pGAD424A20ZF- vector (V006120)

Basic Vector Information

Vector Name:
pGAD424A20ZF-
Antibiotic Resistance:
Ampicillin
Length:
7804 bp
Type:
Yeast expression vector
Replication origin:
ori
Host:
Yeast
Source/Author:
De Schamphelaire W, Olbrechts A, Meert J, Verhelst K, Roggeman Fonseca M, Vanhoucke M, Beyaert R.
Promoter:
LEU2

pGAD424A20ZF- vector Map

pGAD424A20ZF-7804 bp30060090012001500180021002400270030003300360039004200450048005100540057006000630066006900720075007800ADH1 promoterSV40 NLSGAL4 activation domainHAunnamed protein product; hA20fADH1 terminatorLEU2LEU2 promoterM13 revlac operatorlac promoterCAP binding siteoriAmpRAmpR promoter2u ori

pGAD424A20ZF- vector Sequence

LOCUS       40924_20940        7804 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Yeast expression vector pGAD424A20ZF-, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7804)
  AUTHORS   De Schamphelaire W, Olbrechts A, Meert J, Verhelst K, Roggeman 
            Fonseca M, Vanhoucke M, Beyaert R.
  TITLE     BCCM/LMBP Plasmid collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7804)
  AUTHORS   De Schamphelaire W.
  TITLE     Direct Submission
  JOURNAL   Submitted (02-FEB-2017) BCCM/LMBP, Universiteit Gent, 
            Technologiepark 927, 9052, BELGIUM
REFERENCE   3  (bases 1 to 7804)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7804)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (02-FEB-2017) BCCM/LMBP, Universiteit Gent, Technologiepark 927, 
            9052, BELGIUM"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7804
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        5..406
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             452..472
                     /label=SV40 NLS
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
     CDS             488..829
                     /label=GAL4 activation domain
                     /note="activation domain of the GAL4 transcriptional
                     activator"
     CDS             845..871
                     /label=HA
                     /note="HA (human influenza hemagglutinin) epitope tag"
     CDS             887..1987
                     /codon_start=1
                     /note="unnamed protein product; hA20f"
                     /protein_id="SJL88290.1"
                     /translation="MAEQVLPQALYLSNMRKAVKIRERTPEDIFKPTNGIIHHFKTMHR
                     YTLEMFRTCQFCPQFREIIHKALIDRNIQATLESQKKLNWCREVRKLVALKTNGDGNCL
                     MHATSQYMWGVQDTDLVLRKALFSTLKETDTRNFKFRWQLESLKSQEFVETGLCYDTRN
                     WNDEWDNLIKMASTDTPMARSGLQYNSLEEIHIFVLCNILRRPIIVISDKMLRSLESGS
                     NFAPLKVGGIYLPLHWPAQECYRYPIVLGYDSHHFVPLVTLKDSGPEIRAVPLVNRDRG
                     RFEDLKVHFLTDPENEMKEKLLKEYLMVIEIPVQGWDHGTTHLINAAKLDEANLPKEIN
                     LVDDYFELVQHEYKKWQENSEQGRREG"
     terminator      2386..2573
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae
                     alcohol dehydrogenase 1 (ADH1) gene"
     CDS             complement(2694..3785)
                     /label=LEU2
                     /note="3-isopropylmalate dehydrogenase, required for
                     leucine biosynthesis"
     promoter        complement(3786..4190)
                     /label=LEU2 promoter
     primer_bind     complement(4232..4248)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4256..4272)
                     /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(4280..4310)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4325..4346)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4634..5222)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5396..6253)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(6254..6358)
                     /label=AmpR promoter
     rep_origin      6640..7804
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"

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