pCAD16_CAD_12-490_pvp008

124842 Synthetic Bacterial Expression

Plasmid Information

Purpose
Expresses Aspergillus terreus CAD in E.coli
Depositor
Konrad Buessow
Article
Chen et al Proc Natl Acad Sci U S A. 2019 Sep 23. p
Insert
cis-aconitate decarboxylase
Tags
StrepTag and TEV site
Mutation
deleted amino acids 1-11, synthetic codon optimiz…
Promoter
T7
Available Since
Sept. 16, 2019
Availability
Available

Sequence Information

Length
4,956 bp
Topology
circular
Molecule Type
other DNA
View on AddGene.org

Features

CDS 5
Label Gene Product Position Strand
Strep-Tag II — peptide that binds Strep-Tactin(R), an … 181..205 Fwd
DNA Sequence
TGGAGCCACCCGCAGTTCGAAAAA
Protein Sequence
WSHPQFEK
TEV site — tobacco etch virus (TEV) protease recog… 211..232 Fwd
DNA Sequence
GAGAACCTCTACTTCCAGGGA
Protein Sequence
ENLYFQG
S-Tag — affinity and epitope tag derived from p… 1704..1749 Fwd
DNA Sequence
AAAGAAACCGCTGCTGCGAAATTTGAACGCCAGCACATGGACTCG
Protein Sequence
KETAAAKFERQHMDS
KanR aph(3')-Ia aminoglycoside phosphotransferase 2077..2893 Rev
DNA Sequence
ATGAGCCATATTCAACGGGAAACGTCTTGCTCTAGGCCGCGATTAAATTCCAACATGGATGCTGATTTATATGGGTATAAATGGGCTCGCGATAATGTCGGGCAATCAGGTGCGACAATCTATCGATTGTATGGGAAGCCCGATGCGCCAGAGTTGTTTCTGAAACATGGCAAAGGTAGCGTTGCCAATGATGTTACAGATGAGATGGTCAGACTAAACTGGCTGACGGAATTTATGCCTCTTCCGACCATCAAGCATTTTATCCGTACTCCTGATGATGCATGGTTACTCACCACTGCGATCCCCGGGAAAACAGCATTCCAGGTATTAGAAGAATATCCTGATTCAGGTGAAAATATTGTTGATGCGCTGGCAGTGTTCCTGCGCCGGTTGCATTCGATTCCTGTTTGTAATTGTCCTTTTAACAGCGACCGCGTATTTCGTCTCGCTCAGGCGCAATCACGAATGAATAACGGTTTGGTTGATGCGAGTGATTTTGATGACGAGCGTAATGGCTGGCCTGTTGAACAAGTCTGGAAAGAAATGCATAAACTTTTGCCATTCTCACCGGATTCAGTCGTCACTCATGGTGATTTCTCACTTGATAACCTTATTTTTGACGAGGGGAAATTAATAGGTTGTATTGATGTTGGACGAGTCGGAATCGCAGACCGATACCAGGATCTTGCCATCCTATGGAACTGCCTCGGTGAGTTTTCTCCTTCATTACAGAAACGGCTTTTTCAAAAATATGGTATTGATAATCCTGATATGAATAAATTGCAGTTTCATTTGATGCTCGATGAGTTTTTCTAA
Protein Sequence
MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF
lacI lacI lac repressor 3835..4918 Rev
DNA Sequence
GTGAAACCAGTAACGTTATACGATGTCGCAGAGTATGCCGGTGTCTCTTATCAGACCGTTTCCCGCGTGGTGAACCAGGCCAGCCACGTTTCTGCGAAAACGCGGGAAAAAGTGGAAGCGGCGATGGCGGAGCTGAATTACATTCCCAACCGCGTGGCACAACAACTGGCGGGCAAACAGTCGTTGCTGATTGGCGTTGCCACCTCCAGTCTGGCCCTGCACGCGCCGTCGCAAATTGTCGCGGCGATTAAATCTCGCGCCGATCAACTGGGTGCCAGCGTGGTGGTGTCGATGGTAGAACGAAGCGGCGTCGAAGCCTGTAAAGCGGCGGTGCACAATCTTCTCGCGCAACGCGTCAGTGGGCTGATCATTAACTATCCGCTGGATGACCAGGATGCCATTGCTGTGGAAGCTGCCTGCACTAATGTTCCGGCGTTATTTCTTGATGTCTCTGACCAGACACCCATCAACAGTATTATTTTCTCCCATGAAGACGGTACGCGACTGGGCGTGGAGCATCTGGTCGCATTGGGTCACCAGCAAATCGCGCTGTTAGCGGGCCCATTAAGTTCTGTCTCGGCGCGTCTGCGTCTGGCTGGCTGGCATAAATATCTCACTCGCAATCAAATTCAGCCGATAGCGGAACGGGAAGGCGACTGGAGTGCCATGTCCGGTTTTCAACAAACCATGCAAATGCTGAATGAGGGCATCGTTCCCACTGCGATGCTGGTTGCCAACGATCAGATGGCGCTGGGCGCAATGCGCGCCATTACCGAGTCCGGGCTGCGCGTTGGTGCGGACATCTCGGTAGTGGGATACGACGATACCGAAGACAGCTCATGTTATATCCCGCCGTTAACCACCATCAAACAGGATTTTCGCCTGCTGGGGCAAACCAGCGTGGACCGCTTGCTGCAACTCTCTCAGGGCCAGGCGGTGAAGGGCAATCAGCTGTTGCCCGTCTCACTGGTGAAAAGAAAAACCACCCTGGCGCCCAATACGCAAACCGCCTCTCCCCGCGCGTTGGCCGATTCATTAATGCAGCTGGCACGACAGGTTTCCCGACTGGAAAGCGGGCAGTGA
Protein Sequence
MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ
promoter 3
Label Gene Product Position Strand
lacI promoter lacI — 0..4956 Rev
DNA Sequence
GACACCATCGAATGGCGCAAAACCTTTCGCGGTATGGCATGATAGCGCCCGGAAGAGAGTCAATTCAGGGTGGTGAAT
T7 promoter — — 85..104 Fwd
DNA Sequence
TAATACGACTCACTATAGG
AmpR promoter bla — 2893..2985 Rev
DNA Sequence
TTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGT
protein_bind 1
Label Gene Product Position Strand
lac operator — — 104..129 Fwd
DNA Sequence
GGAATTGTGAGCGGATAACAATTCC
rep_origin 1
Label Gene Product Position Strand
ColA ori — — 3002..3638 Rev
DNA Sequence
TGGTGTCGGGAATCCGTAAAGGATCTTCTTGAGATCCTTTTACGATCGTCGTAATCTCCTGCTCTGTAAACGAAAAAACCGCCTGGGGAGGCGGTTTGATCGAAGGTTAAGTCAGTTGGGGAACTGCTTAACCTGGTAACTGGCTTTAGTGGAGCGCAGATACCAAATACTGTCCTTTCAGTGTAGCCTCTGTTAGGCCACCACTTCAAGACTCTCGATATCTAAATCCACTAATTCTCAGTTACCAATGGCTGCTGCCAGTGGCGTTTTGTCGTGTCTTTCCGGGTTGGACTCAAGATGATAGTTACCGGATAAGGCGCAGCAGTCGGGCTGAACGGGGGGTTCTTGCACACAGCCCAGCTTGGAGCGAACTGTCTACACGGAACGGGACGTGGTGATTTGGGTAAAGCCTCCACCACAACACGGACGCCGCAGGACGGGAACAGGAGAGCGCAAGAGGGAGCCATCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCACTGATTTGAGCGTCAGATTTCGTGATGTTCGTCAGGGGGGCGGAGCCTATGGAAAAACGGCTTCGCTCCGGCCTTATTGTCTCTCTGCTAAGTATCCTCCTGGCATCTTCTAGGACGTTT
terminator 1
Label Gene Product Position Strand
T7 terminator — — 1800..1848 Fwd
DNA Sequence
CTAGCATAACCCCTTGGGGCCTCTAAACGGGTCTTGAGGGGTTTTTTG