pGBT-RP2 vector (V006079)

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LOCUS       40924_21215        9119 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pGBT-RP2, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9119)
  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  JOURNAL   Unpublished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 9119)
  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 9119)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 9119)
  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            "Unpublished"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..9119
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    22..43
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        58..88
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    96..112
                     /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     120..136
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        154..172
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     mobile_element  complement(222..413)
                     /mobile_element_type="transposon:Tol2"
                     /label=ol2
                     /note="mini Tol2 ITR-R"
     misc_recomb     448..481
                     /label=loxP
                     /note="loxP"
     protein_bind    448..481
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     intron          488..620
                     /note="carp beta-actin intron 1 splice acceptor"
     CDS             653..1324
                     /label=mRFP1
                     /note="monomeric derivative of DsRed (Campbell et al.,
                     2002)"
     regulatory      1340..2515
                     /note="ocean pout antifreeze poly(A) signal; terminator; 
                     putative border element"
                     /regulatory_class="polyA_signal_sequence"
     rep_origin      1588..2044
                     /label=F1 Ori
                     /note="F1 Ori"
     promoter        2577..2595
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     regulatory      2604..3739
                     /label=carp beta-actin promoter region
                     /note="carp beta-actin promoter region"
                     /regulatory_class="promoter"
     exon            3745..3836
                     /note="carp beta-actin exon 1 (non-coding)"
     intron          3837..5100
                     /note="carp beta-actin intron 1"
     CDS             5165..5878
                     /label=GFP
                     /note="green fluorescent protein"
     intron          5945..6072
                     /note="carp beta-actin intron 1 splice donor"
     protein_bind    complement(6090..6123)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     mobile_element  6140..6401
                     /mobile_element_type="transposon:Tol2"
                     /label=ol2
                     /note="mini Tol2 ITR-L"
     promoter        complement(6414..6432)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6439..6455)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      6596..7051
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        7129..7233
                     /label=AmpR promoter
     CDS             7234..8091
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      8265..8853
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"