pgRNA-non-target

129465 Synthetic Bacterial Expression

Plasmid Information

Purpose
Derived from pSCB2-sgRNA. 20 nucleotide sequence added as sgRNA binding region by inverse PCR. Used as a negative control.
Depositor
Silvia Cardona
Article
Hogan et al ACS Synth Biol. 2019 Sep 6. doi: 10.1021
Insert
Non-target/random gRNA
Promoter
BBa_J23119 (SpeI)
Available Since
Sept. 27, 2019
Availability
Available

Sequence Information

Length
5,601 bp
Topology
circular
Molecule Type
other DNA
View on AddGene.org

Features

CDS 2
Label Gene Product Position Strand
pBBR1 Rep — replication protein for the broad-host-… 1355..2018 Rev
DNA Sequence
ATGGCCACGCAGTCCAGAGAAATCGGCATTCAAGCCAAGAACAAGCCCGGTCACTGGGTGCAAACGGAACGCAAAGCGCATGAGGCGTGGGCCGGGCTTATTGCGAGGAAACCCACGGCGGCAATGCTGCTGCATCACCTCGTGGCGCAGATGGGCCACCAGAACGCCGTGGTGGTCAGCCAGAAGACACTTTCCAAGCTCATCGGACGTTCTTTGCGGACGGTCCAATACGCAGTCAAGGACTTGGTGGCCGAGCGCTGGATCTCCGTCGTGAAGCTCAACGGCCCCGGCACCGTGTCGGCCTACGTGGTCAATGACCGCGTGGCGTGGGGCCAGCCCCGCGACCAGTTGCGCCTGTCGGTGTTCAGTGCCGCCGTGGTGGTTGATCACGACGACCAGGACGAATCGCTGTTGGGGCATGGCGACCTGCGCCGCATCCCGACCCTGTATCCGGGCGAGCAGCAACTACCGACCGGCCCCGGCGAGGAGCCGCCCAGCCAGCCCGGCATTCCGGGCATGGAACCAGACCTGCCAGCCTTGACCGAAACGGAGGAATGGGAACGGCGCGGGCAGCAGCGCCTGCCGATGCCCGATGAGCCGTGTTTTCTGGACGATGGCGAGCCGTTGGAGCCGCCGACACGGGTCACGCTGCCGCGCCGGTAG
Protein Sequence
MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLHHLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVVNDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEEPPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR
TpR dhfrIIb from plasmid R388 E. coli plasmid-associated dihydrofolat… 5283..5520 Rev
DNA Sequence
ATGGGTCAAAGTAGCGATGAAGCCAACGCTCCCGTTGCAGGGCAGTTTGCGCTTCCCCTGAGTGCCACCTTTGGCTTAGGGGATCGCGTACGCAAGAAATCTGGTGCCGCTTGGCAGGGTCAAGTCGTCGGTTGGTATTGCACAAAACTCACTCCTGAAGGCTATGCGGTCGAGTCCGAATCCCACCCAGGCTCAGTGCAAATTTATCCTGTGGCTGCACTTGAACGTGTGGCCTAA
Protein Sequence
MGQSSDEANAPVAGQFALPLSATFGLGDRVRKKSGAAWQGQVVGWYCTKLTPEGYAVESESHPGSVQIYPVAALERVA
promoter 3
Label Gene Product Position Strand
Pc promoter intI1 (promoter lies within the coding sequence) — 244..273 Rev
DNA Sequence
TTGACATAAGCCTGTTCGGTTCGTAAACT
J23119(SpeI) promoter — — 338..373 Fwd
DNA Sequence
TTGACAGCTAGCTCAGTCCTAGGTATAATACTAGT
cat promoter — — 4182..4285 Fwd
DNA Sequence
TGATCGGCACGTAAGAGGTTCCAACTTTCACCATAATGAAATAAGATCACTACCGGGCGTATTTTTTGAGTTATCGAGATTTTCAGGAGCTAAGGAAGCTAAA
rep_origin 1
Label Gene Product Position Strand
pBBR1 oriV — — 2018..2790 Fwd
DNA Sequence
GCTTATCTCCATGCGGTAGGGGTGCCGCACGGTTGCGGCACCATGCGCAATCAGCTGCAACTTTTCGGCAGCGCGACAACAATTATGCGTTGCGTAAAAGTGGCAGTCAATTACAGATTTTCTTTAACCTACGCAATGAGCTATTGCGGGGGGTGCCGCAATGAGCTGTTGCGTACCCCCCTTTTTTAAGTTGTTGATTTTTAAGTCTTTCGCATTTCGCCCTATATCTAGTTCTTTGGTGCCCAAAGAAGGGCACCCCTGCGGGGTTCCCCCACGCCTTCGGCGCGGCTCCCCCTCCGGCAAAAAGTGGCCCCTCCGGGGCTTGTTGATCGACTGCGCGGCCTTCGGCCTTGCCCAAGGTGGCGCTGCCCCCTTGGAACCCCCGCACTCGCCGCCGTGAGGCTCGGGGGGCAGGCGGGCGGGCTTCGCCCTTCGACTGCCCCCACTCGCATAGGCTTGGGTCGTTCCAGGCGCGTCAAGGCCAAGCCGCTGCGCGGTCGCTGCGCGAGCCTTGACCCGCCTTCCACTTGGTGTCCAACCGGCAAGCGAAGCGCGCAGGCCGCAGGCCGGAGGCTTTTCCCCAGAGAAAATTAAAAAAATTGATGGGGCAAGGCCGCAGGCCGCGCAGTTGGAGCCGGTGGGTATGTGGTCGAAGGCTGGGTAGCCGGTGGGCAATCCCTGTGGTCAAGCTCGTGGGCAGGCGCAGCCTGTCCATCAGCTTGTCCAGCAGGGTTGTCCACGGGCCGAGCGAAGCGAGCCAGCCGGTGGCCGC
terminator 2
Label Gene Product Position Strand
rrnB T1 terminator Escherichia coli rrnB — 684..771 Fwd
DNA Sequence
CAAATAAAACGAAAGGCTCAGTCGAAAGACTGGGCCTTTCGTTTTATCTGTTGTTTGTCGGTGAACGCTCTCCTGAGTAGGACAAAT
rrnB T2 terminator — — 862..890 Fwd
DNA Sequence
AGAAGGCCATCCTGACGGATGGCCTTTT