pGBT-R16 vector (V006080)

Basic Vector Information

Vector Name:
pGBT-R16
Antibiotic Resistance:
Ampicillin
Length:
5622 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-R16 vector Map

pGBT-R165622 bp60012001800240030003600420048005400ol2loxPDanio rerio beta-actin intron 3 splice acceptormRFP1Danio rerio beta-actin 3'UTR and poly(A) signalloxPol2T7 promoterM13 fwdf1 oriAmpR promoterAmpRoriCAP binding sitelac promoterlac operatorM13 revSP6 promoter

pGBT-R16 vector Sequence

LOCUS       40924_21210        5622 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pGBT-R16, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5622)
  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  JOURNAL   Unpublished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 5622)
  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 5622)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5622)
  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            "Unpublished"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..5622
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     mobile_element  complement(1..192)
                     /mobile_element_type="transposon:Tol2"
                     /label=ol2
                     /note="mini Tol2 ITR-R"
     protein_bind    221..254
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     intron          261..534
                     /note="Danio rerio beta-actin intron 3 splice acceptor"
     CDS             581..1252
                     /label=mRFP1
                     /note="monomeric derivative of DsRed (Campbell et al.,
                     2002)"
     regulatory      1265..2269
                     /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"
     rep_origin      1588..2044
                     /label=F1 Ori
                     /note="F1 Ori"
     misc_recomb     2366..2399
                     /label=loxP
                     /note="loxP"
     protein_bind    complement(2366..2399)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     mobile_element  2422..2683
                     /mobile_element_type="transposon:Tol2"
                     /label=ol2
                     /note="mini Tol2 ITR-L"
     promoter        complement(2696..2714)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2721..2737)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      2878..3333
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3411..3515
                     /label=AmpR promoter
     CDS             3516..4373
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      4547..5135
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    5423..5444
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        5459..5489
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5497..5513
                     /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     5521..5537
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        5555..5573
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"

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