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krt4-sfGFP-tVE1 vector (V009892#)

Basic Vector Information

Basic Vector Information
Vector Name krt4-sfGFP-tVE1 Antibiotic Resistance Ampicillin
Length 8416 bp Type Cloning vector
Source Eskova A, Garcia-Junco R, Chauvigne F, Maischein H-M., Ammelburg M, Cerda J, Kaderali L, Nusslein-Volhard C, Irion U.

krt4-sfGFP-tVE1 vector Vector Map

krt4-sfGFP-tVE18416 bp4008001200160020002400280032003600400044004800520056006000640068007200760080008400M13 revattB4T7 promoterKS primerkrt4vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)SV40 poly(A) signalattB3M13 fwdSV40 poly(A) signalSK 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 ℃.

krt4-sfGFP-tVE1 vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported                8416 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector krt4-sfGFP-tVE1, complete sequence.
ACCESSION   KU645307
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8416)
  AUTHORS   Eskova A, Garcia-Junco R, Chauvigne F, Maischein H-M., Ammelburg M, 
            Cerda J, Kaderali L, Nusslein-Volhard C, Irion U.
  TITLE     Gain-of-function mutations of mau/DrAqp3a influence zebrafish 
            pigment pattern formation through the tissue environment
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8416)
  AUTHORS   Eskova A.
  TITLE     Direct Submission
  JOURNAL   Submitted (28-JAN-2016) e-CNV group, Max Planck Institute for 
            Developmental Biology, Spemannstr. 35-39, Tuebingen 72076, Germany
REFERENCE   3  (bases 1 to 8416)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8416
                     /organism="Cloning vector krt4-sfGFP-tVE1"
                     /mol_type="other DNA"
                     /label=derived from Danio rerio
                     /note="derived from Danio rerio"
                     /db_xref="taxon:1891373"
     primer_bind     1061..1077
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1097..1117
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     promoter        1119..1137
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     1163..1179
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      1195..3433
                     /regulatory_class="promoter"
                     /note="krt4"
     regulatory      3455..3464
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             3461..5083
                     /codon_start=1
                     /product="SfGFP/Aqp3aR220Q fusion protein"
                     /label=SfGFP/Aqp3aR220Q fusion protein
                     /note="tVE1 allele"
                     /protein_id="AOE47049.1"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIK
                     ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYKSGLRSMGWQKSVLDKLAQTFQIRNKLLRQGLAECLGTLILVM
                     FGCGSLAQLKLSEGSHGLFLTANLAFGFGATLGILVCGQVSGGHLNPAVTFALCLLGRE
                     KWRKFPVYFLFQTLGSFLGAAIIFAEYHDAIYDYAGESNELLVLGEKETAGIFATYPSK
                     YLTPLNGFFDQVIGTASLIVCILAIVDPYNNPIPQGLEAFTVGFSVLIIGLSMGFNSGY
                     AVNPAQDFGPRLFTAMAGWGSEVFTARDYWFLVPIFAPFIGAVIGVIVYQLMVGWHVEG
                     EARDKKAKAREEVMNLNDVASKE"
     polyA_signal    5116..5250
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    complement(5313..5333)
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     primer_bind     complement(5341..5357)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    5373..5507
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6179..6195)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(6232..6250)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6271..6287)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6295..6311
                     /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)."
     promoter        complement(6319..6349)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6364..6385
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(6673..7261)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7432..8292)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(8293..8397)
                     /gene="bla"
                     /label=AmpR promoter

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