pORTMAGE311B

120418 Synthetic Bacterial Expression

Plasmid Information

Purpose
Expresses Lambda Beta and a dominant negative MutL E32K allele, controlled by XylS-Pm expression system for high precision and efficient MAGE experiments at 37°C. RSF1010 Ori; Broad host-range; KanR
Depositor
Csaba Pál
Article
Szili et al bioRxiv (2018)
Mutation
E32K mutation conferring dominant mutator phenoty…
Promoter
Pm
Available Since
Feb. 19, 2019
Availability
Available

Sequence Information

Length
9,878 bp
Topology
circular
Molecule Type
other DNA
View on AddGene.org

Features

CDS 6
Label Gene Product Position Strand
KanR aph(3')-Ia aminoglycoside phosphotransferase 2163..2979 Fwd
DNA Sequence
ATGAGCCATATTCAGCGTGAAACGAGCTGTAGCCGTCCGCGTCTGAACAGCAACATGGATGCGGATCTGTATGGCTATAAATGGGCGCGTGATAACGTGGGTCAGAGCGGCGCGACCATTTATCGTCTGTATGGCAAACCGGATGCGCCGGAACTGTTTCTGAAACATGGCAAAGGCAGCGTGGCGAACGATGTGACCGATGAAATGGTGCGTCTGAACTGGCTGACCGAATTTATGCCGCTGCCGACCATTAAACATTTTATTCGCACCCCGGATGATGCGTGGCTGCTGACCACCGCGATTCCGGGCAAAACCGCGTTTCAGGTGCTGGAAGAATATCCGGATAGCGGCGAAAACATTGTGGATGCGCTGGCCGTGTTTCTGCGTCGTCTGCATAGCATTCCGGTGTGCAACTGCCCGTTTAACAGCGATCGTGTGTTTCGTCTGGCCCAGGCGCAGAGCCGTATGAACAACGGCCTGGTGGATGCGAGCGATTTTGATGATGAACGTAACGGCTGGCCGGTGGAACAGGTGTGGAAAGAAATGCATAAACTGCTGCCGTTTAGCCCGGATAGCGTGGTGACCCACGGCGATTTTAGCCTGGATAACCTGATTTTCGATGAAGGCAAACTGATTGGCTGCATTGATGTGGGCCGTGTGGGCATTGCGGATCGTTATCAGGATCTGGCCATTCTGTGGAACTGCCTGGGCGAATTTAGCCCGAGCCTGCAAAAACGTCTGTTTCAGAAATATGGCATTGATAATCCGGATATGAACAAACTGCAATTTCATCTGATGCTGGATGAATTTTTCTAA
Protein Sequence
MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF
RSF1010 RepC repC replication protein C of the broad-host… 3264..4116 Rev
DNA Sequence
GTGGTGAAGCCTAAGAACAAGCACAGCCTCAGCCACGTCCGGCACGACCCGGCGCACTGTCTGGCCCCCGGCCTGTTCCGTGCCCTCAAGCGGGGCGAGCGCAAGCGCAGCAAGCTGGACGTGACGTATGACTACGGCGACGGCAAGCGGATCGAGTTCAGCGGCCCGGAGCCGCTGGGCGCTGATGATCTGCGCATCCTGCAAGGGCTGGTGGCCATGGCTGGGCCTAATGGCCTAGTGCTTGGCCCGGAACCCAAGACCGAAGGCGGACGGCAGCTCCGGCTGTTCCTGGAACCCAAGTGGGAGGCCGTCACCGCTGATGCCATGGTGGTCAAAGGTAGCTATCGGGCGCTGGCAAAGGAAATCGGGGCAGAGGTCGATAGTGGTGGGGCGCTCAAGCACATACAGGACTGCATCGAGCGCCTTTGGAAGGTATCCATCATCGCCCAGAATGGCCGCAAGCGGCAGGGGTTTCGGCTGCTGTCGGAGTACGCCAGCGACGAGGCGGACGGGCGCCTGTACGTGGCCCTGAACCCCTTGATCGCGCAGGCCGTCATGGGTGGCGGCCAGCATGTGCGCATCAGCATGGACGAGGTGCGGGCGCTGGACAGCGAAACCGCCCGCCTGCTGCACCAGCGGCTGTGTGGCTGGATCGACCCCGGCAAAACCGGCAAGGCTTCCATAGATACCTTGTGCGGCTATGTCTGGCCGTCAGAGGCCAGTGGTTCGACCATGCGCAAGCGCCGCCAGCGGGTGCGCGAGGCGTTGCCGGAGCTGGTCGCGCTGGGCTGGACGGTAACCGAGTTCGCGGCGGGCAAGTACGACATCACCCGGCCCAAGGCGGCAGGCTGA
Protein Sequence
MVKPKNKHSLSHVRHDPAHCLAPGLFRALKRGERKRSKLDVTYDYGDGKRIEFSGPEPLGADDLRILQGLVAMAGPNGLVLGPEPKTEGGRQLRLFLEPKWEAVTADAMVVKGSYRALAKEIGAEVDSGGALKHIQDCIERLWKVSIIAQNGRKRQGFRLLSEYASDEADGRLYVALNPLIAQAVMGGGQHVRISMDEVRALDSETARLLHQRLCGWIDPGKTGKASIDTLCGYVWPSEASGSTMRKRRQRVREALPELVALGWTVTEFAAGKYDITRPKAAG
RSF1010 RepA repA replication protein A of the broad-host… 4102..4942 Rev
DNA Sequence
ATGGCTACCCATAAGCCTATCAATATTCTGGAGGCGTTCGCAGCAGCGCCGCCACCGCTGGACTACGTTTTGCCCAACATGGTGGCCGGTACGGTCGGGGCGCTGGTGTCGCCCGGTGGTGCCGGTAAATCCATGCTGGCCCTGCAACTGGCCGCACAGATTGCAGGCGGGCCGGATCTGCTGGAGGTGGGCGAACTGCCCACCGGCCCGGTGATCTACCTGCCCGCCGAAGACCCGCCCACCGCCATTCATCACCGCCTGCACGCCCTTGGGGCGCACCTCAGCGCCGAGGAACGGCAAGCCGTGGCTGACGGCCTGCTGATCCAGCCGCTGATCGGCAGCCTGCCCAACATCATGGCCCCGGAGTGGTTCGACGGCCTCAAGCGCGCCGCCGAGGGCCGCCGCCTGATGGTGCTGGACACGCTGCGCCGGTTCCACATCGAGGAAGAAAACGCCAGCGGCCCCATGGCCCAGGTCATCGGTCGCATGGAGGCCATCGCCGCCGATACCGGGTGCTCTATCGTGTTCCTGCACCATGCCAGCAAGGGCGCGGCCATGATGGGCGCAGGCGACCAGCAGCAGGCCAGCCGGGGCAGCTCGGTACTGGTCGATAACATCCGCTGGCAGTCCTACCTGTCGAGCATGACCAGCGCCGAGGCCGAGGAATGGGGTGTGGACGACGACCAGCGCCGGTTCTTCGTCCGCTTCGGTGTGAGCAAGGCCAACTATGGCGCACCGTTCGCTGATCGGTGGTTCAGGCGGCATGACGGCGGGGTGCTCAAGCCCGCCGTGCTGGAGAGGCAGCGCAAGAGCAAGGGGGTGCCCCGTGGTGAAGCCTAA
Protein Sequence
MATHKPINILEAFAAAPPPLDYVLPNMVAGTVGALVSPGGAGKSMLALQLAAQIAGGPDLLEVGELPTGPVIYLPAEDPPTAIHHRLHALGAHLSAEERQAVADGLLIQPLIGSLPNIMAPEWFDGLKRAAEGRRLMVLDTLRRFHIEEENASGPMAQVIGRMEAIAADTGCSIVFLHHASKGAAMMGAGDQQQASRGSSVLVDNIRWQSYLSSMTSAEAEEWGVDDDQRRFFVRFGVSKANYGAPFADRWFRRHDGGVLKPAVLERQRKSKGVPRGEA
RSF1010 RepB repB replication protein B of the broad-host… 5452..6424 Rev
DNA Sequence
ATGAAGAACGACAGGACTTTGCAGGCCATAGGCCGACAGCTCAAGGCCATGGGCTGTGAGCGCTTCGATATCGGCGTCAGGGACGCCACCACCGGCCAGATGATGAACCGGGAATGGTCAGCCGCCGAAGTGCTCCAGAACACGCCATGGCTCAAGCGGATGAATGCCCAGGGCAATGACGTGTATATCAGGCCCGCCGAGCAGGAGCGGCATGGTCTGGTGCTGGTGGACGACCTCAGCGAGTTTGACCTGGATGACATGAAAGCCGAGGGCCGGGAGCCTGCCCTGGTAGTGGAAACCAGCCCGAAGAACTATCAGGCATGGGTCAAGGTGGCCGACGCCGCAGGCGGTGAACTTCGGGGGCAGATTGCCCGGACGCTGGCCAGCGAGTACGACGCCGACCCGGCCAGCGCCGACAGCCGCCACTATGGCCGCTTGGCGGGCTTCACCAACCGCAAGGACAAGCACACCACCCGCGCCGGTTATCAGCCGTGGGTGCTGCTGCGTGAATCCAAGGGCAAGACCGCCACCGCTGGCCCGGCGCTGGTGCAGCAGGCTGGCCAGCAGATCGAGCAGGCCCAGCGGCAGCAGGAGAAGGCCCGCAGGCTGGCCAGCCTCGAACTGCCCGAGCGGCAGCTTAGCCGCCACCGGCGCACGGCGCTGGACGAGTACCGCAGCGAGATGGCCGGGCTGGTCAAGCGCTTCGGTGATGACCTCAGCAAGTGCGACTTTATCGCCGCGCAGAAGCTGGCCAGCCGGGGCCGCAGTGCCGAGGAAATCGGCAAGGCCATGGCCGAGGCCAGCCCAGCGCTGGCAGAGCGCAAGCCCGGCCACGAAGCGGATTACATCGAGCGCACCGTCAGCAAGGTCATGGGTCTGCCCAGCGTCCAGCTTGCGCGGGCCGAGCTGGCACGGGCACCGGCACCCCGCCAGCGAGGCATGGACAGGGGCGGGCCAGATTTCAGCATGTAG
Protein Sequence
MKNDRTLQAIGRQLKAMGCERFDIGVRDATTGQMMNREWSAAEVLQNTPWLKRMNAQGNDVYIRPAEQERHGLVLVDDLSEFDLDDMKAEGREPALVVETSPKNYQAWVKVADAAGGELRGQIARTLASEYDADPASADSRHYGRLAGFTNRKDKHTTRAGYQPWVLLRESKGKTATAGPALVQQAGQQIEQAQRQQEKARRLASLELPERQLSRHRRTALDEYRSEMAGLVKRFGDDLSKCDFIAAQKLASRGRSAEEIGKAMAEASPALAERKPGHEADYIERTVSKVMGLPSVQLARAELARAPAPRQRGMDRGGPDFSM
XylS xylS XylS regulator encoded by the Pseudomon… 7073..8039 Rev
DNA Sequence
ATGGATTTTTGCTTATTGAACGAGAAAAGTCAGATCTTCGTCCACGCCGAGCCCTATGCAGTCTCCGATTATGTTAACCAGTATGTCGGTACGCACTCTATTCGCCTGCCCAAGGGCGGGCGCCCGGCAGGCAGGCTGCACCACAGAATCTTCGGATGCCTCGATCTGTGTCGAATCAGCTACGGCGGTAGCGTGAGGGTAATCTCGCCTGGATTAGAGACCTGTTATCATCTGCAAATAATACTCAAAGGCCATTGCCTGTGGCGTGGCCATGGCCAGGAGCACTATTTTGCGCCGGGCGAACTATTGCTGCTCAATCCGGATGACCAAGCCGACCTGACCTATTCAGAAGATTGCGAGAAATTTATCGTTAAATTGCCCTCAGTGGTCCTTGATCGGGCGTGCAGTGACAACAATTGGCACAAGCCGAGGGAGGGTATCCGTTTCGCCGCGCGACACAATCTCCAGCAACTCGATGGCTTTATCAATCTACTCGGGTTAGTTTGTGACGAAGCGGAACATACAAAGTCGATGCCTCGGGTCCAAGAGCACTATGCGGGGATCATCGCTTCCAAGCTGCTCGAAATGCTGGGCAGCAATGTCAGCCGTGAAATTTTCAGCAAAGGTAACCCGTCTTTCGAGCGAGTCGTTCAATTCATTGAGGAGAATCTCAAACGGAATATCAGCCTTGAGCGGTTAGCGGAGCTGGCGATGATGAGTCCACGCTCGCTCTACAATTTGTTCGAGAAGCACGCCGGCACCACGCCGAAGAACTACATCCGCAACCGCAAGCTCGAAAGCATCCGCGCCTGCTTGAACGATCCCAGTGCCAATGTGCGTAGTATAACTGAGATAGCCCTAGACTACGGCTTCTTACATTTGGGACGCTTCGCTGAAAACTATAGGAGCGCGTTCGGCGAGTTGCCTTCCGACACCCTGCGTCAATGCAAAAAGGAAGTGGCTTGA
Protein Sequence
MDFCLLNEKSQIFVHAEPYAVSDYVNQYVGTHSIRLPKGGRPAGRLHHRIFGCLDLCRISYGGSVRVISPGLETCYHLQIILKGHCLWRGHGQEHYFAPGELLLLNPDDQADLTYSEDCEKFIVKLPSVVLDRACSDNNWHKPREGIRFAARHNLQQLDGFINLLGLVCDEAEHTKSMPRVQEHYAGIIASKLLEMLGSNVSREIFSKGNPSFERVVQFIEENLKRNISLERLAELAMMSPRSLYNLFEKHAGTTPKNYIRNRKLESIRACLNDPSANVRSITEIALDYGFLHLGRFAENYRSAFGELPSDTLRQCKKEVA
Beta bet single-stranded DNA binding recombinase… 9091..9877 Fwd
DNA Sequence
ATGAGTACTGCACTCGCAACGCTGGCTGGGAAGCTGGCTGAACGTGTCGGCATGGATTCTGTCGACCCACAGGAACTGATCACCACTCTTCGCCAGACGGCATTTAAAGGTGATGCCAGCGATGCGCAGTTCATCGCATTACTGATCGTTGCCAACCAGTACGGCCTTAATCCGTGGACGAAAGAAATTTACGCCTTTCCTGATAAGCAGAATGGCATCGTTCCGGTGGTGGGCGTTGATGGCTGGTCCCGCATCATCAATGAAAACCAGCAGTTTGATGGCATGGACTTTGAGCAGGACAATGAATCCTGTACATGCCGGATTTACCGCAAGGACCGTAATCATCCGATCTGCGTTACCGAATGGATGGATGAATGCCGCCGCGAACCATTCAAAACTCGCGAAGGCAGAGAAATCACGGGGCCGTGGCAGTCGCATCCCAAACGGATGTTACGTCATAAAGCCATGATTCAGTGTGCCCGTCTGGCCTTCGGATTTGCTGGTATCTATGACAAGGATGAAGCCGAGCGCATTGTCGAAAATACTGCATACACTGCAGAACGTCAGCCGGAACGCGACATCACTCCGGTTAACGATGAAACCATGCAGGAGATTAACACTCTGCTGATCGCCCTGGATAAAACATGGGATGACGACTTATTGCCGCTCTGTTCCCAGATATTTCGCCGCGACATTCGTGCATCGTCAGAACTGACACAGGCCGAAGCAGTAAAAGCTCTTGGATTCCTGAAACAGAAAGCCGCAGAGCAGAAGGTGGCAGCATGA
Protein Sequence
MSTALATLAGKLAERVGMDSVDPQELITTLRQTAFKGDASDAQFIALLIVANQYGLNPWTKEIYAFPDKQNGIVPVVGVDGWSRIINENQQFDGMDFEQDNESCTCRIYRKDRNHPICVTEWMDECRREPFKTREGREITGPWQSHPKRMLRHKAMIQCARLAFGFAGIYDKDEAERIVENTAYTAERQPERDITPVNDETMQEINTLLIALDKTWDDDLLPLCSQIFRRDIRASSELTQAEAVKALGFLKQKAAEQKVAA
promoter 1
Label Gene Product Position Strand
Pm promoter — — 8938..9019 Fwd
DNA Sequence
AGTCCAGCCTTGCAAGAAGCGGATACAGGAGTGCAAAAAATGGCTATCTCTAGTAAGGCCTACCCCTTAGGCTTTATGCAA
rep_origin 1
Label Gene Product Position Strand
RSF1010 oriV — — 6530..6925 Fwd
DNA Sequence
GGCTGCACACGCGCCCCCACCCTTCGGGTAGGGGGAAAGGCCGCTAAAGCGGCTAAAAGCGCTCCAGCGTATTTCTGCGGGGTTTGGTGTGGGGTTTAGCGGGCTTTGCCCGCCTTTCCCCCTGCCGCGCAGCGGTGGGGCGGTGTGTAGCCTAGCGCAGCGAATAGACCAGCTATCCGGCCTCTGGCCGGGCATATTGGGCAAGGGCAGCAGCGCCCCACAAGGGCGCTGATAACCGCGCCTAGTGGATTATTCTTAGATAATCATGGATGGATTTTTCCAACACCCCGCCAGCCCCCGCCCCTGCTGGGTTTGCAGGTTTGGGGGCGTGACAGTTATTGCAGGGGTTCGTGACAGTTATTGCAGGGGGGCGTGACAGTTATTGCAGGGGTTCG
terminator 2
Label Gene Product Position Strand
lambda t0 terminator — — 1937..2032 Fwd
DNA Sequence
GACTCCTGTTGATAGATCCAGTAATGACCTCAGAACTCCATCTGGATTTGTTCAGAACGCTCGGTTGCCGCCGGGCGTTTTTTATTGGTGAGAAT
rrnB T1 terminator Escherichia coli rrnB — 6962..7049 Rev
DNA Sequence
CAAATAAAACGAAAGGCTCAGTCGAAAGACTGGGCCTTTCGTTTTATCTGTTGTTTGTCGGTGAACGCTCTCCTGAGTAGGACAAAT