Basic Vector Information
- Vector Name:
- pSFV4 (unavailable)
- Antibiotic Resistance:
- Ampicillin
- Length:
- 7834 bp
- Type:
- Expression vector
- Replication origin:
- ori
- Source/Author:
- Loppnau PA, Graslund S, Arrowsmith CH.
- Promoter:
- sacB
pSFV4 (unavailable) vector Vector Map
pSFV4 (unavailable) vector Sequence
LOCUS 40924_39827 7834 bp DNA circular SYN 18-DEC-2018
DEFINITION Expression vector pSFV4, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7834)
AUTHORS Loppnau PA, Graslund S, Arrowsmith CH.
TITLE A pipeline to generate recombinant antibodies to human target
proteins involved in epigenetic mechanisms
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 7834)
AUTHORS Loppnau PA, Graslund S, Arrowsmith CH.
TITLE Direct Submission
JOURNAL Submitted (14-NOV-2012) Structural Genomics Consortium, University
of Toronto, MaRS Centre, South tower, 101 College St, Room 700,
Toronto, Ontario M5G 1L7, Canada
REFERENCE 3 (bases 1 to 7834)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 7834)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Unpublished"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(14-NOV-2012) Structural Genomics Consortium, University of Toronto,
MaRS Centre, South tower, 101 College St, Room 700, Toronto, Ontario
M5G 1L7, Canada"
COMMENT SGRef: number: 3; type: "Journal Article"
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..7834
/mol_type="other DNA"
/organism="synthetic DNA construct"
rep_origin 12..465
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 492..596
/label=AmpR promoter
CDS 597..1454
/label=AmpR
/note="beta-lactamase"
rep_origin 1628..2216
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
misc_feature complement(2402..2544)
/label=bom
/note="basis of mobility region from pBR322"
CDS complement(2953..3126)
/label=lacZ-alpha
/note="LacZ-alpha fragment of beta-galactosidase"
protein_bind complement(3146..3162)
/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(3170..3200)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(3215..3236)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
CDS complement(3252..4331)
/label=lacI
/note="lac repressor"
promoter complement(4332..4409)
/label=lacIq promoter
/note="In the lacIq allele, a single base change in the
promoter boosts expression of the lacI gene about 10-fold."
protein_bind 4461..4482
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 4497..4527
/label=lac UV5 promoter
/note="E. coli lac promoter with an 'up' mutation"
protein_bind 4535..4551
/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 4606..4634
/label=tac promoter
/note="strong E. coli promoter; hybrid between the trp and
lac UV5 promoters"
protein_bind 4640..4660
/label=lac repressor protein binding site
/bound_moiety="lac repressor protein"
protein_bind 4642..4658
/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)."
regulatory 4701..4729
/label=Ptac
/note="Ptac"
/regulatory_class="promoter"
promoter 4701..4729
/label=tac promoter
/note="strong E. coli promoter; hybrid between the trp and
lac UV5 promoters"
regulatory 4701..4706
/label=derived from trp promoter
/note="derived from trp promoter"
/regulatory_class="minus_35_signal"
regulatory 4723..4729
/label=derived from lacUV5 promoter
/note="derived from lacUV5 promoter"
/regulatory_class="minus_10_signal"
protein_bind 4735..4755
/label=lac repressor protein binding site
/bound_moiety="lac repressor protein"
protein_bind 4737..4753
/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)."
sig_peptide 4796..4861
/label=pelB signal sequence
/note="leader peptide for secretion"
misc_feature 4862..5127
/note="partial sequence of FAB light chain constant region"
misc_feature 5113..5127
/label=5'-In-Fusion cloning site
/note="5'-In-Fusion cloning site"
protein_bind complement(5143..5176)
/label=loxP
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
promoter 5191..5636
/label=sacB promoter
/note="sacB promoter and control region"
CDS 5637..7055
/label=SacB
/note="secreted levansucrase that renders bacterial growth
sensitive to sucrose"
misc_feature 7120..7418
/note="partial coding sequence of FAB heavy chain constant
region"
misc_feature 7120..7134
/label=3'-In-Fusion cloning site
/note="3'-In-Fusion cloning site"
misc_feature 7419..7463
/label=C-terminal of FAB heavy chain with AviTag addition
/note="C-terminal of FAB heavy chain with AviTag addition"
CDS 7485..7529
/label=AviTag(TM)
/note="peptide tag that allows for enzymatic biotinylation"
misc_feature 7543..7587
/note="alternate C-terminal of FAB heavy chain without tag
addition"
misc_feature 7606..7644
/note="alternate C-terminal of FAB heavy chain with hexa
histidine-tag"
CDS 7654..7671
/label=6xHis
/note="6xHis affinity tag"
regulatory 7716..7743
/label=trp operon transcription termination sequence
/note="trp operon transcription termination sequence"
/regulatory_class="terminator"
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