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mTol2-B-P-GFF-CG3 vector (V009839#)

Basic Vector Information

Basic Vector Information
Vector Name mTol2-B-P-GFF-CG3 Antibiotic Resistance Ampicillin
Length 6411 bp Type Expression vector
Source Otsuna H, Scoresby AN, Gaynes B, Tong ZZ, Kwan KK, Hutcheson DA, Chien CB, Dorsky R.

mTol2-B-P-GFF-CG3 vector Vector Map

mTol2-B-P-GFF-CG36411 bp300600900120015001800210024002700300033003600390042004500480051005400570060006300M13 fwdf1 oriAmpR promoterampRcolE1 oriCAP binding sitelac promoterlac operatorM13RSP6 promoterTol2 3' TIRM13RattB4adenovirus E1B TATA5'UTRattB1Gal4FFattB2SV40 late polyA from pCS2+attB3M13 fwdSV40 late polyA from pCS2+SV40 late polyA (from pCS2+)T7 promoterEGFPcmlc2 promoterTol2 5' TIR

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-CG3 vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported                6411 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector mTol2-B-P-GFF-CG3, complete sequence.
ACCESSION   KC800698
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6411)
  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  JOURNAL   Unpublished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 6411)
  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 6411)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..6411
                     /organism="Expression vector mTol2-B-P-GFF-CG3"
                     /mol_type="other DNA"
                     /db_xref="taxon:1344357"
     primer_bind     complement(24..40)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(25..40)
                     /label=M13F
                     /note="M13F"
     rep_origin      181..636
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     rep_origin      198..504
                     /label=f1(+) ori
                     /note="f1(+) ori"
     promoter        714..818
                     /gene="bla"
                     /label=AmpR promoter
     CDS             819..1679
                     /codon_start=1
                     /product="ampR"
                     /label=ampR
                     /protein_id="AGN90798.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      1774..2456
                     /label=colE1 ori
                     /note="colE1 ori"
     rep_origin      1824..2497
                     /label=pUC ori
                     /note="pUC ori"
     rep_origin      1850..2438
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    2726..2747
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /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
                     /bound_moiety="lac repressor encoded by lacI"
                     /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..2845
                     /label=M13R
                     /note="M13R"
     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"
     regulatory      2858..2874
                     /regulatory_class="promoter"
                     /note="SP6"
     repeat_region   2927..2943
                     /label=Tol2 3' TIR
                     /note="Tol2 3' TIR"
     primer_bind     3108..3129
                     /label=M13R
                     /note="M13R"
     primer_bind     3108..3124
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_recomb     3144..3164
                     /label=attB4
                     /note="attB4"
     misc_feature    3144..3164
                     /label=att4_shared
                     /note="att4_shared"
     protein_bind    3144..3164
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     regulatory      3193..3203
                     /regulatory_class="promoter"
                     /note="adenovirus E1B TATA"
     5'UTR           3211..3242
                     /label=carp beta-actin trx start
                     /note="carp beta-actin trx start"
     protein_bind    3248..3272
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_recomb     3249..3255
                     /label=attB1
                     /note="attB1"
     misc_feature    3253..3255
                     /label=att1_shared
                     /note="att1_shared"
     misc_recomb     3256..3272
                     /label=attL1
                     /note="attL1"
     misc_feature    3256..3266
                     /label=att1_shared
                     /note="att1_shared"
     misc_feature    3281..3820
                     /label=Gal4FF
                     /note="Gal4FF"
     CDS             3281..3721
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="DNA binding domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 DNA binding domain
                     /translation="MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTK
                     RSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNK
                     DAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS"
     misc_feature    3281..3720
                     /label=Gal4 DBD
                     /note="Gal4 DBD"
     protein_bind    complement(3822..3846)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_recomb     3822..3843
                     /label=attL2
                     /note="attL2"
     misc_feature    3828..3843
                     /label=att2_shared
                     /note="att2_shared"
     misc_recomb     3844..3845
                     /label=attB2
                     /note="attB2"
     misc_feature    3848..4043
                     /label=SV40 late polyA from pCS2+
                     /note="SV40 late polyA from pCS2+"
     polyA_signal    3848..3982
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_recomb     4045..4065
                     /label=attB3
                     /note="attB3"
     protein_bind    complement(4045..4065)
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     misc_feature    4045..4059
                     /label=att3_shared
                     /note="att3_shared"
     primer_bind     complement(4073..4089)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4074..4089)
                     /label=M13F
                     /note="M13F"
     misc_feature    4105..4302
                     /label=SV40 late polyA from pCS2+
                     /note="SV40 late polyA from pCS2+"
     polyA_signal    4105..4239
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    complement(4361..4493)
                     /label=SV40 late polyA (from pCS2+)
                     /note="SV40 late polyA (from pCS2+)"
     promoter        4498..4515
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(4541..5260)
                     /codon_start=1
                     /product="EGFP"
                     /label=EGFP
                     /protein_id="AGN90797.1"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     misc_feature    complement(5056..5069)
                     /label=EGFP-like chromophore
                     /note="EGFP-like chromophore"
     regulatory      complement(5261..6159)
                     /regulatory_class="promoter"
                     /note="cmlc2 promoter"
     repeat_region   6379..6397
                     /label=Tol2 5' TIR
                     /note="Tol2 5' TIR"
     primer_bind     complement(6407..18)
                     /label=T7 primer
                     /note="T7 primer"
     promoter        complement(6410..17)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"

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