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 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TITLE A pipeline to generate recombinant antibodies to human target proteins involved in epigenetic mechanisms JOURNAL Unpublished REFERENCE 2 (bases 1 to 7834) AUTHORS Loppnau PA, Graslund S, Arrowsmith CH.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 TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Unpublished" COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted (14-NOV-2012) Structural Genomics Consortium, University of Toronto, MaRS Centre,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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