pGBT-R14 vector (V006082)

Basic Vector Information

Vector Name:
pGBT-R14
Antibiotic Resistance:
Ampicillin
Length:
8392 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.
Promoter:
SP6

pGBT-R14 vector Map

pGBT-R148392 bp400800120016002000240028003200360040004400480052005600600064006800720076008000ol2loxPcarp beta-actin intron 1 splice acceptormRFP1Danio rerio beta-actin 3'UTR and poly(A) signalloxPol2T7 promoterM13 fwdf1 oriAmpR promoterAmpRoriCAP binding sitelac promoterlac operatorM13 revSP6 promoter

pGBT-R14 vector Sequence

LOCUS       40924_21200        8392 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pGBT-R14, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8392)
  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  JOURNAL   Unpublished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 8392)
  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 8392)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8392)
  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            "Unpublished"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..8392
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     mobile_element  complement(2..716)
                     /mobile_element_type="transposon:Tol2"
                     /label=ol2
                     /note="Tol2 long ITR-R"
     protein_bind    734..767
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     intron          811..908
                     /note="carp beta-actin intron 1 splice acceptor"
     CDS             941..1612
                     /label=mRFP1
                     /note="monomeric derivative of DsRed (Campbell et al.,
                     2002)"
     rep_origin      1588..2044
                     /label=F1 Ori
                     /note="F1 Ori"
     regulatory      1625..2629
                     /label=Danio rerio beta-actin 3'UTR and poly(A) signal
                     /note="Danio rerio beta-actin 3'UTR and poly(A) signal"
                     /regulatory_class="polyA_signal_sequence"
     misc_recomb     2726..2759
                     /label=loxP
                     /note="loxP"
     protein_bind    complement(2726..2759)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     mobile_element  2780..5453
                     /mobile_element_type="transposon:Tol2"
                     /label=ol2
                     /note="Tol2 long ITR-L"
     promoter        complement(5466..5484)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5491..5507)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      5648..6103
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        6181..6285
                     /label=AmpR promoter
     CDS             6286..7143
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      7317..7905
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    8193..8214
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        8229..8259
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8267..8283
                     /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     8291..8307
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        8325..8343
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"

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