pBS-SKII-3XHA-HALO-URA

188932 Synthetic Yeast Expression

Plasmid Information

Purpose
Integration cassette to create C-terminal fusions for fluorescent tagging
Depositor
Stephen Buratowski
Article
Baek et al G3 (Bethesda). 2022 Aug 9. pii: 6659097.
Insert
3XHA-HALO-URA
Available Since
Sept. 8, 2022
Availability
Available

Sequence Information

Length
5,424 bp
Topology
circular
Molecule Type
other DNA
View on AddGene.org

Features

CDS 3
Label Gene Product Position Strand
3xHA — three tandem HA epitope tags 744..834 Fwd
DNA Sequence
TACCCATACGATGTTCCTGACTATGCGGGCTATCCCTATGACGTCCCGGACTATGCAGGATCCTATCCATATGACGTTCCAGATTACGCT
Protein Sequence
YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA
HaloTag(R) — modified bacterial dehalogenase that fo… 864..1755 Fwd
DNA Sequence
ATGGCAGAAATCGGTACTGGCTTTCCATTCGACCCGCATTATGTGGAAGTCCTGGGCGAGAGAATGCATTACGTTGACGTGGGTCCGAGAGATGGAACTCCGGTCCTTTTTCTGCACGGGAATCCTACAAGCTCTTATGTTTGGCGCAATATCATCCCTCATGTAGCTCCGACGCATCGCTGTATTGCGCCGGACCTGATTGGTATGGGAAAATCTGATAAACCAGACCTGGGTTACTTTTTCGATGATCATGTGCGTTTCATGGATGCCTTCATTGAGGCATTAGGGCTTGAAGAAGTCGTCTTGGTGATTCATGATTGGGGCTCAGCTCTGGGATTTCACTGGGCTAAAAGAAATCCTGAACGCGTAAAAGGCATCGCGTTTATGGAGTTCATTCGTCCAATTCCGACTTGGGATGAATGGCCTGAGTTCGCGAGAGAAACATTTCAAGCATTTCGCACGACCGATGTAGGCCGGAAGTTAATCATCGATCAGAATGTCTTTATCGAAGGGACATTGCCGATGGGAGTCGTTCGTCCGTTAACAGAAGTCGAAATGGATCACTATAGAGAACCTTTTCTTAATCCTGTGGACAGAGAGCCGCTGTGGCGGTTTCCGAACGAACTGCCGATTGCAGGCGAGCCTGCTAACATTGTAGCGCTGGTTGAAGAGTATATGGATTGGCTTCATCAGTCTCCAGTTCCGAAGTTATTGTTTTGGGGTACGCCTGGCGTGCTTATTCCACCGGCCGAAGCGGCACGTTTGGCAAAAAGCCTGCCAAATTGCAAAGCCGTTGACATTGGCCCTGGACTTAACTTGCTTCAAGAGGATAACCCGGACTTAATCGGGAGCGAAATTGCCCGGTGGCTTTCTACCTTAGAAATCAGCGGC
Protein Sequence
MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSYVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIHDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIIDQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVEEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLIGSEIARWLSTLEISG
AmpR bla beta-lactamase 4445..5306 Rev
DNA Sequence
ATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAA
Protein Sequence
MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW
promoter 5
Label Gene Product Position Strand
T7 promoter — — 635..654 Fwd
DNA Sequence
TAATACGACTCACTATAGG
TEF promoter — — 1812..2156 Fwd
DNA Sequence
GACATGGAGGCCCAGAATACCCTCCTTGACAGTCTTGACGTGCGCAGCTCAGGGGCATGATGTGACTGTCGCCCGTACATTTAGCCCATACATCCCCATGTATAATCATTTGCATCCATACATTTTGATGGCCGCACGGCGCGAAGCAAAAATTACGGCTCCTCGCTGCAGACCTGCGAGCAGGGAAACGCTCCCCTCACAGACGCGTTGAATTGTCCCCACGCCGCGCCCCTGTAGAGAAATATAAAAGGTTAGGATTTGCCACTGAGGTTCTTCTTTCATATACTTCCTTTTAAAATCTTGCTAGGATACAGTTCTCACATCACATCCGAACATAAACAACC
T3 promoter — — 3245..3264 Rev
DNA Sequence
AATTAACCCTCACTAAAGG
lac promoter — — 3332..3363 Rev
DNA Sequence
TTTACACTTTATGCTTCCGGCTCGTATGTTG
AmpR promoter bla — 5306..5411 Rev
DNA Sequence
CGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGT
protein_bind 2
Label Gene Product Position Strand
lac operator — — 3308..3325 Fwd
DNA Sequence
TTGTTATCCGCTCACAA
CAP binding site — — 3377..3399 Fwd
DNA Sequence
ATGAGTGAGCTAACTCACATTA
rep_origin 2
Label Gene Product Position Strand
f1 ori — — 39..468 Rev
DNA Sequence
ACGCGCCCTGTAGCGGCGCATTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCGTGACCGCTACACTTGCCAGCGCCCTAGCGCCCGCTCCTTTCGCTTTCTTCCCTTCCTTTCTCGCCACGTTCGCCGGCTTTCCCCGTCAAGCTCTAAATCGGGGGCTCCCTTTAGGGTTCCGATTTAGTGCTTTACGGCACCTCGACCCCAAAAAACTTGATTAGGGTGATGGTTCACGTAGTGGGCCATCGCCCTGATAGACGGTTTTTCGCCCTTTGACGTTGGAGTCCACGTTCTTTAATAGTGGACTCTTGTTCCAAACTGGAACAACACTCAACCCTATCTCGGTCTATTCTTTTGATTTATAAGGGATTTTGCCGATTTCGGCCTATTGGTTAAAAAATGAGCTGATTTAACAAAAATTTAACGCGAATT
ori — — 3686..4275 Rev
DNA Sequence
TTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTTCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAA
terminator 1
Label Gene Product Position Strand
TEF terminator — — 2974..3172 Fwd
DNA Sequence
ACTGACAATAAAAAGATTCTTGTTTTCAAGAACTTGTCATTTGTATAGTTTTTTTATATTGTAGTTGTTCTATTTTAATCAAATGTTAGCGTGATTTATATTTTTTTTCGCCTCGACATCATCTGCCCAGATGCGAAGTTAAGTGCGCAGAAAGTAATATCATGCGTCAATCGTATGTGAATGCTGGTCGCTATACTG