pst39-NST-clone14

133421 Saccharomyces cerevisiae Bacterial Expression

Plasmid Information

Purpose
S. cerevisiae Nip1 (with C-terminal TEV cleavage site and strep-II tag), Sui1 and Tif5B6 (with N-terminal 7xHis tag and TEV cleavage set) coding sequences with intervening ribosome binding sites and other cis-elements for expression as a polycistron, T7 theta 10 promoter (and corresponding terminator after polycistron).
Depositor
Gerhard Wagner
Insert
NIP1/SUI1/TIF5 (NIP1, TIF5, SUI1 Budding Yeast)
Mutation
ORF for Nip1 encodes aa 4-156 (not complete ORF),…
Available Since
Jan. 6, 2020
Availability
Available

Sequence Information

Length
4,359 bp
Topology
circular
Molecule Type
other DNA
View on AddGene.org

Features

CDS 5
Label Gene Product Position Strand
TEV site — tobacco etch virus (TEV) protease recog… 751..772 Fwd
DNA Sequence
GAAAACCTGTATTTTCAGGGC
Protein Sequence
ENLYFQG
Strep-Tag II — peptide that binds Strep-Tactin(R), an … 772..796 Fwd
DNA Sequence
TGGAGCCACCCGCAGTTCGAAAAA
Protein Sequence
WSHPQFEK
7xHis — 6xHis affinity tag 1322..1343 Fwd
DNA Sequence
CATCACCATCACCATCACCAT
Protein Sequence
HHHHHHH
TEV site — tobacco etch virus (TEV) protease recog… 1343..1364 Fwd
DNA Sequence
GAAAACCTGTATTTTCAGGGC
Protein Sequence
ENLYFQG
rop rop Rop protein, which maintains plasmids a… 4137..4329 Rev
DNA Sequence
GTGACCAAACAGGAAAAAACCGCCCTTAACATGGCCCGCTTTATCAGAAGCCAGACATTAACGCTTCTGGAGAAACTCAACGAGCTGGACGCGGATGAACAGGCAGACATCTGTGAATCGCTTCACGACCACGCTGATGAGCTTTACCGCAGCTGCCTCGCGCGTTTCGGTGATGACGGTGAAAACCTCTGA
Protein Sequence
MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHADELYRSCLARFGDDGENL
RBS 1
Label Gene Product Position Strand
Shine-Dalgarno sequence — — 1765..1774 Fwd
DNA Sequence
TAAGGAGGT
promoter 2
Label Gene Product Position Strand
T7 promoter — — 206..225 Fwd
DNA Sequence
TAATACGACTCACTATAGG
AmpR promoter bla — 2096..2188 Fwd
DNA Sequence
TTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGT
rep_origin 1
Label Gene Product Position Strand
ori — — 3122..3711 Fwd
DNA Sequence
TTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTCCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAA
terminator 2
Label Gene Product Position Strand
T7 terminator — — 1982..2030 Fwd
DNA Sequence
CTAGCATAACCCCTTGGGGCCTCTAAACGGGTCTTGAGGGGTTTTTTG
fd terminator — — 3052..3092 Fwd
DNA Sequence
GATACAATTAAAGGCTCCTTTTGGAGCCTTTTTTTTTGGA