mTol2-B-P-GFF-CR3 vector (V009838)

Basic Vector Information

      • Vector Name:
      • mTol2-B-P-GFF-CR3
      • Antibiotic Resistance:
      • Ampicillin
      • Length:
      • 6465 bp
      • Type:
      • Expression vector
      • Source/Author:
      • Otsuna H, Scoresby AN, Gaynes B, Tong ZZ, Kwan KK, Hutcheson DA, Chien CB, Dorsky R.

mTol2-B-P-GFF-CR3 vector Vector Map

mTol2-B-P-GFF-CR36465 bp300600900120015001800210024002700300033003600390042004500480051005400570060006300f1 oriAmpR promoterAmpRoriCAP binding sitelac promoterlac operatorM13 revSP6 promoterattB4attB1GAL4 DNA binding domainattB2attB3M13 fwdSV40 poly(A) signalSV40 poly(A) signalTagRFPcmlc2T7 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 ℃.

mTol2-B-P-GFF-CR3 vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       40924_2154        6465 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector mTol2-B-P-GFF-CR3, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6465)
  AUTHORS   Otsuna H, Scoresby AN, Gaynes B, Tong ZZ, Kwan KK, Hutcheson DA, 
            Chien CB, Dorsky R.
  TITLE     Large Scale Zebrafish Gal4 Enhancer Trap Screen for Neural Specific 
            Expression Patterns
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6465)
  AUTHORS   Otsuna H, Scoresby AN, Gaynes B, Tong ZZ, Kwan KK, Hutcheson DA, 
            Chien CB, Dorsky R.
  TITLE     Direct Submission
  JOURNAL   Submitted (19-MAR-2013) Neurobiology and Anatomy, University of 
            Utah, 20 North 1900 East, Salt Lake City, UT 84132, USA
REFERENCE   3  (bases 1 to 6465)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6465)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (19-MAR-2013) Neurobiology and Anatomy, University of Utah, 20 North
            1900 East, Salt Lake City, UT 84132, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..6465
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      181..636
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        714..818
                     /label=AmpR promoter
     CDS             819..1676
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      1850..2438
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    2726..2747
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        2762..2792
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2800..2816
                     /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)."
     primer_bind     2824..2840
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2858..2876
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     protein_bind    3144..3164
                     /label=attB4
                     /note="core recombination site for the Gateway(R) BP
                     reaction"
     protein_bind    3248..3272
                     /label=attB1
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3281..3721
                     /codon_start=1
                     /label=GAL4 DNA binding domain
                     /note="DNA binding domain of the GAL4 transcriptional
                     activator"
                     /translation="MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTK
                     RSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNK
                     DAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS"
     protein_bind    complement(3822..3846)
                     /label=attB2
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(4045..4065)
                     /label=attB3
                     /note="core recombination site for the Gateway(R) BP
                     reaction"
     primer_bind     complement(4073..4089)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    complement(4105..4239)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    4429..4563
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(4604..5314)
                     /codon_start=1
                     /label=TagRFP
                     /note="monomeric derivative of red fluorescent protein from
                     Entacmaea quadricolor (Merzlyak et al., 2007)"
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRSD
                     MALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAV
                     ARYCDLPSKLGHKLN"
     promoter        complement(5315..6213)
                     /label=cmlc2
                     /note="Cardiac myosin light chain 2 promoter"
     promoter        complement(join(6464..6465,1..17))
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"

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