cBEST4

234660 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
P3 and tcp830
Available Since
May 21, 2025
Availability
Available

Sequence Information

Length
9,821 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… 333..1020 Fwd
DNA Sequence
ATGTCCTCCGAGACCGGCCCCGTGGCCGTGGACCCGACCCTGCGTCGCCGCATCGAGCCGCACGAGTTCGAGGTGTTCTTCGACCCGCGCGAGCTGCGCAAGGAGACCTGCCTGCTGTACGAGATCAACTGGGGCGGCCGGCACTCCATCTGGCGCCACACCTCGCAGAACACCAACAAGCACGTCGAGGTCAACTTCATCGAGAAGTTCACCACCGAGCGGTACTTCTGCCCGAACACCCGCTGCTCCATCACCTGGTTCCTGTCCTGGTCCCCCTGCGGCGAGTGCTCCCGGGCCATCACCGAGTTCCTGTCCCGCTACCCGCACGTCACCCTGTTCATCTACATCGCCCGGCTGTACCACCACGCCGACCCGCGCAACCGGCAGGGCCTGCGCGACCTGATCTCCTCCGGCGTCACCATCCAGATCATGACCGAGCAGGAGTCCGGCTACTGCTGGCGGAACTTCGTCAACTACTCCCCGTCCAACGAGGCCCACTGGCCCCGCTACCCGCACCTGTGGGTCCGCCTGTACGTCCTGGAGCTGTACTGCATCATCCTGGGCCTGCCGCCCTGCCTGAACATCCTGCGTCGCAAGCAGCCGCAGCTGACCTTCTTCACCATCGCCCTCCAGTCCTGCCACTACCAGCGCCTGCCGCCCCACATCCTGTGGGCCACCGGCCTGAAG
Protein Sequence
MSSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCLLYEINWGGRHSIWRHTSQNTNKHVEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGECSRAITEFLSRYPHVTLFIYIARLYHHADPRNRQGLRDLISSGVTIQIMTEQESGYCWRNFVNYSPSNEAHWPRYPHLWVRLYVLELYCIILGLPPCLNILRRKQPQLTFFTIALQSCHYQRLPPHILWATGLK
Cas9(D10A) — nickase mutant of the Cas9 endonuclease… 1068..5169 Fwd
DNA Sequence
GACAAGAAGTACTCCATCGGCCTCGCGATCGGCACCAACTCCGTGGGCTGGGCGGTCATCACCGACGAGTACAAGGTCCCCTCCAAGAAGTTCAAGGTCCTGGGCAACACCGACCGGCACTCGATCAAGAAGAACCTGATCGGCGCCCTGCTCTTCGACAGCGGCGAGACCGCCGAGGCGACCCGCCTGAAGCGGACCGCGCGTCGCCGCTACACCCGGCGCAAGAACCGCATCTGCTACCTGCAGGAAATCTTCTCCAACGAGATGGCCAAGGTGGACGACTCGTTCTTCCACCGCCTGGAGGAGAGCTTCCTGGTGGAGGAGGACAAGAAGCACGAGCGCCACCCGATCTTCGGCAACATCGTGGACGAGGTGGCCTACCACGAGAAGTACCCCACCATCTACCACCTCCGCAAGAAGCTGGTGGACTCGACCGACAAGGCGGACCTGCGGCTCATCTACCTGGCCCTCGCGCACATGATCAAGTTCCGCGGCCACTTCCTCATCGAGGGCGACCTGAACCCGGACAACTCCGACGTGGACAAGCTCTTCATCCAGCTGGTGCAGACCTACAACCAGCTGTTCGAGGAGAACCCCATCAACGCCAGCGGCGTGGACGCCAAGGCGATCCTCTCCGCGCGCCTGAGCAAGTCCCGGCGCCTGGAGAACCTCATCGCCCAGCTGCCGGGCGAGAAGAAGAACGGCCTCTTCGGCAACCTGATCGCGCTGTCGCTCGGCCTGACCCCCAACTTCAAGAGCAACTTCGACCTGGCCGAGGACGCGAAGCTCCAGCTGTCCAAGGACACCTACGACGACGACCTGGACAACCTGCTCGCCCAGATCGGCGACCAGTACGCGGACCTCTTCCTGGCCGCGAAGAACCTCTCGGACGCCATCCTGCTCAGCGACATCCTGCGGGTCAACACCGAGATCACCAAGGCCCCGCTGTCGGCGAGCATGATCAAGCGGTACGACGAGCACCACCAGGACCTGACCCTGCTCAAGGCCCTCGTGCGCCAGCAGCTGCCCGAGAAGTACAAGGAAATCTTCTTCGACCAGTCCAAGAACGGCTACGCCGGCTACATCGACGGCGGCGCGTCGCAGGAGGAGTTCTACAAGTTCATCAAGCCGATCCTGGAGAAGATGGACGGCACCGAGGAGCTGCTCGTCAAGCTGAACCGCGAGGACCTGCTCCGCAAGCAGCGGACCTTCGACAACGGCTCCATCCCGCACCAGATCCACCTGGGCGAGCTCCACGCCATCCTCCGGCGCCAGGAGGACTTCTACCCCTTCCTGAAGGACAACCGCGAGAAGATCGAGAAGATCCTGACCTTCCGCATCCCGTACTACGTCGGCCCCCTGGCCCGCGGCAACTCCCGGTTCGCGTGGATGACCCGGAAGTCGGAGGAGACCATCACCCCGTGGAACTTCGAGGAGGTCGTGGACAAGGGCGCGTCCGCGCAGTCGTTCATCGAGCGCATGACCAACTTCGACAAGAACCTCCCGAACGAGAAGGTCCTGCCCAAGCACTCCCTGCTCTACGAGTACTTCACCGTGTACAACGAGCTGACCAAGGTCAAGTACGTGACCGAGGGCATGCGGAAGCCGGCCTTCCTGTCGGGCGAGCAGAAGAAGGCGATCGTGGACCTGCTCTTCAAGACCAACCGCAAGGTCACCGTGAAGCAGCTGAAGGAGGACTACTTCAAGAAGATCGAGTGCTTCGACTCCGTCGAGATCAGCGGCGTGGAGGACCGCTTCAACGCCTCCCTGGGCACCTACCACGACCTGCTCAAGATCATCAAGGACAAGGACTTCCTCGACAACGAGGAGAACGAGGACATCCTGGAGGACATCGTCCTCACCCTGACCCTCTTCGAGGACCGCGAGATGATCGAGGAGCGGCTCAAGACCTACGCCCACCTGTTCGACGACAAGGTGATGAAGCAGCTGAAGCGTCGCCGCTACACCGGCTGGGGCCGCCTCTCCCGGAAGCTGATCAACGGCATCCGGGACAAGCAGAGCGGCAAGACCATCCTGGACTTCCTCAAGTCCGACGGCTTCGCCAACCGCAACTTCATGCAGCTCATCCACGACGACAGCCTGACCTTCAAGGAGGACATCCAGAAGGCCCAGGTCTCGGGCCAGGGCGACAGCCTCCACGAGCACATCGCCAACCTGGCGGGCTCCCCGGCGATCAAGAAGGGCATCCTCCAGACCGTCAAGGTCGTGGACGAGCTGGTCAAGGTGATGGGCCGCCACAAGCCCGAGAACATCGTGATCGAGATGGCCCGGGAGAACCAGACCACCCAGAAGGGCCAGAAGAACTCGCGCGAGCGGATGAAGCGGATCGAGGAGGGCATCAAGGAGCTCGGCAGCCAGATCCTGAAGGAGCACCCGGTCGAGAACACCCAGCTGCAGAACGAGAAGCTGTACCTCTACTACCTGCAGAACGGCCGCGACATGTACGTGGACCAGGAGCTCGACATCAACCGGCTGTCCGACTACGACGTGGACCACATCGTGCCGCAGTCCTTCCTGAAGGACGACTCGATCGACAACAAGGTCCTGACCCGCTCGGACAAGAACCGGGGCAAGTCCGACAACGTGCCCTCGGAGGAGGTCGTGAAGAAGATGAAGAACTACTGGCGCCAGCTGCTCAACGCCAAGCTCATCACCCAGCGCAAGTTCGACAACCTGACCAAGGCCGAGCGGGGCGGCCTGAGCGAGCTCGACAAGGCGGGCTTCATCAAGCGCCAGCTGGTCGAGACCCGGCAGATCACCAAGCACGTGGCCCAGATCCTGGACTCCCGGATGAACACCAAGTACGACGAGAACGACAAGCTGATCCGCGAGGTCAAGGTGATCACCCTCAAGAGCAAGCTGGTCTCCGACTTCCGCAAGGACTTCCAGTTCTACAAGGTCCGGGAGATCAACAACTACCACCACGCCCACGACGCGTACCTGAACGCCGTCGTGGGCACCGCGCTGATCAAGAAGTACCCGAAGCTGGAGTCCGAGTTCGTCTACGGCGACTACAAGGTCTACGACGTGCGCAAGATGATCGCCAAGAGCGAGCAGGAGATCGGCAAGGCCACCGCGAAGTACTTCTTCTACTCCAACATCATGAACTTCTTCAAGACCGAGATCACCCTGGCCAACGGCGAGATCCGCAAGCGGCCCCTGATCGAGACCAACGGCGAGACCGGCGAGATCGTCTGGGACAAGGGCCGCGACTTCGCCACCGTCCGGAAGGTGCTGTCGATGCCGCAGGTCAACATCGTGAAGAAAACCGAGGTGCAGACCGGCGGCTTCAGCAAGGAGTCCATCCTCCCCAAGCGCAACAGCGACAAGCTGATCGCCCGGAAGAAGGACTGGGACCCGAAGAAGTACGGCGGCTTCGACAGCCCCACCGTCGCCTACTCCGTGCTGGTCGTGGCGAAGGTCGAGAAGGGCAAGAGCAAGAAGCTGAAGTCCGTGAAGGAGCTGCTCGGCATCACCATCATGGAGCGCTCCTCGTTCGAGAAGAACCCGATCGACTTCCTGGAGGCCAAGGGCTACAAGGAGGTCAAGAAGGACCTCATCATCAAGCTGCCCAAGTACAGCCTGTTCGAGCTGGAGAACGGCCGCAAGCGGATGCTCGCCTCCGCGGGCGAGCTGCAGAAGGGCAACGAGCTGGCCCTCCCGTCGAAGTACGTCAACTTCCTGTACCTCGCGTCCCACTACGAGAAGCTGAAGGGCTCGCCCGAGGACAACGAGCAGAAGCAGCTCTTCGTGGAGCAGCACAAGCACTACCTGGACGAGATCATCGAGCAGATCAGCGAGTTCAGCAAGCGCGTCATCCTGGCCGACGCGAACCTCGACAAGGTGCTGTCCGCCTACAACAAGCACCGCGACAAGCCGATCCGGGAGCAGGCGGAGAACATCATCCACCTGTTCACCCTCACCAACCTGGGCGCCCCCGCCGCGTTCAAGTACTTCGACACCACCATCGACCGCAAGCGGTACACCTCCACCAAGGAGGTCCTCGACGCGACCCTGATCCACCAGAGCATCACCGGCCTGTACGAGACCCGCATCGACCTGTCCCAGCTCGGCGGCGAC
Protein Sequence
DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGD
UGI — uracil-DNA glycosylase inhibitor from a… 5181..5433 Fwd
DNA Sequence
ACCAACCTGTCCGACATCATCGAGAAGGAGACCGGCAAGCAGCTGGTCATCCAGGAGTCCATCCTGATGCTGCCGGAGGAAGTCGAGGAAGTCATCGGCAACAAGCCCGAGTCCGACATCCTGGTCCACACCGCCTACGACGAGTCCACCGACGAGAACGTCATGCTGCTGACCTCGGACGCCCCCGAGTACAAGCCCTGGGCCCTGGTCATCCAGGACTCCAACGGCGAGAACAAGATCAAGATGCTGTGA
Protein Sequence
TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTAYDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML
promoter 5
Label Gene Product Position Strand
T3 promoter — — 3..22 Fwd
DNA Sequence
AATTAACCCTCACTAAAGG
gapdh promoter — — 48..333 Fwd
DNA Sequence
GCTGCTCCTTCGGTCGGACGTGCGTCTACGGGCACCTTACCGCAGCCGTCGGCTGTGCGACACGGACGGATCGGGCGAACTGGCCGATGCTGGGAGAAGCGCGCTGCTGTACGGCGCGCACCGGGTGCGGAGCCCCTCGGCGAGCGGTGTGAAACTTCTGTGAATGGCCTGTTCGGTTGCTTTTTTTATACGGCTGCCAGATAAGGCTTGCAGCATCTGGGCGGCTACCGCTATGATCGGGGCGTTCCTGCAATTCTTAGTGCGAGTATCTGAAAGGGGATACGC
T7 promoter — — 6016..6035 Fwd
DNA Sequence
TAATACGACTCACTATAGG
T3 promoter — — 6082..6101 Rev
DNA Sequence
AATTAACCCTCACTAAAGG
lac promoter — — 6169..6200 Rev
DNA Sequence
TTTACACTTTATGCTTCCGGCTCGTATGTTG
protein_bind 3
Label Gene Product Position Strand
tet operator tetO — 5801..5820 Fwd
DNA Sequence
TCCCTATCAGTGATAGAGA
lac operator — — 6145..6162 Fwd
DNA Sequence
TTGTTATCCGCTCACAA
CAP binding site — — 6214..6236 Fwd
DNA Sequence
ATGAGTGAGCTAACTCACATTA
rep_origin 1
Label Gene Product Position Strand
ori — — 6450..7039 Rev
DNA Sequence
TTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTTCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGGTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAA
terminator 2
Label Gene Product Position Strand
fd terminator — — 5613..5662 Rev
DNA Sequence
ACCGATACAATTAAAGGCTCCTTTTGGAGCCTTTTTTTTTGGAGATTTT
lambda t0 terminator — — 6374..6409 Fwd
DNA Sequence
TTCAGAACGCTCGGTTGCCGCCGGGCGTTTTTTAT