Basic Vector Information
- Vector Name:
- pm!sp-gg
- Antibiotic Resistance:
- Gentamycin
- Length:
- 3192 bp
- Type:
- Cloning vector
- Replication origin:
- p15A ori
- Source/Author:
- Chavez A, Pruitt BW, Tuttle M, Shapiro RS, Cecchi RJ, Winston J, Turczyk BM, Tung M, Collins JJ, Church GM.
- Promoter:
- Pc
pm!sp-gg vector Vector Map
Plasmid Resuspension Protocol:
1. Centrifuge at 5,000×g for 5 min.
2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.
3. Close the tube and incubate for 10 minutes at room temperature.
4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.
5.Store the plasmid at -20 ℃.
pm!sp-gg vector Sequence
LOCUS 40924_29581 3192 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pm!sp-gg, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3192)
AUTHORS Chavez A, Pruitt BW, Tuttle M, Shapiro RS, Cecchi RJ, Winston J,
Turczyk BM, Tung M, Collins JJ, Church GM.
TITLE Precise Cas9 targeting enables genomic mutation prevention
JOURNAL Proc. Natl. Acad. Sci. U.S.A. (2018) In press
PUBMED 29555762
REFERENCE 2 (bases 1 to 3192)
AUTHORS Chavez A, Pruitt BW, Tuttle M, Shapiro RS, Cecchi RJ, Winston J,
Turczyk BM, Tung M, Collins JJ, Church GM.
TITLE Direct Submission
JOURNAL Submitted (02-MAR-2018) Synthetic Biology, Wyss Institute, 3
Blackfan Circle, Floor 5, Boston, MA 02115, USA
REFERENCE 3 (bases 1 to 3192)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 3192)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Proc. Natl.
Acad. Sci. U.S.A. (2018) In press"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(02-MAR-2018) Synthetic Biology, Wyss Institute, 3 Blackfan Circle,
Floor 5, Boston, MA 02115, USA"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..3192
/mol_type="other DNA"
/organism="synthetic DNA construct"
protein_bind 1..19
/label=tet operator
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 26..44
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
misc_feature 67..87
/label=golden gate cassette
/note="golden gate cassette"
misc_feature complement(67..73)
/label=SapI
/note="SapI"
misc_feature 74..80
/label=padding
/note="padding"
misc_feature 81..87
/label=SapI
/note="SapI"
repeat_region 88..123
/label=DR
/note="direct repeat for the Streptococcus pyogenes
CRISPR/Cas system"
CDS complement(988..1518)
/codon_start=1
/label=GmR
/note="gentamycin acetyltransferase"
/translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
EEVMHFDIDPSTAT"
promoter complement(1707..1735)
/label=Pc promoter
/note="class 1 integron promoter"
rep_origin complement(2379..2924)
/direction=LEFT
/label=p15A ori
/note="Plasmids containing the medium-copy-number p15A
origin of replication can be propagated in E. coli cells
that contain a second plasmid with the ColE1 origin."
This page is informational only.