pREC1010-opEbu1

240702 Synthetic Bacterial Expression

Plasmid Information

Purpose
RSF1010 origin of replication plasmid containing Ebu1 recombitron with extended a1 a2 regions with a donor in the ncRNA targeting lacZ locus in Citrobacter freundii ATCC 8090 expressed by Pm promoter
Depositor
Seth Shipman
Article
González-Delgado et al bioRxiv 2025.06.16.660010
Insert
Ebu1 RT, Ebu1 ncRNA, CspRecT and EcSSB
Mutation
Extended a1 a2 regions
Available Since
Aug. 7, 2025
Availability
Available

Sequence Information

Length
10,125 bp
Topology
circular
Molecule Type
other DNA
View on AddGene.org

Features

CDS 5
Label Gene Product Position Strand
KanR aph(3')-Ia aminoglycoside phosphotransferase 1672..2488 Fwd
DNA Sequence
ATGAGCCATATTCAGCGTGAAACGAGCTGTAGCCGTCCGCGTCTGAACAGCAACATGGATGCGGATCTGTATGGCTATAAATGGGCGCGTGATAACGTGGGTCAGAGCGGCGCGACCATTTATCGTCTGTATGGCAAACCGGATGCGCCGGAACTGTTTCTGAAACATGGCAAAGGCAGCGTGGCGAACGATGTGACCGATGAAATGGTGCGTCTGAACTGGCTGACCGAATTTATGCCGCTGCCGACCATTAAACATTTTATTCGCACCCCGGATGATGCGTGGCTGCTGACCACCGCGATTCCGGGCAAAACCGCGTTTCAGGTGCTGGAAGAATATCCGGATAGCGGCGAAAACATTGTGGATGCGCTGGCCGTGTTTCTGCGTCGTCTGCATAGCATTCCGGTGTGCAACTGCCCGTTTAACAGCGATCGTGTGTTTCGTCTGGCCCAGGCGCAGAGCCGTATGAACAACGGCCTGGTGGATGCGAGCGATTTTGATGATGAACGTAACGGCTGGCCGGTGGAACAGGTGTGGAAAGAAATGCATAAACTGCTGCCGTTTAGCCCGGATAGCGTGGTGACCCACGGCGATTTTAGCCTGGATAACCTGATTTTCGATGAAGGCAAACTGATTGGCTGCATTGATGTGGGCCGTGTGGGCATTGCGGATCGTTATCAGGATCTGGCCATTCTGTGGAACTGCCTGGGCGAATTTAGCCCGAGCCTGCAAAAACGTCTGTTTCAGAAATATGGCATTGATAATCCGGATATGAACAAACTGCAATTTCATCTGATGCTGGATGAATTTTTCTAA
Protein Sequence
MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF
RSF1010 RepC repC replication protein C of the broad-host… 2773..3625 Rev
DNA Sequence
GTGGTGAAGCCTAAGAACAAGCACAGCCTCAGCCACGTCCGGCACGACCCGGCGCACTGTCTGGCCCCCGGCCTGTTCCGTGCCCTCAAGCGGGGCGAGCGCAAGCGCAGCAAGCTGGACGTGACGTATGACTACGGCGACGGCAAGCGGATCGAGTTCAGCGGCCCGGAGCCGCTGGGCGCTGATGATCTGCGCATCCTGCAAGGGCTGGTGGCCATGGCTGGGCCTAATGGCCTAGTGCTTGGCCCGGAACCCAAGACCGAAGGCGGACGGCAGCTCCGGCTGTTCCTGGAACCCAAGTGGGAGGCCGTCACCGCTGATGCCATGGTGGTCAAAGGTAGCTATCGGGCGCTGGCAAAGGAAATCGGGGCAGAGGTCGATAGTGGTGGGGCGCTCAAGCACATACAGGACTGCATCGAGCGCCTTTGGAAGGTATCCATCATCGCCCAGAATGGCCGCAAGCGGCAGGGGTTTCGGCTGCTGTCGGAGTACGCCAGCGACGAGGCGGACGGGCGCCTGTACGTGGCCCTGAACCCCTTGATCGCGCAGGCCGTCATGGGTGGCGGCCAGCATGTGCGCATCAGCATGGACGAGGTGCGGGCGCTGGACAGCGAAACCGCCCGCCTGCTGCACCAGCGGCTGTGTGGCTGGATCGACCCCGGCAAAACCGGCAAGGCTTCCATAGATACCTTGTGCGGCTATGTCTGGCCGTCAGAGGCCAGTGGTTCGACCATGCGCAAGCGCCGCCAGCGGGTGCGCGAGGCGTTGCCGGAGCTGGTCGCGCTGGGCTGGACGGTAACCGAGTTCGCGGCGGGCAAGTACGACATCACCCGGCCCAAGGCGGCAGGCTGA
Protein Sequence
MVKPKNKHSLSHVRHDPAHCLAPGLFRALKRGERKRSKLDVTYDYGDGKRIEFSGPEPLGADDLRILQGLVAMAGPNGLVLGPEPKTEGGRQLRLFLEPKWEAVTADAMVVKGSYRALAKEIGAEVDSGGALKHIQDCIERLWKVSIIAQNGRKRQGFRLLSEYASDEADGRLYVALNPLIAQAVMGGGQHVRISMDEVRALDSETARLLHQRLCGWIDPGKTGKASIDTLCGYVWPSEASGSTMRKRRQRVREALPELVALGWTVTEFAAGKYDITRPKAAG
RSF1010 RepA repA replication protein A of the broad-host… 3611..4451 Rev
DNA Sequence
ATGGCTACCCATAAGCCTATCAATATTCTGGAGGCGTTCGCAGCAGCGCCGCCACCGCTGGACTACGTTTTGCCCAACATGGTGGCCGGTACGGTCGGGGCGCTGGTGTCGCCCGGTGGTGCCGGTAAATCCATGCTGGCCCTGCAACTGGCCGCACAGATTGCAGGCGGGCCGGATCTGCTGGAGGTGGGCGAACTGCCCACCGGCCCGGTGATCTACCTGCCCGCCGAAGACCCGCCCACCGCCATTCATCACCGCCTGCACGCCCTTGGGGCGCACCTCAGCGCCGAGGAACGGCAAGCCGTGGCTGACGGCCTGCTGATCCAGCCGCTGATCGGCAGCCTGCCCAACATCATGGCCCCGGAGTGGTTCGACGGCCTCAAGCGCGCCGCCGAGGGCCGCCGCCTGATGGTGCTGGACACGCTGCGCCGGTTCCACATCGAGGAAGAAAACGCCAGCGGCCCCATGGCCCAGGTCATCGGTCGCATGGAGGCCATCGCCGCCGATACCGGGTGCTCTATCGTGTTCCTGCACCATGCCAGCAAGGGCGCGGCCATGATGGGCGCAGGCGACCAGCAGCAGGCCAGCCGGGGCAGCTCGGTACTGGTCGATAACATCCGCTGGCAGTCCTACCTGTCGAGCATGACCAGCGCCGAGGCCGAGGAATGGGGTGTGGACGACGACCAGCGCCGGTTCTTCGTCCGCTTCGGTGTGAGCAAGGCCAACTATGGCGCACCGTTCGCTGATCGGTGGTTCAGGCGGCATGACGGCGGGGTGCTCAAGCCCGCCGTGCTGGAGAGGCAGCGCAAGAGCAAGGGGGTGCCCCGTGGTGAAGCCTAA
Protein Sequence
MATHKPINILEAFAAAPPPLDYVLPNMVAGTVGALVSPGGAGKSMLALQLAAQIAGGPDLLEVGELPTGPVIYLPAEDPPTAIHHRLHALGAHLSAEERQAVADGLLIQPLIGSLPNIMAPEWFDGLKRAAEGRRLMVLDTLRRFHIEEENASGPMAQVIGRMEAIAADTGCSIVFLHHASKGAAMMGAGDQQQASRGSSVLVDNIRWQSYLSSMTSAEAEEWGVDDDQRRFFVRFGVSKANYGAPFADRWFRRHDGGVLKPAVLERQRKSKGVPRGEA
RSF1010 RepB repB replication protein B of the broad-host… 4961..5933 Rev
DNA Sequence
ATGAAGAACGACAGGACTTTGCAGGCCATAGGCCGACAGCTCAAGGCCATGGGCTGTGAGCGCTTCGATATCGGCGTCAGGGACGCCACCACCGGCCAGATGATGAACCGGGAATGGTCAGCCGCCGAAGTGCTCCAGAACACGCCATGGCTCAAGCGGATGAATGCCCAGGGCAATGACGTGTATATCAGGCCCGCCGAGCAGGAGCGGCATGGTCTGGTGCTGGTGGACGACCTCAGCGAGTTTGACCTGGATGACATGAAAGCCGAGGGCCGGGAGCCTGCCCTGGTAGTGGAAACCAGCCCGAAGAACTATCAGGCATGGGTCAAGGTGGCCGACGCCGCAGGCGGTGAACTTCGGGGGCAGATTGCCCGGACGCTGGCCAGCGAGTACGACGCCGACCCGGCCAGCGCCGACAGCCGCCACTATGGCCGCTTGGCGGGCTTCACCAACCGCAAGGACAAGCACACCACCCGCGCCGGTTATCAGCCGTGGGTGCTGCTGCGTGAATCCAAGGGCAAGACCGCCACCGCTGGCCCGGCGCTGGTGCAGCAGGCTGGCCAGCAGATCGAGCAGGCCCAGCGGCAGCAGGAGAAGGCCCGCAGGCTGGCCAGCCTCGAACTGCCCGAGCGGCAGCTTAGCCGCCACCGGCGCACGGCGCTGGACGAGTACCGCAGCGAGATGGCCGGGCTGGTCAAGCGCTTCGGTGATGACCTCAGCAAGTGCGACTTTATCGCCGCGCAGAAGCTGGCCAGCCGGGGCCGCAGTGCCGAGGAAATCGGCAAGGCCATGGCCGAGGCCAGCCCAGCGCTGGCAGAGCGCAAGCCCGGCCACGAAGCGGATTACATCGAGCGCACCGTCAGCAAGGTCATGGGTCTGCCCAGCGTCCAGCTTGCGCGGGCCTAGCTGGCACGGGCACCGGCACCCCGCCAGCGAGGCATGGACAGGGGCGGGCCAGATTTCAGCATGTAG
Protein Sequence
MKNDRTLQAIGRQLKAMGCERFDIGVRDATTGQMMNREWSAAEVLQNTPWLKRMNAQGNDVYIRPAEQERHGLVLVDDLSEFDLDDMKAEGREPALVVETSPKNYQAWVKVADAAGGELRGQIARTLASEYDADPASADSRHYGRLAGFTNRKDKHTTRAGYQPWVLLRESKGKTATAGPALVQQAGQQIEQAQRQQEKARRLASLELPERQLSRHRRTALDEYRSEMAGLVKRFGDDLSKCDFIAAQKLASRGRSAEEIGKAMAEASPALAERKPGHEADYIERTVSKVMGLPSVQLARA*LARAPAPRQRGMDRGGPDFSM
XylS xylS XylS regulator encoded by the Pseudomon… 6582..7548 Rev
DNA Sequence
ATGGATTTTTGCTTATTGAACGAGAAAAGTCAGATCTTCGTCCACGCCGAGCCCTATGCAGTCTCCGATTATGTTAACCAGTATGTCGGTACGCACTCTATTCGCCTGCCCAAGGGCGGGCGCCCGGCAGGCAGGCTGCACCACAGAATCTTCGGATGCCTCGATCTGTGTCGAATCAGCTACGGCGGTAGCGTGAGGGTAATCTCGCCTGGATTAGAGACCTGTTATCATCTGCAAATAATACTCAAAGGCCATTGCCTGTGGCGTGGCCATGGCCAGGAGCACTATTTTGCGCCGGGCGAACTATTGCTGCTCAATCCGGATGACCAAGCCGACCTGACCTATTCAGAAGATTGCGAGAAATTTATCGTTAAATTGCCCTCAGTGGTCCTTGATCGGGCGTGCAGTGACAACAATTGGCACAAGCCGAGGGAGGGTATCCGTTTCGCCGCGCGACACAATCTCCAGCAACTCGATGGCTTTATCAATCTACTCGGGTTAGTTTGTGACGAAGCGGAACATACAAAGTCGATGCCTCGGGTCCAAGAGCACTATGCGGGGATCATCGCTTCCAAGCTGCTCGAAATGCTGGGCAGCAATGTCAGCCGTGAAATTTTCAGCAAAGGTAACCCGTCTTTCGAGCGAGTCGTTCAATTCATTGAGGAGAATCTCAAACGGAATATCAGCCTTGAGCGGTTAGCGGAGCTGGCGATGATGAGTCCACGCTCGCTCTACAATTTGTTCGAGAAGCACGCCGGCACCACGCCGAAGAACTACATCCGCAACCGCAAGCTCGAAAGCATCCGCGCCTGCTTGAACGATCCCAGTGCCAATGTGCGTAGTATAACTGAGATAGCCCTAGACTACGGCTTCTTACATTTGGGACGCTTCGCTGAAAACTATAGGAGCGCGTTCGGCGAGTTGCCTTCCGACACCCTGCGTCAATGCAAAAAGGAAGTGGCTTGA
Protein Sequence
MDFCLLNEKSQIFVHAEPYAVSDYVNQYVGTHSIRLPKGGRPAGRLHHRIFGCLDLCRISYGGSVRVISPGLETCYHLQIILKGHCLWRGHGQEHYFAPGELLLLNPDDQADLTYSEDCEKFIVKLPSVVLDRACSDNNWHKPREGIRFAARHNLQQLDGFINLLGLVCDEAEHTKSMPRVQEHYAGIIASKLLEMLGSNVSREIFSKGNPSFERVVQFIEENLKRNISLERLAELAMMSPRSLYNLFEKHAGTTPKNYIRNRKLESIRACLNDPSANVRSITEIALDYGFLHLGRFAENYRSAFGELPSDTLRQCKKEVA
RBS 1
Label Gene Product Position Strand
Shine-Dalgarno sequence — — 0..10125 Fwd
DNA Sequence
TAAGGAGGT
promoter 1
Label Gene Product Position Strand
Pm promoter — — 8447..8528 Fwd
DNA Sequence
AGTCCAGCCTTGCAAGAAGCGGATACAGGAGTGCAAAAAATGGCTATCTCTAGTAAGGCCTACCCCTTAGGCTTTATGCAA
rep_origin 1
Label Gene Product Position Strand
RSF1010 oriV — — 6039..6434 Fwd
DNA Sequence
GGCTGCACACGCGCCCCCACCCTTCGGGTAGGGGGAAAGGCCGCTAAAGCGGCTAAAAGCGCTCCAGCGTATTTCTGCGGGGTTTGGTGTGGGGTTTAGCGGGCTTTGCCCGCCTTTCCCCCTGCCGCGCAGCGGTGGGGCGGTGTGTAGCCTAGCGCAGCGAATAGACCAGCTATCCGGCCTCTGGCCGGGCATATTGGGCAAGGGCAGCAGCGCCCCACAAGGGCGCTGATAACCGCGCCTAGTGGATTATTCTTAGATAATCATGGATGGATTTTTCCAACACCCCGCCAGCCCCCGCCCCTGCTGGGTTTGCAGGTTTGGGGGCGTGACAGTTATTGCAGGGGTTCGTGACAGTTATTGCAGGGGGGCGTGACAGTTATTGCAGGGGTTCG
terminator 2
Label Gene Product Position Strand
lambda t0 terminator — — 1446..1541 Fwd
DNA Sequence
GACTCCTGTTGATAGATCCAGTAATGACCTCAGAACTCCATCTGGATTTGTTCAGAACGCTCGGTTGCCGCCGGGCGTTTTTTATTGGTGAGAAT
rrnB T1 terminator Escherichia coli rrnB — 6471..6558 Rev
DNA Sequence
CAAATAAAACGAAAGGCTCAGTCGAAAGACTGGGCCTTTCGTTTTATCTGTTGTTTGTCGGTGAACGCTCTCCTGAGTAGGACAAAT