pAcCAST-sgRNA_entry (CJT83)

Plasmid Information

Purpose

Expresses AcCAST. New sgRNA spacer sequences can be added using Golden Gate assembly (via SapI sites).

Depositor

Benjamin Kleinstiver

Article

Tou et al Nat Biotechnol. 2023 Jan 2. doi: 10.1038

Insert

AcTnsB, AcTnsC, AcTniQ, AcCas12k

Promoter

Lac and J23119

Available Since

Feb. 18, 2022

Availability

Available

Length

8,024 bp

Topology

circular

Molecule Type

other DNA

CDS 1

Label	Gene	Product	Position	Strand
AmpR	bla	beta-lactamase	947..1808	Rev
DNA Sequence ATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAA Protein Sequence MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW

promoter 5

Label	Gene	Product	Position	Strand
AmpR promoter	bla	—	1808..1913	Rev
DNA Sequence CGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGT
BBa_J23108	—	—	2261..2296	Rev
DNA Sequence CTGACAGCTAGCTCAGTCCTAGGTATAATGCTAGC
lac promoter	—	—	4346..4377	Rev
DNA Sequence TTTACACTTTATGCTTCCGGCTCGTATGTTG
lac promoter	—	—	7736..7767	Rev
DNA Sequence TTTACACTTTATGCTTCCGGCTCGTATGTTG
lac promoter	—	—	7865..7896	Rev
DNA Sequence TTTACACTTTATGCTTCCGGCTCGTATGTTG

protein_bind 2

Label	Gene	Product	Position	Strand
lac operator	—	—	7841..7858	Fwd
DNA Sequence TTGTTATCCGCTCACAA
CAP binding site	—	—	7910..7932	Fwd
DNA Sequence ATGAGTGAGCTAACTCACATTA

rep_origin 1

Label	Gene	Product	Position	Strand
ori	—	—	188..777	Rev
DNA Sequence TTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTTCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAA

terminator 2

Label	Gene	Product	Position	Strand
T7Te terminator	—	—	1939..1967	Rev
DNA Sequence GGCTCACCTTCGGGTGGGCCTTTCTGCG
rrnB T1 terminator	Escherichia coli rrnB	—	2309..2381	Rev
DNA Sequence CAAATAAAACGAAAGGCTCAGTCGAAAGACTGGGCCTTTCGTTTTATCTGTTGTTTGTCGGTGAACGCTCTC