Basic Vector Information
- Vector Name:
- pM14
- Antibiotic Resistance:
- Ampicillin
- Length:
- 5357 bp
- Type:
- CRIMIC donor vector
- Replication origin:
- ori
- Source/Author:
- Lee PT, Zirin J, Kanca O, Lin WW, Schulze KL, Li-Kroeger D, Tao R, Devereaux C, Hu Y, Chung V, Fang Y, He Y, Pan H, Ge M, Zuo Z, Housden BE, Mohr SE, Yamamoto S, Levis RW, Spradling AC, Perrimon N, Be
- Promoter:
- hsp70
pM14 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 ℃.
pM14 vector Sequence
LOCUS 40924_29596 5357 bp DNA circular SYN 18-DEC-2018
DEFINITION CRIMIC donor vector pM14, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5357)
AUTHORS Lee PT, Zirin J, Kanca O, Lin WW, Schulze KL, Li-Kroeger D, Tao R,
Devereaux C, Hu Y, Chung V, Fang Y, He Y, Pan H, Ge M, Zuo Z,
Housden BE, Mohr SE, Yamamoto S, Levis RW, Spradling AC, Perrimon N,
Bellen HJ.
TITLE A gene-specific T2A-GAL4 library for Drosophila
JOURNAL Elife 7, e35574 (2018)
PUBMED 29565247
REFERENCE 2 (bases 1 to 5357)
AUTHORS Lee P-T., Zirin J, Kanca O, Lin W-W., Schulze KL, Li-Kroeger D, Tao
R, Devereaux C, Hu Y, Chung V, Fang Y, He Y, Pan H, Ge M, Zuo Z,
Housden BE, Mohr SE, Yamamoto S, Levis RW, Spradling AC, Perrimon N,
Bellen HJ.
TITLE Direct Submission
JOURNAL
REFERENCE 3 (bases 1 to 5357)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 5357)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Elife 7,
e35574 (2018)"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(16-AUG-2018) HHMI, Molecular "
COMMENT SGRef: number: 3; type: "Journal Article"
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..5357
/mol_type="other DNA"
/organism="synthetic DNA construct"
rep_origin complement(3..458)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind 600..616
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 626..644
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 775..874
/label=phage phi-C31 attP
/note="attachment site of phage phi-C31"
protein_bind 931..964
/label=FRT (minimal)
/note="supports FLP-mediated excision but not integration
(Turan and Bode, 2011)"
misc_feature 971..1070
/label=MHC splice acceptor site
/note="MHC splice acceptor site"
regulatory 1071..1088
/label=stop codons in all reading frames
/note="stop codons in all reading frames"
/regulatory_class="terminator"
polyA_signal 1089..1223
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
regulatory 1093..1284
/label=SV40 polyadenylation signal
/note="SV40 polyadenylation signal"
/regulatory_class="polyA_signal_sequence"
regulatory 1331..1596
/label=3XP3 promoter
/note="3XP3 promoter"
/regulatory_class="promoter"
promoter 1602..1839
/label=hsp70 promoter
/note="Drosophila melanogaster hsp70Bb promoter"
CDS 1852..2568
/label=EGFP
/note="enhanced GFP"
polyA_signal complement(2572..2706)
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
misc_recomb 2814..2847
/label=FRT (minimal)
/note="FRT (minimal)"
protein_bind 2814..2847
/label=FRT (minimal)
/bound_moiety="FLP recombinase from the Saccharomyces
cerevisiae 2u plasmid"
/note="supports FLP-mediated excision but not integration
(Turan and Bode, 2011)"
protein_bind complement(2904..3003)
/label=phage phi-C31 attP
/note="attachment site of phage phi-C31"
primer_bind complement(3123..3139)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
promoter complement(3169..3187)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(3208..3224)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(3232..3248)
/label=lac operator
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(3256..3286)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(3301..3322)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(3610..4198)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(4372..5229)
/label=AmpR
/note="beta-lactamase"
promoter complement(5230..5334)
/label=AmpR promoter
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