pABE8e-protein

161788 Synthetic Bacterial Expression

Plasmid Information

Purpose
Rhamnose-inducible expression of 8xHIS-ABE8e in E. coli
Depositor
David Liu
Article
Huang et al Nat Protoc. 2021 Feb;16(2):1089-1128. do
Insert
ABE8e base editor
Tags
8xHIS and BPNLS
Promoter
pRhaBAD
Available Since
June 22, 2021
Availability
Available

Sequence Information

Length
7,095 bp
Topology
circular
Molecule Type
other DNA
View on AddGene.org

Features

CDS 4
Label Gene Product Position Strand
8xHis — 8xHis affinity tag 347..371 Fwd
DNA Sequence
CACCATCACCATCATCATCATCAC
Protein Sequence
HHHHHHHH
SV40 NLS — nuclear localization signal of SV40 (si… 404..425 Fwd
DNA Sequence
CCGAAAAAAAAGCGTAAAGTT
Protein Sequence
PKKKRKV
Cas9(D10A) — nickase mutant of the Cas9 endonuclease… 1019..5120 Fwd
DNA Sequence
GACAAAAAGTATTCGATTGGCCTGGCGATCGGCACGAACAGCGTCGGTTGGGCTGTTATTACCGACGAGTATAAAGTGCCGAGCAAGAAGTTTAAAGTCTTGGGTAACACCGATCGCCATAGCATCAAAAAGAATCTGATTGGTGCCCTGCTGTTTGATAGCGGCGAAACGGCAGAAGCGACCCGTCTGAAACGTACCGCACGTCGTCGCTACACTCGTCGTAAAAACCGCATTTGCTATTTGCAAGAGATCTTCAGCAATGAAATGGCCAAAGTTGACGACAGCTTCTTTCACCGTCTGGAGGAGAGCTTTCTGGTTGAAGAGGATAAGAAACACGAGCGTCACCCGATCTTTGGTAATATCGTTGACGAGGTCGCGTATCACGAGAAGTACCCGACGATCTACCACTTGCGTAAGAAACTGGTGGACAGCACCGATAAAGCCGATCTGCGTCTGATTTACTTGGCGTTGGCGCACATGATCAAGTTCCGCGGTCATTTCCTGATCGAGGGTGACCTGAACCCAGATAATTCCGACGTCGACAAGCTGTTCATCCAGCTGGTTCAGACCTACAACCAATTGTTTGAAGAAAACCCGATTAATGCATCTGGTGTCGACGCGAAGGCGATTCTGAGCGCGCGTCTGAGCAAGTCTCGTCGTCTGGAGAATCTGATCGCGCAATTGCCGGGTGAGAAGAAAAATGGTTTGTTTGGCAACTTGATCGCGCTGAGCCTGGGTCTGACCCCGAACTTTAAGAGCAATTTCGACCTGGCAGAAGATGCGAAACTGCAGCTGAGCAAAGACACCTATGATGACGACCTGGATAATCTCCTGGCACAAATTGGTGACCAGTATGCGGACCTGTTCCTGGCTGCGAAAAATCTGTCGGACGCCATTCTGCTGTCGGACATTCTGCGCGTTAACACTGAGATCACGAAAGCACCGCTGTCTGCGAGCATGATTAAACGTTATGACGAGCACCATCAAGACCTGACCCTGCTGAAGGCCCTGGTCCGTCAGCAACTGCCGGAGAAGTATAAAGAAATTTTTTTCGATCAGAGCAAGAACGGTTACGCAGGCTATATCGATGGCGGTGCGAGCCAAGAAGAGTTTTACAAATTCATTAAACCGATTTTGGAGAAGATGGACGGTACCGAGGAGTTGCTGGTGAAACTGAATCGCGAAGATCTGCTGCGCAAACAACGTACGTTCGATAACGGTAGCATCCCACACCAGATTCACCTGGGTGAGCTGCACGCGATTCTGCGTCGTCAGGAAGATTTCTACCCATTCCTGAAAGATAATCGTGAAAAGATTGAGAAAATCTTGACGTTCCGCATTCCGTACTACGTCGGTCCGCTGGCACGTGGCAACAGCCGTTTCGCTTGGATGACGCGCAAGAGCGAGGAAACTATCACGCCGTGGAACTTTGAGGAAGTCGTCGACAAAGGTGCGAGCGCGCAAAGCTTTATTGAGCGCATGACCAACTTTGACAAGAATCTGCCGAACGAGAAAGTCTTGCCAAAGCACAGCCTGCTGTACGAGTATTTCACGGTGTACAATGAACTGACCAAAGTTAAGTATGTCACCGAGGGTATGCGTAAACCAGCGTTCCTGTCCGGTGAACAGAAGAAAGCAATTGTGGATCTGTTGTTTAAGACGAACCGTAAAGTCACGGTGAAGCAGTTGAAAGAGGATTACTTCAAAAAGATCGAATGCTTTGACAGCGTGGAAATTAGCGGTGTTGAGGACCGTTTCAACGCCAGCTTGGGCACCTACCATGATCTGCTGAAAATCATTAAGGACAAGGACTTCCTGGATAACGAAGAAAACGAGGATATTCTGGAGGATATTGTCCTGACGCTGACCCTCTTCGAAGATCGCGAAATGATTGAAGAGAGACTGAAAACTTACGCCCACTTGTTTGACGATAAAGTCATGAAACAACTGAAGCGCCGTCGTTACACCGGTTGGGGTCGTCTGAGCCGTAAGCTGATCAACGGTATCCGTGATAAGCAATCTGGCAAAACGATCCTGGACTTTCTGAAAAGCGATGGTTTCGCGAACCGCAACTTCATGCAGCTTATTCACGACGATTCTCTGACCTTTAAGGAGGACATCCAAAAAGCCCAGGTTAGCGGTCAGGGCGATTCCCTGCATGAGCATATTGCTAACTTAGCGGGTAGCCCGGCAATCAAAAAGGGCATTCTGCAAACCGTTAAGGTTGTGGACGAGTTGGTTAAGGTCATGGGCCGTCATAAGCCGGAGAATATCGTTATTGAGATGGCGCGCGAGAATCAGACCACGCAGAAAGGCCAAAAGAATAGCCGTGAACGTATGAAACGTATTGAAGAGGGTATCAAAGAGCTGGGCAGCCAAATCTTGAAGGAACATCCGGTGGAGAACACGCAGCTGCAGAATGAAAAGCTGTACCTGTACTATCTGCAAAACGGTCGTGATATGTACGTTGACCAAGAGCTGGACATCAACCGCCTGAGCGATTATGATGTTGATCACATTGTGCCGCAATCTTTTCTGAAGGACGACAGCATCGACAACAAAGTTTTGACCCGTAGCGACAAGAATCGCGGTAAAAGCGATAATGTTCCGAGCGAAGAGGTCGTTAAGAAGATGAAAAACTATTGGCGTCAGTTGCTGAATGCAAAACTGATTACCCAACGCAAATTCGACAATTTGACTAAAGCGGAGCGTGGTGGTCTGAGCGAGCTGGACAAGGCGGGTTTCATTAAGCGTCAACTGGTTGAAACTCGTCAGATCACGAAGCACGTGGCGCAGATTCTGGACAGCCGCATGAATACTAAGTACGATGAAAATGATAAACTCATTCGTGAGGTCAAGGTCATTACCCTGAAGTCTAAGCTCGTCAGCGATTTTCGTAAAGACTTCCAGTTCTATAAGGTGCGCGAAATCAACAATTACCACCACGCCCATGATGCATATCTGAACGCCGTGGTGGGTACGGCTCTGATCAAGAAATATCCGAAACTGGAGTCCGAGTTTGTCTATGGTGACTACAAAGTGTATGACGTCCGCAAAATGATCGCCAAATCGGAACAAGAGATTGGCAAGGCAACCGCGAAGTACTTTTTCTATTCCAACATTATGAACTTTTTCAAGACCGAGATCACCCTGGCAAACGGTGAGATCCGTAAACGTCCGCTGATCGAAACTAACGGTGAAACCGGTGAAATCGTGTGGGACAAAGGTCGTGACTTTGCAACCGTGCGTAAAGTGCTGTCGATGCCACAGGTCAACATCGTTAAGAAAACCGAAGTTCAAACTGGTGGCTTTAGCAAGGAGAGCATTCTGCCTAAGCGCAACAGCGACAAGCTGATTGCACGTAAGAAGGACTGGGACCCGAAGAAGTACGGCGGCTTTGATTCCCCGACGGTTGCGTATAGCGTTCTCGTTGTGGCGAAAGTCGAGAAAGGCAAGAGCAAAAAGCTGAAAAGCGTTAAAGAATTGCTGGGTATCACGATTATGGAGCGTAGCAGCTTCGAAAAGAATCCTATTGATTTCTTGGAAGCAAAAGGTTACAAGGAGGTGAAGAAAGACTTGATCATCAAACTGCCGAAGTATTCCCTGTTCGAGCTGGAAAACGGCCGCAAACGTATGCTGGCCAGCGCTGGTGAACTGCAAAAGGGCAACGAACTGGCTCTGCCGAGCAAATATGTGAATTTCCTGTACCTGGCCAGCCACTACGAGAAACTGAAGGGCTCCCCGGAAGATAACGAGCAGAAGCAGCTGTTCGTTGAGCAGCACAAGCATTACCTGGACGAGATTATCGAGCAGATCAGCGAGTTTAGCAAACGCGTGATTCTGGCGGATGCGAATTTGGATAAAGTTCTGTCGGCCTATAATAAACACCGCGACAAACCGATTCGCGAGCAGGCGGAGAACATTATCCACTTGTTCACCCTGACCAATCTGGGTGCTCCGGCTGCTTTCAAATACTTTGATACCACGATTGATCGCAAAAGATATACGTCCACCAAAGAGGTACTGGACGCGACCCTGATCCACCAGAGCATCACCGGCTTGTACGAAACGCGCATCGATCTGTCTCAACTGGGCGGTGAC
Protein Sequence
DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGD
SV40 NLS — nuclear localization signal of SV40 (si… 5165..5186 Fwd
DNA Sequence
CCGAAAAAGAAACGCAAGGTG
Protein Sequence
PKKKRKV
promoter 2
Label Gene Product Position Strand
cat promoter — — 0..91 Rev
DNA Sequence
TGATCGGCACGTAAGAGGTTCCAACTTTCACCATAATGAAATAAGATCACTACCGGGCGTATTTTTTGAGTTATCGAGATTTTCAGGAGCT
rhaB promoter — — 194..313 Fwd
DNA Sequence
CACCACAATTCAGCAAATTGTGAACATCATCACGTTCATCTTTCCCTGGTTGCCAATGGCCCATTTTCCTGTCAGTAACGAGAAGGTCGCGAATTCAGGCGCTTTTTAGACTGGTCGTA
rep_origin 1
Label Gene Product Position Strand
p15A ori — — 5424..5970 Fwd
DNA Sequence
TTGAGATCGTTTTGGTCTGCGCGTAATCTCTTGCTCTGAAAACGAAAAAACCGCCTTGCAGGGCGGTTTTTCGAAGGTTCTCTGAGCTACCAACTCTTTGAACCGAGGTAACTGGCTTGGAGGAGCGCAGTCACCAAAACTTGTCCTTTCAGTTTAGCCTTAACCGGCGCATGACTTCAAGACTAACTCCTCTAAATCAATTACCAGTGGCTGCTGCCAGTGGTGCTTTTGCATGTCTTTCCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGACTGAACGGGGGGTTCGTGCATACAGTCCAGCTTGGAGCGAACTGCCTACCCGGAACTGAGTGTCAGGCGTGGAATGAGACAAACGCGGCCATAACAGCGGAATGACACCGGTAAACCGAAAGGCAGGAACAGGAGAGCGCACGAGGGAGCCGCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCACTGATTTGAGCGTCAGATTTCGTGATGCTTGTCAGGGGGGCGGAGCCTATGGAAA
terminator 4
Label Gene Product Position Strand
tonB terminator — — 99..130 Rev
DNA Sequence
AGTCAAAAGCCTCCGACCGGAGGCTTTTGAC
T7 terminator — — 5220..5268 Fwd
DNA Sequence
CTAGCATAACCCCTTGGGGCCTCTAAACGGGTCTTGAGGGGTTTTTTG
T3Te terminator — — 5365..5395 Fwd
DNA Sequence
GGCTCACCTTCACGGGTGGGCCTTTCTTCG
T7Te terminator — — 6234..6262 Rev
DNA Sequence
GGCTCACCTTCGGGTGGGCCTTTCTGCG