTTCAAGTTTCCTGCAACACAAAAGAGAAATGTGCTGTTGAGGTCATTTTT
CAAATTTCCGAAAAATGCAAAATCAAATCAGTGATATCACTCAATGCAACTGCGGTCACT
GATGCTAAATTAAGTAAAGTAGACATTTCTACGACGGTAGAAAACTGGGATGCTAGAAAC
AAGTTTTTAAAAGTTGAATTTCCCGTTAATATTCGGAACGATTTTGCTTCGTATGAAACT
CAATTCGGGATTACCAAAAGACCAACACATTATAATACCTCATGGGACGTCGCTAAATTT
GAGGTTTGCCATCACAAATTTGCTGATTATTCTGAATACAGCAAGGGCGTTTCAATTTTG
AACGATTGCAAATATGGGTTTTCCACACGCGGTAACTTGATGAGATTATCATTATTAAGG
TCACCAAAGGCTCCGGATGCGCATGCTGATATGGGTACTCATGAGATAAAATATGCTATC
TATCCACATAGAGGAGCACTATCAAGCGACACTGTTAAGCTCGCTCACGAATTCAATTAT
TGCTTCAAATACAAACTCCCCAAAGATATTGGCATGAATTTTGATGACATCATAAGCATT
TCCGGAGATGAGAATGTCATTTTGTCTAACATAAAAAGAGGCGAAGATGATAGTGCTGTC
AAGTCCAACTATTCATTAAATCCAAGGGATGAACAAAGTATTGTGGTAAGGGTTTACGAA
TCTCTTGGCGGGGAATCTTTTGCTTCACTAAATACCACACTAAATTTGAAGCGTATTGAG
AAGGTCGATAACTTGGAGATGAAAGTTTATAAGAGTTTGACAGCAACGCGAGACGAATCA
AATCATGCAATAAACAGAATTCCCATTAAATTGAGACCTTTTGAGATTGCCTCATTCAGG
TTGTATTTCTGA
Protein sequence predicted based on nucleotide sequence
MSSEDIIYDPQFKPVQGIYENRLRQFIDTGGDYHDLNLPKFYDKKRISLDHDHVKVWWYQ VSFERGSSPVSPDKRPSWKSIIERDKKGELEFREANINQPFGPSWSTTWFKVKISLPEDW VKSNEQLLFQWDCSNEGIVIDPKTLIPVTAFSGGERTEYVLPKTSDGKHFFYIEAGNNGM FGCGAGSTINPPDDNRFFHLRKADIVWPDLDARALYIDFWMLGDAARELPGDSWQKHQAR QLGNAVMNLFDPNDRSSVRKCRELLQREYFDSFLESSKVYEQGESQVLTNVYGIGNCHID TAWLWPFAETRRKIVRSWSSQCTLMDRFPEYKFVASQAQQFKWLLEDHPEFFNKVLIPKI QQSQFFAVGGTWVENDTNIPSGESLARQFFFGQRFFLKHFGLKSKIFWLPDTFGYSSQMP QLCRLSGIDKFLTQKLSWNNINSFPYSTFNWAGIDGSQLLTHMPPGNTYTADSHFGDVLR TAKQNKTPEYYGSGLMLYGKGDGGGGPTEEMLQKMRRIRSMNNRNGNVIPKLQVGITVDE FYDDILKRTNQGHDLPTWSGELYFEFHRGTYTSQAQTKKLMRLSEIKLHDLEWIAAKTSV LYPDSYKYPSKQINELWENVLLCQFHDVLPGSCIEMVYKYEAVPMLHNVVKECTSLIDKT VQFLQSQSKADLVEMRTLTWSKPEKVSEECSLNGSYTSSVTGYDDYIVLANGKLKVIICK KTGVITSITDETLGVEYLDTEHGRNKLGANQFVIYDDKPLGWQAWDTELYSVNQYKYVTK PKKVQVSCNTKEKCAVEVIFQISEKCKIKSVISLNATAVTDAKLSKVDISTTVENWDARN KFLKVEFPVNIRNDFASYETQFGITKRPTHYNTSWDVAKFEVCHHKFADYSEYSKGVSIL NDCKYGFSTRGNLMRLSLLRSPKAPDAHADMGTHEIKYAIYPHRGALSSDTVKLAHEFNY CFKYKLPKDIGMNFDDIISISGDENVILSNIKRGEDDSAVKSNYSLNPRDEQSIVVRVYE SLGGESFASLNTTLNLKRIEKVDNLEMKVYKSLTATRDESNHAINRIPIKLRPFEIASFR LYF*
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
3252
Mutations:
None
Discrepancy:
t870c; g1191a;
c1336t,H446Y; a2523g;
t2688c; a2729g,H910R
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
