LOCUS       Exported                5580 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pEnEOimCherryF3SGThsp, complete sequence.
ACCESSION   KF537341
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5580)
  AUTHORS   Koiwa H.
  TITLE     Arabidopsis thaliana CPL4 is an essential CTD-phosphatase
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5580)
  AUTHORS   Koiwa H.
  TITLE     Direct Submission
  JOURNAL   Submitted (13-AUG-2013) Department of Horticultural Science, Texas 
            A&M University, MS2133 TAMU, College Station, TX 77843-2133, USA
REFERENCE   3  (bases 1 to 5580)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..5580
                     /organism="Cloning vector pEnEOimCherryF3SGThsp"
                     /mol_type="other DNA"
                     /db_xref="taxon:1424654"
     rep_origin      complement(63..651)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(741..1550)
                     /codon_start=1
                     /product="KanR"
                     /label=KanR
                     /protein_id="AHA62522.1"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     protein_bind    1673..1772
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_recomb     1674..1769
                     /label=AttL2
                     /note="AttL2"
     regulatory      1798..2551
                     /regulatory_class="promoter"
                     /note="enhanced cauliflower mosaic virus 35S promoter EL2"
     promoter        2506..2551
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     regulatory      2555..2625
                     /regulatory_class="other"
                     /note="derived from Tobacco Mosaic Virus"
     intron          2642..3236
                     /note="derived from EF1a"
     CDS             3249..3956
                     /codon_start=1
                     /product="mCherry"
                     /label=mCherry
                     /protein_id="AHA62524.1"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     misc_feature    3963..4025
                     /label=3xFLAG tag
                     /note="3xFLAG tag"
     CDS             4026..4139
                     /codon_start=1
                     /product="streptavidin-binding peptide"
                     /label=SBP
                     /note="selected from a peptide library; binds streptavidin 
                     with nanomolar affinity (Keefe et al., 2001)"
                     /translation="MDEKTTGWRGGHVVEGLAGELEQLRARLEHHPQGQREP"
     CDS             4029..4568
                     /codon_start=1
                     /product="CTAP(SG)"
                     /label=CTAP
                     /protein_id="AHA62523.1"
                     /translation="DEKTTGWRGGHVVEGLAGELEQLRARLEHHPQGQREPIDENLYFQ
                     GIDENLYFQGGSTPAVTTYKLVINGKTLKGETTTKAVDAETAEKAFKQYANDNGVDGVW
                     TYDDATKTFTVTEVNTPAVTTYKLVINGKTLKGETTTKAVDAETAEKAFKQYANDNGVD
                     GVWTYDDATKTFTVTE"
     misc_feature    4146..4193
                     /label=TEV protease cleavage site
                     /note="TEV protease cleavage site"
     CDS             4146..4166
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             4173..4193
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     3UTR           4577..4736
                     /label=HSP18.2
                     /note="HSP18.2"
     regulatory      4737..4826
                     /regulatory_class="terminator"
                     /note="HSP18.2 terminator"
     terminator      4851..5103
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     protein_bind    complement(5124..5223)
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_recomb     complement(5127..5222)
                     /label=AttL1
                     /note="AttL1"
     terminator      5273..5300
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      5392..5478
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"