pet16b-Avi-ubiquitin_rbs_BirA

134897 Homo sapiens Bacterial Expression

Plasmid Information

Purpose
Expresses HIS-Avi-tagged ubiquitin for recombinant protein production
Depositor
Andrew Deans
Article
Tan et al PLoS One. 2020 Feb 24;15(2):e0229000. do
Tags
10HIS-AviTag-PreScission(3C)protease
Promoter
RBS and T7
Available Since
April 14, 2020
Availability
Available

Sequence Information

Length
6,982 bp
Topology
circular
Molecule Type
other DNA

Features

CDS 8
Label Gene Product Position Strand
AmpR bla beta-lactamase 208..1069 Fwd
DNA Sequence
ATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTGTTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGCAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAA
Protein Sequence
MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW
rop rop Rop protein, which maintains plasmids a… 2254..2446 Rev
DNA Sequence
GTGACCAAACAGGAAAAAACCGCCCTTAACATGGCCCGCTTTATCAGAAGCCAGACATTAACGCTTCTGGAGAAACTCAACGAGCTGGACGCGGATGAACAGGCAGACATCTGTGAATCGCTTCACGACCACGCTGATGAGCTTTACCGCAGCTGCCTCGCGCGTTTCGGTGATGACGGTGAAAACCTCTGA
Protein Sequence
MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHADELYRSCLARFGDDGENL
lacI lacI lac repressor 3757..4840 Rev
DNA Sequence
GTGAAACCAGTAACGTTATACGATGTCGCAGAGTATGCCGGTGTCTCTTATCAGACCGTTTCCCGCGTGGTGAACCAGGCCAGCCACGTTTCTGCGAAAACGCGGGAAAAAGTGGAAGCGGCGATGGCGGAGCTGAATTACATTCCCAACCGCGTGGCACAACAACTGGCGGGCAAACAGTCGTTGCTGATTGGCGTTGCCACCTCCAGTCTGGCCCTGCACGCGCCGTCGCAAATTGTCGCGGCGATTAAATCTCGCGCCGATCAACTGGGTGCCAGCGTGGTGGTGTCGATGGTAGAACGAAGCGGCGTCGAAGCCTGTAAAGCGGCGGTGCACAATCTTCTCGCGCAACGCGTCAGTGGGCTGATCATTAACTATCCGCTGGATGACCAGGATGCCATTGCTGTGGAAGCTGCCTGCACTAATGTTCCGGCGTTATTTCTTGATGTCTCTGACCAGACACCCATCAACAGTATTATTTTCTCCCATGAAGACGGTACGCGACTGGGCGTGGAGCATCTGGTCGCATTGGGTCACCAGCAAATCGCGCTGTTAGCGGGCCCATTAAGTTCTGTCTCGGCGCGTCTGCGTCTGGCTGGCTGGCATAAATATCTCACTCGCAATCAAATTCAGCCGATAGCGGAACGGGAAGGCGACTGGAGTGCCATGTCCGGTTTTCAACAAACCATGCAAATGCTGAATGAGGGCATCGTTCCCACTGCGATGCTGGTTGCCAACGATCAGATGGCGCTGGGCGCAATGCGCGCCATTACCGAGTCCGGGCTGCGCGTTGGTGCGGATATCTCGGTAGTGGGATACGACGATACCGAAGACAGCTCATGTTATATCCCGCCGTTAACCACCATCAAACAGGATTTTCGCCTGCTGGGGCAAACCAGCGTGGACCGCTTGCTGCAACTCTCTCAGGGCCAGGCGGTGAAGGGCAATCAGCTGTTGCCCGTCTCACTGGTGAAAAGAAAAACCACCCTGGCGCCCAATACGCAAACCGCCTCTCCCCGCGCGTTGGCCGATTCATTAATGCAGCTGGCACGACAGGTTTCCCGACTGGAAAGCGGGCAGTGA
Protein Sequence
MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ
10xHis — 10xHis affinity tag 5319..5349 Fwd
DNA Sequence
CATCATCATCATCATCATCATCATCATCAC
Protein Sequence
HHHHHHHHHH
AviTag(TM) — peptide tag that allows for enzymatic b… 5352..5397 Fwd
DNA Sequence
GGCCTGAACGACATCTTCGAGGCTCAGAAAATCGAATGGCACGAA
Protein Sequence
GLNDIFEAQKIEWHE
HRV 3C site — recognition and cleavage site for human… 5397..5421 Fwd
DNA Sequence
CTGGAAGTTCTGTTCCAGGGGCCC
Protein Sequence
LEVLFQGP
ubiquitin Homo sapiens UBC human ubiquitin 5421..5649 Fwd
DNA Sequence
ATGCAAATCTTTGTTAAAACCCTGACCGGTAAAACCATTACACTGGAAGTTGAACCGAGCGATACCATTGAAAATGTGAAAGCCAAAATCCAGGACAAAGAAGGTATTCCGCCTGATCAGCAGCGTCTGATTTTTGCAGGTAAACAGCTGGAAGATGGTCGTACCCTGAGCGATTATAACATTCAGAAAGAAAGCACCCTGCATCTGGTTCTGCGTCTGCGTGGTGGT
Protein Sequence
MQIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLIFAGKQLEDGRTLSDYNIQKESTLHLVLRLRGG
BirA birA E. coli biotin protein ligase 5687..6650 Fwd
DNA Sequence
ATGAAGGATAACACTGTTCCGCTGAAACTGATCGCCCTGCTGGCCAATGGCGAGTTTCATAGCGGTGAACAGCTGGGTGAAACTTTAGGCATGTCACGTGCTGCTATCAACAAGCATATCCAGACCCTGCGTGATTGGGGTGTTGACGTATTTACCGTGCCGGGTAAAGGCTATTCCCTGCCGGAACCGATTCAGCTGCTGAACGCAAAACAAATTCTGGGTCAGCTGGACGGCGGTTCCGTAGCTGTGCTGCCGGTTATCGACTCTACTAACCAGTACCTCCTGGATCGCATTGGTGAACTGAAATCCGGGGACGCGTGCATCGCGGAGTATCAGCAAGCGGGCCGTGGTCGTCGCGGTCGTAAATGGTTTTCGCCGTTTGGTGCAAACCTGTACCTCAGCATGTTTTGGCGTCTGGAACAGGGTCCGGCAGCTGCAATCGGTTTGTCCCTGGTAATCGGTATCGTAATGGCGGAGGTTCTGCGTAAACTGGGCGCAGACAAAGTTCGTGTGAAATGGCCGAACGACCTTTACCTGCAAGATCGCAAACTGGCTGGTATCCTGGTTGAGCTGACCGGTAAAACCGGGGATGCTGCTCAGATCGTTATCGGTGCAGGTATCAACATGGCCATGCGTCGTGTAGAGGAAAGCGTGGTTAACCAGGGTTGGATTACCCTGCAGGAGGCGGGCATTAACCTGGACCGCAATACTCTGGCGGCTATGCTGATCCGCGAACTGCGTGCGGCACTGGAGCTGTTCGAACAGGAAGGTCTGGCTCCGTATCTTAGCCGCTGGGAAAAGCTCGATAACTTCATCAACCGTCCGGTTAAACTGATTATTGGTGATAAAGAAATCTTCGGCATTTCGCGCGGCATCGACAAACAGGGAGCCCTGCTTCTGGAACAGGATGGTATCATCAAACCGTGGATGGGCGGCGAAATTTCTTTACGCAGCGCTGAAAAA
Protein Sequence
MKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRDWGVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSGDACIAEYQQAGRGRRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRKLGADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQGWITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIGDKEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK
promoter 4
Label Gene Product Position Strand
AmpR promoter bla — 103..208 Fwd
DNA Sequence
CGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGT
lacI promoter lacI — 4840..4918 Rev
DNA Sequence
GACACCATCGAATGGCGCAAAACCTTTCGCGGTATGGCATGATAGCGCCCGGAAGAGAGTCAATTCAGGGTGGTGAAT
T7 promoter — — 5226..5245 Fwd
DNA Sequence
TAATACGACTCACTATAGG
tet promoter — — 6944..6973 Rev
DNA Sequence
TTGACAGCTTATCATCGATAAGCTTTAAT
protein_bind 1
Label Gene Product Position Strand
lac operator — — 5245..5270 Fwd
DNA Sequence
GGAATTGTGAGCGGATAACAATTCC
rep_origin 1
Label Gene Product Position Strand
ori — — 1239..1828 Fwd
DNA Sequence
TTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTCCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAA
terminator 1
Label Gene Product Position Strand
T7 terminator — — 6722..6770 Fwd
DNA Sequence
CTAGCATAACCCCTTGGGGCCTCTAAACGGGTCTTGAGGGGTTTTTTG