pSFV4 (unavailable) vector (V003281)

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)7834 bp30060090012001500180021002400270030003300360039004200450048005100540057006000630066006900720075007800f1 oriAmpR promoterAmpRoribomlacZ-alphalac operatorlac promoterCAP binding sitelacIlacIq promoterCAP binding sitelac UV5 promoterlac operatortac promoterlac repressor protein binding sitePtaclac repressor protein binding sitepelB signal sequencepartial sequence of FAB light chain constant regionloxPsacB promoterSacBpartial coding sequence of FAB heavy chain constant regionC-terminal of FAB heavy chain with AviTag additionAviTag(TM)alternate C-terminal of FAB heavy chain without tag additionalternate C-terminal of FAB heavy chain with hexa histidine-tag6xHistrp operon transcription termination sequence

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  JOURNAL   Unpublished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            "Unpublished"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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