pGBT-PX vector (V006083)

Basic Vector Information

Vector Name:
pGBT-PX
Antibiotic Resistance:
Ampicillin
Length:
8504 bp
Type:
Cloning vector
Replication origin:
ori
Source/Author:
Clark KJ, Balciunas D, Pagoda H-M., Ding Y, Westcot SE, Bedell VM, Greenwood TM, Urban MD, Skuster KJ, Petzold AM, Ni J, Nielson A, Sivasubbu S, Xu X, Hammerschmidt M, Ekker SC.

pGBT-PX vector Map

pGBT-PX8504 bp4008001200160020002400280032003600400044004800520056006000640068007200760080008400ol2loxPcarp beta-actin intron 1 splice acceptorT3 promotercarp beta-actin promoter regioncarp beta-actin exon 1 (non-coding)carp beta-actin intron 1 splice acceptorGFPcarp beta-actin intron 1 splice donorloxPol2T7 promoterM13 fwdf1 oriAmpR promoterAmpRoriCAP binding sitelac promoterlac operatorM13 revSP6 promoter

pGBT-PX vector Sequence

LOCUS       40924_21195        8504 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pGBT-PX, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8504)
  AUTHORS   Clark KJ, Balciunas D, Pagoda H-M., Ding Y, Westcot SE, Bedell VM, 
            Greenwood TM, Urban MD, Skuster KJ, Petzold AM, Ni J, Nielson A, 
            Sivasubbu S, Xu X, Hammerschmidt M, Ekker SC.
  TITLE     In vivo protein trapping produces a functional expression codex of 
            the vertebrate proteome
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8504)
  AUTHORS   Clark KJ, Balciunas D, Pagoda H-M., Ding Y, Westcot SE, Bedell VM, 
            Greenwood TM, Urban MD, Skuster KJ, Petzold AM, Ni J, Nielson A, 
            Sivasubbu S, Xu X, Hammerschmidt M, Ekker SC.
  TITLE     Direct Submission
  JOURNAL   Submitted (01-OCT-2010) Biochemistry and Molecular Biology, Mayo 
            Clinic, 221 4th Ave. SW, Rochester, MN 55902, USA
REFERENCE   3  (bases 1 to 8504)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8504)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (01-OCT-2010) Biochemistry and Molecular Biology, Mayo Clinic, 221 
            4th Ave. SW, Rochester, MN 55902, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8504
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     mobile_element  complement(1..192)
                     /mobile_element_type="transposon:Tol2"
                     /label=ol2
                     /note="mini ITR-R"
     misc_recomb     227..260
                     /label=loxP
                     /note="loxP"
     protein_bind    227..260
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     intron          313..410
                     /note="carp beta-actin intron 1 splice acceptor"
     promoter        1741..1759
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     regulatory      1768..2903
                     /label=carp beta-actin promoter region
                     /note="carp beta-actin promoter region"
                     /regulatory_class="promoter"
     exon            2909..3000
                     /note="carp beta-actin exon 1 (non-coding)"
     intron          3001..4264
                     /note="carp beta-actin intron 1 splice acceptor"
     CDS             4329..5042
                     /label=GFP
                     /note="green fluorescent protein"
     intron          5100..5236
                     /note="carp beta-actin intron 1 splice donor"
     protein_bind    complement(5254..5287)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     mobile_element  5304..5565
                     /mobile_element_type="transposon:Tol2"
                     /label=ol2
                     /note="mini ITR-L"
     promoter        complement(5578..5596)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5603..5619)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      5760..6215
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        6293..6397
                     /label=AmpR promoter
     CDS             6398..7255
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      7429..8017
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    8305..8326
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        8341..8371
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8379..8395
                     /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)."
     primer_bind     8403..8419
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        8437..8455
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"

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