HITI-GmScreamM8-reconstituted-GFPNOS vector (V015431)

Basic Vector Information

Vector Name:
HITI-GmScreamM8-reconstituted-GFPNOS
Antibiotic Resistance:
Ampicillin
Length:
5290 bp
Type:
Cloning vector
Replication origin:
ori
Source/Author:
Gunadi A, Orchard N, Qu F, Finer J.

HITI-GmScreamM8-reconstituted-GFPNOS vector Vector Map

HITI-GmScreamM8-reconstituted-GFPNOS5290 bp6001200180024003000360042004800M13 fwdGmScreamM8 promoter from soybean5' HITI-1 junctionEGFPNOS terminator3' HITI-1 junctionM13 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 ℃.

HITI-GmScreamM8-reconstituted-GFPNOS vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       62056_1191        5290 bp DNA     circular SYN 01-FEB-2020
DEFINITION  Cloning vector HITI-GmScreamM8-reconstituted-GFPNOS, complete 
            sequence.
ACCESSION   MN043943
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5290)
  AUTHORS   Gunadi A, Orchard N, Qu F, Finer J.
  TITLE     Enhanced CRISPR/Cas9 mediated homology-independent targeted 
            integration in plant cotyledonary tissues
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5290)
  AUTHORS   Gunadi A, Orchard N, Qu F, Finer J.
  TITLE     Direct Submission
  JOURNAL   Submitted (06-JUN-2019) Horticulture and Crop Science, The Ohio 
            State University, 216 Williams Hall, 1680 Madison Ave., Wooster, OH 
            44691, USA
REFERENCE   3  (bases 1 to 5290)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (06-JUN-2019) Horticulture and Crop Science, The Ohio State 
            University, 216 Williams Hall, 1680 Madison Ave., Wooster, OH 44691,
            USA"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..5290
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      405..1978
                     /label=GmScreamM8 promoter from soybean
                     /note="GmScreamM8 promoter from soybean"
                     /regulatory_class="promoter"
     misc_feature    1979..2013
                     /label=5' HITI-1 junction
                     /note="5' HITI-1 junction"
     CDS             2014..2730
                     /label=EGFP
                     /note="enhanced GFP"
     terminator      2766..3018
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    3048..3059
                     /label=3' HITI-1 junction
                     /note="3' HITI-1 junction"
     primer_bind     complement(3069..3085)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3093..3109)
                     /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(3117..3147)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3162..3183)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(3471..4059)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4233..5090)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(5091..5195)
                     /label=AmpR promoter

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