pGWKS12

139419 Synthetic Yeast Expression

Plasmid Information

Purpose
Fluoroescent marker for use in Cryptococcus neoformans. Contains a codon optimised Superfolder GFP marker.
Depositor
James Fraser
Article
Spencer et al Fungal Genet Biol. 2020 May;138:103365.
Insert
Superfolder GFP
Promoter
TEF1
Available Since
May 4, 2020
Availability
Available

Sequence Information

Length
8,406 bp
Topology
circular
Molecule Type
other DNA

Features

CDS 3
Label Gene Product Position Strand
NrsR Streptomyces noursei nat1 nourseothricin acetyltransferase 1024..1588 Rev
DNA Sequence
ACTCTTGACGACACGGCTTACCGGTACCGCACCAGTGTCCCGGGGGACGCCGAGGCCATCGAGGCACTGGATGGGTCCTTCACCACCGACACCGTCTTCCGCGTCACCGCCACCGGGGACGGCTTCACCCTGCGGGAGGTGCCGGTGGACCCGCCCCTGACCAAGGTGTTCCCCGACGACGAATCGGACGACGAATCGGACGACGGGGAGGACGGCGACCCGGACTCCCGGACGTTCGTCGCGTACGGGGACGACGGCGACCTGGCGGGCTTCGTGGTCGTCTCGTACTCCGGCTGGAACCGCCGGCTGACCGTCGAGGACATCGAGGTCGCCCCGGAGCACCGGGGGCACGGGGTCGGGCGCGCGTTGATGGGGCTCGCGACGGAGTTCGCCCGCGAGCGGGGCGCCGGGCACCTCTGGCTGGAGGTCACCAACGTTAACGCACCGGCTATCCACGCGTACCGGCGGATGGGGTTCACCCTCTGCGGCCTGGACACCGCCCTGTACGACGGCACCGCCTCGGACGGCGAGCAGGCGCTCTACATGAGCATGCCCTGCCCCTAA
Protein Sequence
TLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGFTLREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNRRLTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFTLCGLDTALYDGTASDGEQALYMSMPCP
superfolder GFP GFP variant that folds robustly even wh… 4006..4723 Fwd
DNA Sequence
ATGTCCAAGGGTGAGGAGCTTTTCACTGGTGTTGTCCCTATTCTTGTTGAGTTGGACGGTGACGTTAACGGCCACAAGTTCTCTGTCCGTGGTGAGGGTGAGGGTGACGCTACTAACGGTAAGCTCACCCTTAAGTTCATTTGCACTACTGGTAAGCTTCCTGTTCCTTGGCCTACTCTTGTCACTACTCTTACCTACGGTGTTCAGTGCTTCTCCCGTTACCCCGACCACATGAAGCGACACGACTTCTTCAAGTCCGCCATGCCCGAGGGTTACGTTCAGGAGCGTACTATCTCTTTCAAGGACGACGGCACCTACAAGACCCGTGCTGAGGTCAAGTTCGAGGGTGACACCCTTGTTAACCGTATCGAGTTGAAGGGTATTGACTTCAAGGAGGACGGTAACATTCTCGGTCACAAGCTCGAGTACAACTTCAACTCTCACAACGTTTACATCACCGCTGACAAGCAGAAGAACGGTATCAAGGCTAACTTCAAGATTCGTCACAACGTTGAGGACGGTTCCGTTCAGCTTGCTGACCACTACCAGCAGAACACTCCTATTGGCGACGGCCCTGTCCTTTTGCCTGACAACCACTACCTTTCCACTCAGTCTGTCCTTTCCAAGGACCCCAACGAGAAGCGTGACCACATGGTCCTTCTTGAGTTCGTTACTGCTGCTGGCATTACTCACGGCATGGACGAGCTCTACAAGTAG
Protein Sequence
MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDGTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLEFVTAAGITHGMDELYK
AmpR bla beta-lactamase 6612..7473 Rev
DNA Sequence
ATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAA
Protein Sequence
MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW
promoter 4
Label Gene Product Position Strand
T7 promoter 2736..2755 Fwd
DNA Sequence
TAATACGACTCACTATAGG
T3 promoter 5412..5431 Rev
DNA Sequence
AATTAACCCTCACTAAAGG
lac promoter 5499..5530 Rev
DNA Sequence
TTTACACTTTATGCTTCCGGCTCGTATGTTG
AmpR promoter bla 7473..7578 Rev
DNA Sequence
CGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGT
protein_bind 2
Label Gene Product Position Strand
lac operator 5475..5492 Fwd
DNA Sequence
TTGTTATCCGCTCACAA
CAP binding site 5544..5566 Fwd
DNA Sequence
ATGAGTGAGCTAACTCACATTA
rep_origin 1
Label Gene Product Position Strand
ori 5853..6442 Rev
DNA Sequence
TTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTTCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAA