cBEST2

234658 Synthetic Bacterial Expression

Plasmid Information

Purpose
Expresses cytosine base editor - spCas9n (D10A) fused to APOBEC1 and UGI, Include Golden Gate compatible cassette for sgRNA insertion
Depositor
Mingzi M. Zhang
Article
Lin et al Appl Environ Microbiol. 2025 Mar 10:e019
Promoter
kasO*17tss and kasOp*
Available Since
Nov. 13, 2025
Availability
Available

Sequence Information

Length
9,642 bp
Topology
circular
Molecule Type
other DNA
View on AddGene.org

Features

CDS 3
Label Gene Product Position Strand
APOBEC-1 — cytidine deaminase (C to U editing enzy… 145..832 Fwd
DNA Sequence
ATGTCCTCCGAGACCGGCCCCGTGGCCGTGGACCCGACCCTGCGTCGCCGCATCGAGCCGCACGAGTTCGAGGTGTTCTTCGACCCGCGCGAGCTGCGCAAGGAGACCTGCCTGCTGTACGAGATCAACTGGGGCGGCCGGCACTCCATCTGGCGCCACACCTCGCAGAACACCAACAAGCACGTCGAGGTCAACTTCATCGAGAAGTTCACCACCGAGCGGTACTTCTGCCCGAACACCCGCTGCTCCATCACCTGGTTCCTGTCCTGGTCCCCCTGCGGCGAGTGCTCCCGGGCCATCACCGAGTTCCTGTCCCGCTACCCGCACGTCACCCTGTTCATCTACATCGCCCGGCTGTACCACCACGCCGACCCGCGCAACCGGCAGGGCCTGCGCGACCTGATCTCCTCCGGCGTCACCATCCAGATCATGACCGAGCAGGAGTCCGGCTACTGCTGGCGGAACTTCGTCAACTACTCCCCGTCCAACGAGGCCCACTGGCCCCGCTACCCGCACCTGTGGGTCCGCCTGTACGTCCTGGAGCTGTACTGCATCATCCTGGGCCTGCCGCCCTGCCTGAACATCCTGCGTCGCAAGCAGCCGCAGCTGACCTTCTTCACCATCGCCCTCCAGTCCTGCCACTACCAGCGCCTGCCGCCCCACATCCTGTGGGCCACCGGCCTGAAG
Protein Sequence
MSSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCLLYEINWGGRHSIWRHTSQNTNKHVEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGECSRAITEFLSRYPHVTLFIYIARLYHHADPRNRQGLRDLISSGVTIQIMTEQESGYCWRNFVNYSPSNEAHWPRYPHLWVRLYVLELYCIILGLPPCLNILRRKQPQLTFFTIALQSCHYQRLPPHILWATGLK
Cas9(D10A) — nickase mutant of the Cas9 endonuclease… 880..4981 Fwd
DNA Sequence
GACAAGAAGTACTCCATCGGCCTCGCGATCGGCACCAACTCCGTGGGCTGGGCGGTCATCACCGACGAGTACAAGGTCCCCTCCAAGAAGTTCAAGGTCCTGGGCAACACCGACCGGCACTCGATCAAGAAGAACCTGATCGGCGCCCTGCTCTTCGACAGCGGCGAGACCGCCGAGGCGACCCGCCTGAAGCGGACCGCGCGTCGCCGCTACACCCGGCGCAAGAACCGCATCTGCTACCTGCAGGAAATCTTCTCCAACGAGATGGCCAAGGTGGACGACTCGTTCTTCCACCGCCTGGAGGAGAGCTTCCTGGTGGAGGAGGACAAGAAGCACGAGCGCCACCCGATCTTCGGCAACATCGTGGACGAGGTGGCCTACCACGAGAAGTACCCCACCATCTACCACCTCCGCAAGAAGCTGGTGGACTCGACCGACAAGGCGGACCTGCGGCTCATCTACCTGGCCCTCGCGCACATGATCAAGTTCCGCGGCCACTTCCTCATCGAGGGCGACCTGAACCCGGACAACTCCGACGTGGACAAGCTCTTCATCCAGCTGGTGCAGACCTACAACCAGCTGTTCGAGGAGAACCCCATCAACGCCAGCGGCGTGGACGCCAAGGCGATCCTCTCCGCGCGCCTGAGCAAGTCCCGGCGCCTGGAGAACCTCATCGCCCAGCTGCCGGGCGAGAAGAAGAACGGCCTCTTCGGCAACCTGATCGCGCTGTCGCTCGGCCTGACCCCCAACTTCAAGAGCAACTTCGACCTGGCCGAGGACGCGAAGCTCCAGCTGTCCAAGGACACCTACGACGACGACCTGGACAACCTGCTCGCCCAGATCGGCGACCAGTACGCGGACCTCTTCCTGGCCGCGAAGAACCTCTCGGACGCCATCCTGCTCAGCGACATCCTGCGGGTCAACACCGAGATCACCAAGGCCCCGCTGTCGGCGAGCATGATCAAGCGGTACGACGAGCACCACCAGGACCTGACCCTGCTCAAGGCCCTCGTGCGCCAGCAGCTGCCCGAGAAGTACAAGGAAATCTTCTTCGACCAGTCCAAGAACGGCTACGCCGGCTACATCGACGGCGGCGCGTCGCAGGAGGAGTTCTACAAGTTCATCAAGCCGATCCTGGAGAAGATGGACGGCACCGAGGAGCTGCTCGTCAAGCTGAACCGCGAGGACCTGCTCCGCAAGCAGCGGACCTTCGACAACGGCTCCATCCCGCACCAGATCCACCTGGGCGAGCTCCACGCCATCCTCCGGCGCCAGGAGGACTTCTACCCCTTCCTGAAGGACAACCGCGAGAAGATCGAGAAGATCCTGACCTTCCGCATCCCGTACTACGTCGGCCCCCTGGCCCGCGGCAACTCCCGGTTCGCGTGGATGACCCGGAAGTCGGAGGAGACCATCACCCCGTGGAACTTCGAGGAGGTCGTGGACAAGGGCGCGTCCGCGCAGTCGTTCATCGAGCGCATGACCAACTTCGACAAGAACCTCCCGAACGAGAAGGTCCTGCCCAAGCACTCCCTGCTCTACGAGTACTTCACCGTGTACAACGAGCTGACCAAGGTCAAGTACGTGACCGAGGGCATGCGGAAGCCGGCCTTCCTGTCGGGCGAGCAGAAGAAGGCGATCGTGGACCTGCTCTTCAAGACCAACCGCAAGGTCACCGTGAAGCAGCTGAAGGAGGACTACTTCAAGAAGATCGAGTGCTTCGACTCCGTCGAGATCAGCGGCGTGGAGGACCGCTTCAACGCCTCCCTGGGCACCTACCACGACCTGCTCAAGATCATCAAGGACAAGGACTTCCTCGACAACGAGGAGAACGAGGACATCCTGGAGGACATCGTCCTCACCCTGACCCTCTTCGAGGACCGCGAGATGATCGAGGAGCGGCTCAAGACCTACGCCCACCTGTTCGACGACAAGGTGATGAAGCAGCTGAAGCGTCGCCGCTACACCGGCTGGGGCCGCCTCTCCCGGAAGCTGATCAACGGCATCCGGGACAAGCAGAGCGGCAAGACCATCCTGGACTTCCTCAAGTCCGACGGCTTCGCCAACCGCAACTTCATGCAGCTCATCCACGACGACAGCCTGACCTTCAAGGAGGACATCCAGAAGGCCCAGGTCTCGGGCCAGGGCGACAGCCTCCACGAGCACATCGCCAACCTGGCGGGCTCCCCGGCGATCAAGAAGGGCATCCTCCAGACCGTCAAGGTCGTGGACGAGCTGGTCAAGGTGATGGGCCGCCACAAGCCCGAGAACATCGTGATCGAGATGGCCCGGGAGAACCAGACCACCCAGAAGGGCCAGAAGAACTCGCGCGAGCGGATGAAGCGGATCGAGGAGGGCATCAAGGAGCTCGGCAGCCAGATCCTGAAGGAGCACCCGGTCGAGAACACCCAGCTGCAGAACGAGAAGCTGTACCTCTACTACCTGCAGAACGGCCGCGACATGTACGTGGACCAGGAGCTCGACATCAACCGGCTGTCCGACTACGACGTGGACCACATCGTGCCGCAGTCCTTCCTGAAGGACGACTCGATCGACAACAAGGTCCTGACCCGCTCGGACAAGAACCGGGGCAAGTCCGACAACGTGCCCTCGGAGGAGGTCGTGAAGAAGATGAAGAACTACTGGCGCCAGCTGCTCAACGCCAAGCTCATCACCCAGCGCAAGTTCGACAACCTGACCAAGGCCGAGCGGGGCGGCCTGAGCGAGCTCGACAAGGCGGGCTTCATCAAGCGCCAGCTGGTCGAGACCCGGCAGATCACCAAGCACGTGGCCCAGATCCTGGACTCCCGGATGAACACCAAGTACGACGAGAACGACAAGCTGATCCGCGAGGTCAAGGTGATCACCCTCAAGAGCAAGCTGGTCTCCGACTTCCGCAAGGACTTCCAGTTCTACAAGGTCCGGGAGATCAACAACTACCACCACGCCCACGACGCGTACCTGAACGCCGTCGTGGGCACCGCGCTGATCAAGAAGTACCCGAAGCTGGAGTCCGAGTTCGTCTACGGCGACTACAAGGTCTACGACGTGCGCAAGATGATCGCCAAGAGCGAGCAGGAGATCGGCAAGGCCACCGCGAAGTACTTCTTCTACTCCAACATCATGAACTTCTTCAAGACCGAGATCACCCTGGCCAACGGCGAGATCCGCAAGCGGCCCCTGATCGAGACCAACGGCGAGACCGGCGAGATCGTCTGGGACAAGGGCCGCGACTTCGCCACCGTCCGGAAGGTGCTGTCGATGCCGCAGGTCAACATCGTGAAGAAAACCGAGGTGCAGACCGGCGGCTTCAGCAAGGAGTCCATCCTCCCCAAGCGCAACAGCGACAAGCTGATCGCCCGGAAGAAGGACTGGGACCCGAAGAAGTACGGCGGCTTCGACAGCCCCACCGTCGCCTACTCCGTGCTGGTCGTGGCGAAGGTCGAGAAGGGCAAGAGCAAGAAGCTGAAGTCCGTGAAGGAGCTGCTCGGCATCACCATCATGGAGCGCTCCTCGTTCGAGAAGAACCCGATCGACTTCCTGGAGGCCAAGGGCTACAAGGAGGTCAAGAAGGACCTCATCATCAAGCTGCCCAAGTACAGCCTGTTCGAGCTGGAGAACGGCCGCAAGCGGATGCTCGCCTCCGCGGGCGAGCTGCAGAAGGGCAACGAGCTGGCCCTCCCGTCGAAGTACGTCAACTTCCTGTACCTCGCGTCCCACTACGAGAAGCTGAAGGGCTCGCCCGAGGACAACGAGCAGAAGCAGCTCTTCGTGGAGCAGCACAAGCACTACCTGGACGAGATCATCGAGCAGATCAGCGAGTTCAGCAAGCGCGTCATCCTGGCCGACGCGAACCTCGACAAGGTGCTGTCCGCCTACAACAAGCACCGCGACAAGCCGATCCGGGAGCAGGCGGAGAACATCATCCACCTGTTCACCCTCACCAACCTGGGCGCCCCCGCCGCGTTCAAGTACTTCGACACCACCATCGACCGCAAGCGGTACACCTCCACCAAGGAGGTCCTCGACGCGACCCTGATCCACCAGAGCATCACCGGCCTGTACGAGACCCGCATCGACCTGTCCCAGCTCGGCGGCGAC
Protein Sequence
DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGD
UGI — uracil-DNA glycosylase inhibitor from a… 4993..5245 Fwd
DNA Sequence
ACCAACCTGTCCGACATCATCGAGAAGGAGACCGGCAAGCAGCTGGTCATCCAGGAGTCCATCCTGATGCTGCCGGAGGAAGTCGAGGAAGTCATCGGCAACAAGCCCGAGTCCGACATCCTGGTCCACACCGCCTACGACGAGTCCACCGACGAGAACGTCATGCTGCTGACCTCGGACGCCCCCGAGTACAAGCCCTGGGCCCTGGTCATCCAGGACTCCAACGGCGAGAACAAGATCAAGATGCTGTGA
Protein Sequence
TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTAYDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML
promoter 4
Label Gene Product Position Strand
T3 promoter — — 3..22 Fwd
DNA Sequence
AATTAACCCTCACTAAAGG
T7 promoter — — 5837..5856 Fwd
DNA Sequence
TAATACGACTCACTATAGG
T3 promoter — — 5903..5922 Rev
DNA Sequence
AATTAACCCTCACTAAAGG
lac promoter — — 5990..6021 Rev
DNA Sequence
TTTACACTTTATGCTTCCGGCTCGTATGTTG
protein_bind 2
Label Gene Product Position Strand
lac operator — — 5966..5983 Fwd
DNA Sequence
TTGTTATCCGCTCACAA
CAP binding site — — 6035..6057 Fwd
DNA Sequence
ATGAGTGAGCTAACTCACATTA
rep_origin 1
Label Gene Product Position Strand
ori — — 6271..6860 Rev
DNA Sequence
TTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTTCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGCGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGGTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAA
terminator 2
Label Gene Product Position Strand
fd terminator — — 5425..5474 Rev
DNA Sequence
ACCGATACAATTAAAGGCTCCTTTTGGAGCCTTTTTTTTTGGAGATTTT
lambda t0 terminator — — 6195..6230 Fwd
DNA Sequence
TTCAGAACGCTCGGTTGCCGCCGGGCGTTTTTTAT