pSCB2-sgRNA

129463 Synthetic Bacterial Expression

Plasmid Information

Purpose
Template plasmid for guide sequence insertion via inverse PCR. Derived from pSCrhaB2-gRNA (see paper).
Depositor
Silvia Cardona
Article
Hogan et al ACS Synth Biol. 2019 Sep 6. doi: 10.1021
Insert
pgRNA
Promoter
BBa_J23119 (SpeI)
Available Since
Sept. 30, 2019
Availability
Available

Sequence Information

Length
5,602 bp
Topology
circular
Molecule Type
other DNA

Features

CDS 2
Label Gene Product Position Strand
pBBR1 Rep replication protein for the broad-host-… 845..1508 Rev
DNA Sequence
ATGGCCACGCAGTCCAGAGAAATCGGCATTCAAGCCAAGAACAAGCCCGGTCACTGGGTGCAAACGGAACGCAAAGCGCATGAGGCGTGGGCCGGGCTTATTGCGAGGAAACCCACGGCGGCAATGCTGCTGCATCACCTCGTGGCGCAGATGGGCCACCAGAACGCCGTGGTGGTCAGCCAGAAGACACTTTCCAAGCTCATCGGACGTTCTTTGCGGACGGTCCAATACGCAGTCAAGGACTTGGTGGCCGAGCGCTGGATCTCCGTCGTGAAGCTCAACGGCCCCGGCACCGTGTCGGCCTACGTGGTCAATGACCGCGTGGCGTGGGGCCAGCCCCGCGACCAGTTGCGCCTGTCGGTGTTCAGTGCCGCCGTGGTGGTTGATCACGACGACCAGGACGAATCGCTGTTGGGGCATGGCGACCTGCGCCGCATCCCGACCCTGTATCCGGGCGAGCAGCAACTACCGACCGGCCCCGGCGAGGAGCCGCCCAGCCAGCCCGGCATTCCGGGCATGGAACCAGACCTGCCAGCCTTGACCGAAACGGAGGAATGGGAACGGCGCGGGCAGCAGCGCCTGCCGATGCCCGATGAGCCGTGTTTTCTGGACGATGGCGAGCCGTTGGAGCCGCCGACACGGGTCACGCTGCCGCGCCGGTAG
Protein Sequence
MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLHHLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVVNDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEEPPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR
TpR dhfrIIb from plasmid R388 E. coli plasmid-associated dihydrofolat… 4773..5010 Rev
DNA Sequence
ATGGGTCAAAGTAGCGATGAAGCCAACGCTCCCGTTGCAGGGCAGTTTGCGCTTCCCCTGAGTGCCACCTTTGGCTTAGGGGATCGCGTACGCAAGAAATCTGGTGCCGCTTGGCAGGGTCAAGTCGTCGGTTGGTATTGCACAAAACTCACTCCTGAAGGCTATGCGGTCGAGTCCGAATCCCACCCAGGCTCAGTGCAAATTTATCCTGTGGCTGCACTTGAACGTGTGGCCTAA
Protein Sequence
MGQSSDEANAPVAGQFALPLSATFGLGDRVRKKSGAAWQGQVVGWYCTKLTPEGYAVESESHPGSVQIYPVAALERVA
promoter 3
Label Gene Product Position Strand
cat promoter 3672..3775 Fwd
DNA Sequence
TGATCGGCACGTAAGAGGTTCCAACTTTCACCATAATGAAATAAGATCACTACCGGGCGTATTTTTTGAGTTATCGAGATTTTCAGGAGCTAAGGAAGCTAAA
Pc promoter intI1 (promoter lies within the coding sequence) 5335..5364 Rev
DNA Sequence
TTGACATAAGCCTGTTCGGTTCGTAAACT
J23119(SpeI) promoter 5429..5464 Fwd
DNA Sequence
TTGACAGCTAGCTCAGTCCTAGGTATAATACTAGT
rep_origin 1
Label Gene Product Position Strand
pBBR1 oriV 1508..2280 Fwd
DNA Sequence
GCTTATCTCCATGCGGTAGGGGTGCCGCACGGTTGCGGCACCATGCGCAATCAGCTGCAACTTTTCGGCAGCGCGACAACAATTATGCGTTGCGTAAAAGTGGCAGTCAATTACAGATTTTCTTTAACCTACGCAATGAGCTATTGCGGGGGGTGCCGCAATGAGCTGTTGCGTACCCCCCTTTTTTAAGTTGTTGATTTTTAAGTCTTTCGCATTTCGCCCTATATCTAGTTCTTTGGTGCCCAAAGAAGGGCACCCCTGCGGGGTTCCCCCACGCCTTCGGCGCGGCTCCCCCTCCGGCAAAAAGTGGCCCCTCCGGGGCTTGTTGATCGACTGCGCGGCCTTCGGCCTTGCCCAAGGTGGCGCTGCCCCCTTGGAACCCCCGCACTCGCCGCCGTGAGGCTCGGGGGGCAGGCGGGCGGGCTTCGCCCTTCGACTGCCCCCACTCGCATAGGCTTGGGTCGTTCCAGGCGCGTCAAGGCCAAGCCGCTGCGCGGTCGCTGCGCGAGCCTTGACCCGCCTTCCACTTGGTGTCCAACCGGCAAGCGAAGCGCGCAGGCCGCAGGCCGGAGGCTTTTCCCCAGAGAAAATTAAAAAAATTGATGGGGCAAGGCCGCAGGCCGCGCAGTTGGAGCCGGTGGGTATGTGGTCGAAGGCTGGGTAGCCGGTGGGCAATCCCTGTGGTCAAGCTCGTGGGCAGGCGCAGCCTGTCCATCAGCTTGTCCAGCAGGGTTGTCCACGGGCCGAGCGAAGCGAGCCAGCCGGTGGCCGC
terminator 2
Label Gene Product Position Strand
rrnB T1 terminator Escherichia coli rrnB 174..261 Fwd
DNA Sequence
CAAATAAAACGAAAGGCTCAGTCGAAAGACTGGGCCTTTCGTTTTATCTGTTGTTTGTCGGTGAACGCTCTCCTGAGTAGGACAAAT
rrnB T2 terminator 352..380 Fwd
DNA Sequence
AGAAGGCCATCCTGACGGATGGCCTTTT