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