LOCUS       Exported                5622 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pGBT-R16, complete sequence.
ACCESSION   HQ335169
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  JOURNAL   Unpublished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)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5622
                     /organism="Cloning vector pGBT-R16"
                     /mol_type="other DNA"
                     /db_xref="taxon:1001538"
     mobile_element  complement(1..192)
                     /mobile_element_type="transposon:Tol2"
                     /note="mini Tol2 ITR-R"
     misc_recomb     221..254
                     /label=loxP
                     /note="loxP"
     protein_bind    complement(221..254)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     intron          261..534
                     /note="Danio rerio beta-actin intron 3 splice acceptor"
     CDS             581..1255
                     /codon_start=1
                     /product="monomeric red fluorescent protein"
                     /label=monomeric red fluorescent protein
                     /note="mRFP; lacks start codon"
                     /protein_id="AEA29856.1"
                     /translation="ASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAKL
                     KVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGVV
                     TVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKMR
                     LKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHST
                     GA"
     regulatory      1265..2269
                     /regulatory_class="polyA_signal_sequence"
                     /note="Danio rerio beta-actin 3UTR and poly(A) signal"
     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"
                     /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 Ori"
     promoter        3411..3515
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3516..4376
                     /codon_start=1
                     /gene="ampR"
                     /product="beta-lactamase"
                     /label=ampR
                     /note="bla"
                     /protein_id="AEA29855.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            3516..4376
                     /gene="ampR"
                     /label=ampR
     rep_origin      complement(4437..5317)
                     /direction=LEFT
                     /label=ColE1 Ori
                     /note="ColE1 Ori"
     rep_origin      4547..5135
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    5423..5444
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /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
                     /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     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"