pM37(0) vector (V004854)

Basic Vector Information

Vector Name:
pM37(0)
Antibiotic Resistance:
Ampicillin
Length:
7829 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

pM37(0) vector Vector Map

pM37(0)7829 bp30060090012001500180021002400270030003300360039004200450048005100540057006000630066006900720075007800f1 oriM13 fwdT7 promoterphage phi-C31 attPFRT (minimal)MHC splice acceptor siteT2AGAL4SV40 polyadenylation signal3XP3 promoterEGFPSV40 poly(A) signalFRT (minimal)phage phi-C31 attPKS primerT3 promoterM13 revlac operatorlac promoterCAP binding siteoriAmpRAmpR promoter

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 ℃.

pM37(0) vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       40924_29606        7829 bp DNA     circular SYN 18-DEC-2018
DEFINITION  CRIMIC donor vector pM37(0), complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7829)
  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 7829)
  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 7829)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7829)
  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..7829
                     /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"
     CDS             1116..1169
                     /label=T2A
                     /note="2A peptide from Thosea asigna virus capsid protein"
     CDS             1170..3812
                     /label=GAL4
                     /note="GAL4 transcriptional activator"
     regulatory      3831..3965
                     /label=SV40 polyadenylation signal
                     /note="SV40 polyadenylation signal"
                     /regulatory_class="polyA_signal_sequence"
     polyA_signal    3831..3965
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     regulatory      4091..4296
                     /label=3XP3 promoter
                     /note="3XP3 promoter"
                     /regulatory_class="promoter"
     CDS             4324..5040
                     /label=EGFP
                     /note="enhanced GFP"
     polyA_signal    complement(5044..5178)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_recomb     5286..5319
                     /label=FRT (minimal)
                     /note="FRT (minimal)"
     protein_bind    5286..5319
                     /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(5376..5475)
                     /label=phage phi-C31 attP
                     /note="attachment site of phage phi-C31"
     primer_bind     complement(5595..5611)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(5641..5659)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5680..5696)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5704..5720)
                     /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(5728..5758)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5773..5794)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(6082..6670)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6844..7701)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(7702..7806)
                     /label=AmpR promoter

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