pSCBE3-NG-HF1-Hypa

218162 Synthetic Bacterial Expression

Plasmid Information

Purpose
This plasmid harbors the base editor SCBE3-NG-HF1-Hypa along with an sgRNA cloning cassette, facilitating high-fidelity cytosine base editing at targets bearing NG PAM in Streptomyces.
Depositor
Yuhui Sun
Article
Wang et al Nat Commun. 2024 Jul 7;15(1):5687. doi:
Mutation
Mutations in the portion of SpCas9 : D10A / N497A…
Promoter
rpsL promoter
Available Since
May 13, 2024
Availability
Available

Sequence Information

Length
13,665 bp
Topology
circular
Molecule Type
other DNA
View on AddGene.org

Features

CDS 5
Label Gene Product Position Strand
ApmR aac(3)IV aminoglycoside 3-N-acetyltransferase ty… 3..804 Rev
DNA Sequence
ATGTCATCAGCGGTGGAGTGCAATGTCGTGCAATACGAATGGCGAAAAGCCGAGCTCATCGGTCAGCTTCTCAACCTTGGGGTTACCCCCGGCGGTGTGCTGCTGGTCCACAGCTCCTTCCGTAGCGTCCGGCCCCTCGAAGATGGGCCACTTGGACTGATCGAGGCCCTGCGTGCTGCGCTGGGTCCGGGAGGGACGCTCGTCATGCCCTCGTGGTCAGGTCTGGACGACGAGCCGTTCGATCCTGCCACGTCGCCCGTTACACCGGACCTTGGAGTTGTCTCTGACACATTCTGGCGCCTGCCAAATGTAAAGCGCAGCGCCCATCCATTTGCCTTTGCGGCAGCGGGGCCACAGGCAGAGCAGATCATCTCTGATCCATTGCCCCTGCCACCTCACTCGCCTGCAAGCCCGGTCGCCCGTGTCCATGAACTCGATGGGCAGGTACTTCTCCTCGGCGTGGGACACGATGCCAACACGACGCTGCATCTTGCCGAGTTGATGGCAAAGGTTCCCTATGGGGTGCCGAGACACTGCACCATTCTTCAGGATGGCAAGTTGGTACGCGTCGATTATCTCGAGAATGACCACTGCTGTGAGCGCTTTGCCTTGGCGGACAGGTGGCTCAAGGAGAAGAGCCTTCAGAAGGAAGGTCCAGTCGGTCATGCCTTTGCTCGGTTGATCCGCTCCCGCGACATTGTGGCGACAGCCCTGGGTCAACTGGGCCGAGATCCGTTGATCTTCCTGCATCCGCCAGAGGCGGGATGCGAAGAATGCGATGCCGCTCGCCAGTCGATTGGC
Protein Sequence
MSSAVECNVVQYEWRKAELIGQLLNLGVTPGGVLLVHSSFRSVRPLEDGPLGLIEALRAALGPGGTLVMPSWSGLDDEPFDPATSPVTPDLGVVSDTFWRLPNVKRSAHPFAFAAAGPQAEQIISDPLPLPPHSPASPVARVHELDGQVLLLGVGHDANTTLHLAELMAKVPYGVPRHCTILQDGKLVRVDYLENDHCCERFALADRWLKEKSLQKEGPVGHAFARLIRSRDIVATALGQLGRDPLIFLHPPEAGCEECDAARQSIG
AmpR bla beta-lactamase 1235..2096 Fwd
DNA Sequence
ATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTGTTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGCAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAA
Protein Sequence
MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW
APOBEC-1 — cytidine deaminase (C to U editing enzy… 6893..7580 Fwd
DNA Sequence
ATGTCGTCCGAGACCGGCCCGGTGGCCGTGGACCCGACCCTGCGTCGCCGCATCGAGCCGCACGAGTTCGAGGTGTTCTTCGACCCGCGGGAGCTGCGCAAGGAGACCTGCCTGCTGTACGAGATCAACTGGGGCGGCCGGCACTCGATCTGGCGCCACACCTCGCAGAACACCAACAAGCACGTCGAGGTGAACTTCATCGAGAAGTTCACCACCGAGCGGTACTTCTGCCCGAACACCCGCTGCTCGATCACCTGGTTCCTGTCCTGGTCCCCGTGCGGCGAGTGCTCCCGGGCGATCACCGAGTTCCTGTCCCGCTACCCGCACGTCACCCTGTTCATCTACATCGCCCGGCTGTACCACCACGCCGACCCGCGGAACCGGCAGGGCCTGCGCGACCTGATCTCCTCCGGCGTGACCATCCAGATCATGACCGAGCAGGAGTCCGGCTACTGCTGGCGGAACTTCGTCAACTACTCGCCGTCCAACGAGGCCCACTGGCCGCGGTACCCGCACCTGTGGGTGCGCCTGTACGTCCTGGAGCTGTACTGCATCATCCTGGGCCTGCCGCCGTGCCTGAACATCCTGCGTCGCAAGCAGCCGCAGCTGACCTTCTTCACCATCGCCCTGCAGTCCTGCCACTACCAGCGCCTGCCGCCGCACATCCTGTGGGCGACCGGCCTGAAG
Protein Sequence
MSSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCLLYEINWGGRHSIWRHTSQNTNKHVEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGECSRAITEFLSRYPHVTLFIYIARLYHHADPRNRQGLRDLISSGVTIQIMTEQESGYCWRNFVNYSPSNEAHWPRYPHLWVRLYVLELYCIILGLPPCLNILRRKQPQLTFFTIALQSCHYQRLPPHILWATGLK
UGI — uracil-DNA glycosylase inhibitor from a… 11741..11993 Fwd
DNA Sequence
ACCAACCTGTCGGACATCATCGAGAAGGAGACCGGCAAGCAGCTGGTGATCCAGGAGTCCATCCTGATGCTGCCGGAGGAAGTGGAGGAAGTGATCGGCAACAAGCCGGAGTCGGACATCCTGGTGCACACCGCCTACGACGAGTCGACCGACGAGAACGTCATGCTGCTGACCTCGGACGCCCCGGAGTACAAGCCGTGGGCCCTGGTCATCCAGGACTCCAACGGCGAGAACAAGATCAAGATGCTGTGA
Protein Sequence
TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTAYDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML
traJ traJ oriT-recognizing protein 12497..12869 Fwd
DNA Sequence
ATGGCTGATGAAACCAAGCCAACCAGGAAGGGCAGCCCACCTATCAAGGTGTACTGCCTTCCAGACGAACGAAGAGCGATTGAGGAAAAGGCGGCGGCGGCCGGCATGAGCCTGTCGGCCTACCTGCTGGCCGTCGGCCAGGGCTACAAAATCACGGGCGTCGTGGACTATGAGCACGTCCGCGAGCTGGCCCGCATCAATGGCGACCTGGGCCGCCTGGGCGGCCTGCTGAAACTCTGGCTCACCGACGACCCGCGCACGGCGCGGTTCGGTGATGCCACGATCCTCGCCCTGCTGGCGAAGATCGAAGAGAAGCAGGACGAGCTTGGCAAGGTCATGATGGGCGTGGTCCGCCCGAGGGCAGAGCCATGA
Protein Sequence
MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAVGQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEEKQDELGKVMMGVVRPRAEP
polyA_signal 1
Label Gene Product Position Strand
SV40 poly(A) signal — — 3239..3374 Fwd
DNA Sequence
GATCCAGACATGATAAGATACATTGATGAGTTTGGACAAACCACAACTAGAATGCAGTGAAAAAAATGCTTTATTTGTGAAATTTGTGATGCTATTGCTTTATTTGTAACCATTATAAGCTGCAATAAACAAGTT
promoter 3
Label Gene Product Position Strand
T3 promoter — — 997..1016 Rev
DNA Sequence
AATTAACCCTCACTAAAGG
AmpR promoter bla — 1130..1235 Fwd
DNA Sequence
CGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGT
rpsL promoter — — 6591..6893 Fwd
DNA Sequence
GCCCTGCAGGCGGAAGTCAGGTAGACACGACTTCCGCTAGTCCTTGCAAGGTCTGCTGACGTGAGGCGGGGCGGTCGTTTTTGACCGCCCTGCCTTCGTCATGTAGGCTCGCTCGCTGTGCCTGGCGTGTCATCAGACGCCCAGGTCCCGGTGCCGTGAGGCCCGGGCCATCGAGCCGGTGGTACGTGGCTGCGGTCCCCTTGTGAGGGCTGCGCGCCGTGTGCTGTCCGGCGCGCACAGCCTTGAATCCACCCGCGGGGGCCGGCCGGTCTCCGTGAGCTCGAGTAGACGACGGAGACGTA
rep_origin 1
Label Gene Product Position Strand
ori — — 2266..2855 Fwd
DNA Sequence
TTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTCCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAA