LOCUS       Exported                4686 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pEnEOTG-stop, complete sequence.
ACCESSION   KF545096
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4686)
  AUTHORS   Koiwa H.
  TITLE     Arabidopsis CPL4 is an essential CTD phosphatase
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4686)
  AUTHORS   Koiwa H.
  TITLE     Direct Submission
  JOURNAL   Submitted (15-AUG-2013) Department of Horticultural Science, Texas 
            A&M University, MS2133 TAMU, College Station, TX 77843-2133, USA
REFERENCE   3  (bases 1 to 4686)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..4686
                     /organism="Cloning vector pEnEOTG-stop"
                     /mol_type="other DNA"
                     /db_xref="taxon:1427726"
     regulatory      24..776
                     /regulatory_class="promoter"
                     /note="enhanced cauliflower mosaic virus 35S promoter EL2"
     promoter        728..773
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     misc_feature    777..852
                     /label=Tobacco mosaic virus omega
                     /note="Tobacco mosaic virus omega"
     CDS             859..2202
                     /codon_start=1
                     /product="N-TAP sGFP"
                     /label=N-TAP sGFP
                     /protein_id="AHA91198.1"
                     /translation="MAGLAQHDEAVDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLK
                     DDPSQSANLLAEAKKLNDAQAPKVDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKD
                     DPSQSANLLAEAKKLNDAQAPKVDANCDIPTTASENLYFQGELKRRWKKNFIAVSAANR
                     FKKISSSGALDDDDKLIDTVDLEVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGD
                     ATYGKLTLKFICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQE
                     RTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMA
                     DKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNE
                     KRDHMVLLEFVTAAGITHGMDELYKTREIERPPPRWSSNFPDRSNIWQ"
     CDS             889..1062
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLL
                     AEAKKLNDAQAPK"
     CDS             1066..1236
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="DNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLLA
                     EAKKLNDAQAPK"
     CDS             1273..1293
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             1300..1377
                     /codon_start=1
                     /product="calmodulin-binding peptide"
                     /label=CBP
                     /note="derived from skeletal muscle myosin light chain 
                     kinase; binds calmodulin with nanomolar affinity in the 
                     presence of calcium"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     CDS             1378..1392
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
     primer_bind     complement(1398..1414)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             1417..2127
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELY"
     regulatory      2169..2439
                     /regulatory_class="terminator"
                     /note="Nos terminator"
     terminator      2179..2431
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_recomb     2452..2551
                     /label=AttL1
                     /note="AttL1"
     protein_bind    complement(2452..2551)
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     terminator      2601..2628
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      2720..2806
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(2912..3585)
                     /direction=LEFT
                     /label=pUC origin
                     /note="pUC origin"
     rep_origin      complement(2971..3559)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3649..4458)
                     /codon_start=1
                     /product="KanR"
                     /label=KanR
                     /protein_id="AHA91197.1"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     misc_recomb     4581..4680
                     /label=AttL2
                     /note="AttL2"
     protein_bind    4581..4680
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"