VR124 pETv2 10His-pG-Tn5 (E54K, L372P)

198468 Synthetic Bacterial Expression

Plasmid Information

Purpose
E. coli protein expression plasmid encoding hyperactive Tn5 transposase fused to protein G, in T7 Express lysY/Iq protein expression strain.
Depositor
Viviana Risca
Article
Soroczynski et al bioRxiv 2024.07.11.602973
Insert
10His-pG-Tn5 (E54K, L372P)
Tags
10His-pG
Mutation
E54K, L372P
Promoter
Consensus φ10 T7 Promoter
Available Since
April 14, 2023
Availability
Available

Sequence Information

Length
6,986 bp
Topology
circular
Molecule Type
other DNA
View on AddGene.org

Features

CDS 6
Label Gene Product Position Strand
10xHis — 10xHis affinity tag 103..133 Fwd
DNA Sequence
CATCATCATCATCATCACCACCACCATCAT
Protein Sequence
HHHHHHHHHH
GB1 — B1 domain of Streptococcal protein G 166..328 Fwd
DNA Sequence
TATAAATTGATCCTGAACGGCAAAACTTTGAAGGGCGAAACAACAACAGAAGCTGTTGACGCTGCTACCGCTGAAAAAGTATTTAAACAGTATGCAAACGACAACGGTGTTGATGGAGAATGGACATACGATGACGCAACCAAAACCTTCACTGTCACAGAA
Protein Sequence
YKLILNGKTLKGETTTEAVDAATAEKVFKQYANDNGVDGEWTYDDATKTFTVTE
Tn5 transposase — transposase from the bacterial Tn5 tran… 415..1840 Fwd
DNA Sequence
ATTACCAGTGCACTGCATCGTGCGGCGGATTGGGCGAAAAGCGTGTTTTCTAGTGCTGCGCTGGGTGATCCGCGTCGTACCGCGCGTCTGGTGAATGTTGCGGCGCAACTGGCCAAATATAGCGGCAAAAGCATTACCATTAGCAGCGAAGGCAGCAAAGCCATGCAGGAAGGCGCGTATCGTTTTATTCGTAATCCGAACGTGAGCGCGGAAGCGATTCGTAAAGCGGGTGCCATGCAGACCGTGAAACTGGCCCAGGAATTTCCGGAACTGCTGGCAATTGAAGATACCACCTCTCTGAGCTATCGTCATCAGGTGGCGGAAGAACTGGGCAAACTGGGTAGCATTCAGGATAAAAGCCGTGGTTGGTGGGTGCATAGCGTGCTGCTGCTGGAAGCGACCACCTTTCGTACCGTGGGCCTGCTGCATCAAGAATGGTGGATGCGTCCGGATGATCCGGCGGATGCGGATGAAAAAGAAAGCGGCAAATGGCTGGCCGCTGCTGCAACTTCGCGTCTGCGCATGGGCAGCATGATGAGCAACGTGATTGCGGTGTGCGATCGTGAAGCGGATATTCATGCGTATCTGCAAGATAAACTGGCCCATAACGAACGTTTTGTGGTGCGTAGCAAACATCCGCGTAAAGATGTGGAAAGCGGCCTGTATCTGTATGATCACCTGAAAAACCAGCCGGAACTGGGCGGCTATCAGATTAGCATTCCGCAGAAAGGCGTGGTGGATAAACGTGGCAAACGTAAAAACCGTCCGGCGCGTAAAGCGAGCCTGAGCCTGCGTAGCGGCCGTATTACCCTGAAACAGGGCAACATTACCCTGAACGCGGTGCTGGCCGAAGAAATTAATCCGCCGAAAGGCGAAACCCCGCTGAAATGGCTGCTGCTGACCAGCGAGCCGGTGGAAAGTCTGGCCCAAGCGCTGCGTGTGATTGATATTTATACCCATCGTTGGCGCATTGAAGAATTTCACAAAGCGTGGAAAACGGGTGCGGGTGCGGAACGTCAGCGTATGGAAGAACCGGATAACCTGGAACGTATGGTGAGCATTCTGAGCTTTGTGGCGGTGCGTCTGCTGCAACTGCGTGAATCTTTTACTCCGCCGCAAGCACTGCGTGCGCAGGGCCTGCTGAAAGAAGCGGAACACGTTGAAAGCCAGAGCGCGGAAACCGTGCTGACCCCGGATGAATGCCAACTGCTGGGCTATCTGGATAAAGGCAAACGCAAACGCAAAGAAAAAGCGGGCAGCCTGCAATGGGCGTATATGGCGATTGCGCGTCTGGGCGGCTTTATGGATAGCAAACGTACCGGCATTGCGAGCTGGGGTGCGCTGTGGGAAGGTTGGGAAGCGCTGCAAAGCAAACTGGATGGCTTTCTGGCCGCGAAAGACCTGATGGCGCAGGGCATTAAAATC
Protein Sequence
ITSALHRAADWAKSVFSSAALGDPRRTARLVNVAAQLAKYSGKSITISSEGSKAMQEGAYRFIRNPNVSAEAIRKAGAMQTVKLAQEFPELLAIEDTTSLSYRHQVAEELGKLGSIQDKSRGWWVHSVLLLEATTFRTVGLLHQEWWMRPDDPADADEKESGKWLAAAATSRLRMGSMMSNVIAVCDREADIHAYLQDKLAHNERFVVRSKHPRKDVESGLYLYDHLKNQPELGGYQISIPQKGVVDKRGKRKNRPARKASLSLRSGRITLKQGNITLNAVLAEEINPPKGETPLKWLLLTSEPVESLAQALRVIDIYTHRWRIEEFHKAWKTGAGAERQRMEEPDNLERMVSILSFVAVRLLQLRESFTPPQALRAQGLLKEAEHVESQSAETVLTPDECQLLGYLDKGKRKRKEKAGSLQWAYMAIARLGGFMDSKRTGIASWGALWEGWEALQSKLDGFLAAKDLMAQGIKI
KanR aph(3')-Ia aminoglycoside phosphotransferase 2562..3378 Rev
DNA Sequence
ATGAGCCATATTCAACGGGAAACGTCTTGCTCTAGGCCGCGATTAAATTCCAACATGGATGCTGATTTATATGGGTATAAATGGGCTCGCGATAATGTCGGGCAATCAGGTGCGACAATCTATCGATTGTATGGGAAGCCCGATGCGCCAGAGTTGTTTCTGAAACATGGCAAAGGTAGCGTTGCCAATGATGTTACAGATGAGATGGTCAGACTAAACTGGCTGACGGAATTTATGCCTCTTCCGACCATCAAGCATTTTATCCGTACTCCTGATGATGCATGGTTACTCACCACTGCGATCCCCGGGAAAACAGCATTCCAGGTATTAGAAGAATATCCTGATTCAGGTGAAAATATTGTTGATGCGCTGGCAGTGTTCCTGCGCCGGTTGCATTCGATTCCTGTTTGTAATTGTCCTTTTAACAGCGATCGCGTATTTCGTCTCGCTCAGGCGCAATCACGAATGAATAACGGTTTGGTTGATGCGAGTGATTTTGATGACGAGCGTAATGGCTGGCCTGTTGAACAAGTCTGGAAAGAAATGCATAAACTTTTGCCATTCTCACCGGATTCAGTCGTCACTCATGGTGATTTCTCACTTGATAACCTTATTTTTGACGAGGGGAAATTAATAGGTTGTATTGATGTTGGACGAGTCGGAATCGCAGACCGATACCAGGATCTTGCCATCCTATGGAACTGCCTCGGTGAGTTTTCTCCTTCATTACAGAAACGGCTTTTTCAAAAATATGGTATTGATAATCCTGATATGAATAAATTGCAGTTTCATTTGATGCTCGATGAGTTTTTCTAA
Protein Sequence
MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF
rop rop Rop protein, which maintains plasmids a… 4517..4709 Rev
DNA Sequence
GTGACCAAACAGGAAAAAACCGCCCTTAACATGGCCCGCTTTATCAGAAGCCAGACATTAACGCTTCTGGAGAAACTCAACGAGCTGGACGCGGATGAACAGGCAGACATCTGTGAATCGCTTCACGACCACGCTGATGAGCTTTACCGCAGCTGCCTCGCGCGTTTCGGTGATGACGGTGAAAACCTCTGA
Protein Sequence
MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHADELYRSCLARFGDDGENL
lacI lacI lac repressor 5517..6600 Rev
DNA Sequence
GTGAAACCAGTAACGTTATACGATGTCGCAGAGTATGCCGGTGTCTCTTATCAGACCGTTTCCCGCGTGGTGAACCAGGCCAGCCACGTTTCTGCGAAAACGCGGGAAAAAGTGGAAGCGGCGATGGCGGAGCTGAATTACATTCCCAACCGCGTGGCACAACAACTGGCGGGCAAACAGTCGTTGCTGATTGGCGTTGCCACCTCCAGTCTGGCCCTGCACGCGCCGTCGCAAATTGTCGCGGCGATTAAATCTCGCGCCGATCAACTGGGTGCCAGCGTGGTGGTGTCGATGGTAGAACGAAGCGGCGTCGAAGCCTGTAAAGCGGCGGTGCACAATCTTCTCGCGCAACGCGTCAGTGGGCTGATCATTAACTATCCGCTGGATGACCAGGATGCCATTGCTGTGGAAGCTGCCTGCACTAATGTTCCGGCGTTATTTCTTGATGTCTCTGACCAGACACCCATCAACAGTATTATTTTCTCCCATGAAGACGGTACGCGACTGGGCGTGGAGCATCTGGTCGCATTGGGTCACCAGCAAATCGCGCTGTTAGCGGGCCCATTAAGTTCTGTCTCGGCGCGTCTGCGTCTGGCTGGCTGGCATAAATATCTCACTCGCAATCAAATTCAGCCGATAGCGGAACGGGAAGGCGACTGGAGTGCCATGTCCGGTTTTCAACAAACCATGCAAATGCTGAATGAGGGCATCGTTCCCACTGCGATGCTGGTTGCCAACGATCAGATGGCGCTGGGCGCAATGCGCGCCATTACCGAGTCCGGGCTGCGCGTTGGTGCGGATATCTCGGTAGTGGGATACGACGATACCGAAGACAGCTCATGTTATATCCCGCCGTTAACCACCATCAAACAGGATTTTCGCCTGCTGGGGCAAACCAGCGTGGACCGCTTGCTGCAACTCTCTCAGGGCCAGGCGGTGAAGGGCAATCAGCTGTTGCCCGTCTCACTGGTGAAAAGAAAAACCACCCTGGCGCCCAATACGCAAACCGCCTCTCCCCGCGCGTTGGCCGATTCATTAATGCAGCTGGCACGACAGGTTTCCCGACTGGAAAGCGGGCAGTGA
Protein Sequence
MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ
promoter 2
Label Gene Product Position Strand
T7 promoter — — 0..19 Fwd
DNA Sequence
TAATACGACTCACTATAGG
lacI promoter lacI — 6600..6678 Rev
DNA Sequence
GACACCATCGAATGGCGCAAAACCTTTCGCGGTATGGCATGATAGCGCCCGGAAGAGAGTCAATTCAGGGTGGTGAAT
protein_bind 1
Label Gene Product Position Strand
lac operator — — 23..48 Fwd
DNA Sequence
GGAATTGTGAGCGGATAACAATTCC
rep_origin 2
Label Gene Product Position Strand
f1 ori — — 2014..2470 Fwd
DNA Sequence
ACGCGCCCTGTAGCGGCGCATTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCGTGACCGCTACACTTGCCAGCGCCCTAGCGCCCGCTCCTTTCGCTTTCTTCCCTTCCTTTCTCGCCACGTTCGCCGGCTTTCCCCGTCAAGCTCTAAATCGGGGGCTCCCTTTAGGGTTCCGATTTAGTGCTTTACGGCACCTCGACCCCAAAAAACTTGATTAGGGTGATGGTTCACGTAGTGGGCCATCGCCCTGATAGACGGTTTTTCGCCCTTTGACGTTGGAGTCCACGTTCTTTAATAGTGGACTCTTGTTCCAAACTGGAACAACACTCAACCCTATCTCGGTCTATTCTTTTGATTTATAAGGGATTTTGCCGATTTCGGCCTATTGGTTAAAAAATGAGCTGATTTAACAAAAATTTAACGCGAATTTTAACAAAATATTAACGTTTACAATTT
ori — — 3499..4088 Fwd
DNA Sequence
TTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTCCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAA
terminator 1
Label Gene Product Position Strand
T7 terminator — — 1930..1978 Fwd
DNA Sequence
CTAGCATAACCCCTTGGGGCCTCTAAACGGGTCTTGAGGGGTTTTTTG