TAL3393
PVT10497 2ug
TAL3393 Informaiton
Promoter: CMV promoter
Replicon: ori, F1 ori
Terminator: bGH poly (A) signal
Plasmid classification: gene library plasmid; other gene plasmids; fish gene plasmid.
Plasmid size: 8058bp
Plasmid tagging: N-3xFLAG, N-SV40 NLS, C-FokI cleavage domain
Prokaryotic resistance: ampicillin Amp (100 g/ml)
Screening marker: magnaporin blast
Clone strains: XL1 Blue and E. coli.
Culture conditions: 37 C, aerobic, LB
Expression host: mammalian cells such as 293T
Remarks: high copy plasmid
TAL3393 Description
TAL3393 is an eukaryotic expression plasmid expressing zebrafish gene. CMV promoter promotes the expression of Zebrafish Community-vasa-right gene.
TAL3393 Reference
Targeted gene disruption in somatic zebrafish cells using engineered TALENs.
TAL3393 Sequence
LOCUS Exported 8058 bp ds-DNA circular SYN 16-AUG-2017
KEYWORDS TAL3393
FEATURES Location/Qualifiers
source 1..8058
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 235..614
/note="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 615..818
/note="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
promoter 863..881
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 907..972
/codon_start=1
/product="three tandem FLAG(R) epitope tags, followed by an
enterokinase cleavage site"
/note="3xFLAG"
/translation="DYKDHDGDYKDHDIDYKDDDDK"
CDS 979..999
/codon_start=1
/locus_tag="
"
/product="nuclear localization signal of SV40 large T
antigen"
/note="SV40 NLS"
/note="
"
/protein_id="
"
/translation="PKKKRKV"
misc_feature 1020..3312
/note="ZebrafishCommunity-vasa-right"
CDS 3319..3906
/codon_start=1
/product="nonspecific DNA cleavage domain of the FokI
endonuclease (Li et al., 1992)"
/note="FokI cleavage domain"
/note="must dimerize to be catalytically active"
/protein_id="
"
/translation="QLVKSELEEKKSELRHKLKYVPHEYIELIEIARNSTQDRILEMKV
MEFFMKVYGYRGKHLGGSRKPDGAIYTVGSPIDYGVIVDTKAYSGGYNLPIGQADEMQR
YVEENQTRNKHINPNEWWKVYPSSVTEFKFLFVSGHFKGNYKAQLTRLNHITNCNGAVL
SVEELLIGGEMIKAGTLTLEEVRRKFNNGEINF"
polyA_signal 4019..4243
/note="bGH poly(A) signal"
/note="bovine growth hormone polyadenylation signal"
rep_origin 4289..4717
/direction=RIGHT
/note="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 4731..5061
/note="SV40 promoter"
/note="SV40 enhancer and early promoter"
rep_origin 4912..5047
/note="SV40 ori"
/note="SV40 origin of replication"
promoter 5109..5156
/note="EM7 promoter"
/note="synthetic bacterial promoter "
CDS 5175..5573
/codon_start=1
/gene="Aspergillus terreus BSD"
/product="blasticidin S deaminase"
/note="Blast"
/note="confers resistance to blasticidin"
/translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
KAIVKDSDGQPTAVGIRELLPSGYVWEG"
polyA_signal 5731..5852
/note="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
protein_bind 5925..5941
/bound_moiety="lac repressor encoded by lacI"
/note="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(5949..5979)
/note="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 5994..6015
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(6303..6891)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(7062..7922)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and