pY71-PT7-RiboJ-sfGFP-MGapt

129119 Synthetic Bacterial Expression

Plasmid Information

Purpose
Expresses a fluorescent reporter for the quantification of cell-free protein expression.
Depositor
Matthew Lux
Article
Cole et al ACS Synth Biol. 2019 Sep 20;8(9):2080-20
Insert
Superfolder GFP with Malachite Green aptamer
Tags
His6 and RiboJ Insulator
Promoter
T7
Available Since
April 22, 2022
Availability
Available

Sequence Information

Length
2,588 bp
Topology
circular
Molecule Type
other DNA

Features

CDS 5
Label Gene Product Position Strand
6xHis 6xHis affinity tag 742..760 Fwd
DNA Sequence
CATCACCATCACCATCAT
Protein Sequence
HHHHHH
TEV site tobacco etch virus (TEV) protease recog… 760..781 Fwd
DNA Sequence
GAGAATTTATACTTCCAGAGT
Protein Sequence
ENLYFQS
GFP11 11th beta-strand of superfolder GFP 1420..1468 Fwd
DNA Sequence
CGGGACCACATGGTTCTGCACGAATATGTGAATGCGGCAGGTATTACG
Protein Sequence
RDHMVLHEYVNAAGIT
Strep-Tag II peptide that binds Strep-Tactin(R), an … 1468..1492 Fwd
DNA Sequence
TGGAGCCATCCGCAGTTCGAAAAA
Protein Sequence
WSHPQFEK
KanR aph(3')-Ia aminoglycoside phosphotransferase 1659..2475 Rev
DNA Sequence
ATGAGCCATATTCAACGGGAAACGTCTTGCTCGAAGCCGCGATTAAATTCCAACATGGATGCTGATTTATATGGGTATAAATGGGCTCGCGATAATGTCGGGCAATCAGGTGCGACAATCTATCGATTGTATGGGAAGCCCGATGCGCCAGAGTTGTTTCTGAAACATGGCAAAGGTAGCGTTGCCAATGATGTTACAGATGAGATGGTCAGACTAAACTGGCTGACGGAATTTATGCCTCTTCCGACCATCAAGCATTTTATCCGTACTCCTGATGATGCATGGTTACTCACCACTGCGATCCCCGGGAAAACAGCATTCCAGGTATTAGAAGAATATCCTGATTCAGGTGAAAATATTGTTGATGCGCTGGCAGTGTTCCTGCGCCGGTTGCATTCGATTCCTGTTTGTAATTGTCCTTTTAACAGCGATCGCGTATTTCGTCTCGCTCAGGCGCAATCACGAATGAATAACGGTTTGGTTGATGCGAGTGATTTTGATGACGAGCGTAATGGCTGGCCTGTTGAACAAGTCTGGAAAGAAATGCATAAGCTTTTGCCATTCTCACCGGATTCAGTCGTCACTCATGGTGATTTCTCACTTGATAACCTTATTTTTGACGAGGGGAAATTAATAGGTTGTATTGATGTTGGACGAGTCGGAATCGCAGACCGATACCAGGATCTTGCCATCCTATGGAACTGCCTCGGTGAGTTTTCTCCTTCATTACAGAAACGGCTTTTTCAAAAATATGGTATTGATAATCCTGATATGAATAAATTGCAGTTTCATTTGATGCTCGATGAGTTTTTCTAA
Protein Sequence
MSHIQRETSCSKPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF
promoter 1
Label Gene Product Position Strand
T7 promoter 619..638 Fwd
DNA Sequence
TAATACGACTCACTATAGG
rep_origin 1
Label Gene Product Position Strand
ori 8..597 Fwd
DNA Sequence
TTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTTCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAA