LOCUS       Exported                9119 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pGBT-RP2, complete sequence.
ACCESSION   HQ335170
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  JOURNAL   Unpublished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)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9119
                     /organism="Cloning vector pGBT-RP2"
                     /mol_type="other DNA"
                     /db_xref="taxon:1001539"
     protein_bind    22..43
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /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
                     /bound_moiety="lac repressor encoded by lacI"
                     /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"
                     /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..1327
                     /codon_start=1
                     /product="monomeric red fluorescent protein"
                     /label=monomeric red fluorescent protein
                     /note="mRFP"
                     /protein_id="AEA29858.1"
                     /translation="ASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAKL
                     KVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGVV
                     TVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKMR
                     LKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHST
                     GA"
     regulatory      1340..2515
                     /regulatory_class="polyA_signal_sequence"
                     /note="ocean pout antifreeze poly(A) signal; terminator; 
                     putative border element"
     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
                     /regulatory_class="promoter"
                     /note="carp beta-actin promoter region"
     exon            3745..3836
                     /note="carp beta-actin exon 1 (non-coding)"
     intron          3837..5100
                     /note="carp beta-actin intron 1"
     CDS             5165..5881
                     /codon_start=1
                     /product="green fluorescent protein GM2 variant"
                     /label=green fluorescent protein GM2 variant
                     /note="GFP"
                     /protein_id="AEA29859.1"
                     /translation="MSKGEELFTGVVPILVELNGDVNGHKFPVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLAYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     intron          5945..6072
                     /note="carp beta-actin intron 1 splice donor"
     misc_recomb     6090..6123
                     /label=loxP
                     /note="loxP"
     protein_bind    6090..6123
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     mobile_element  6140..6401
                     /mobile_element_type="transposon:Tol2"
                     /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 Ori"
     promoter        7129..7233
                     /gene="bla"
                     /label=AmpR promoter
     CDS             7234..8094
                     /codon_start=1
                     /gene="ampR"
                     /product="beta-lactamase"
                     /label=ampR
                     /note="bla"
                     /protein_id="AEA29857.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            7234..8094
                     /gene="ampR"
                     /label=ampR
     rep_origin      complement(8155..9035)
                     /direction=LEFT
                     /label=ColE1 Ori
                     /note="ColE1 Ori"
     rep_origin      8265..8853
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"