LOCUS       Exported                7269 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pPLV04, complete sequence.
ACCESSION   JF909457
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7269)
  AUTHORS   De Rybel B, van den Berg W, Lokerse A, Liao CY, van Mourik H, Moller
            B, Peris CL, Weijers D.
  TITLE     A versatile set of ligation-independent cloning vectors for 
            functional studies in plants
  JOURNAL   Plant Physiol. 156 (3), 1292-1299 (2011)
  PUBMED    21562332
REFERENCE   2  (bases 1 to 7269)
  AUTHORS   De Rybel B, van den Berg W, Weijers D.
  TITLE     Direct Submission
  JOURNAL   Submitted (04-MAY-2011) Laboratory of Biochemistry, Wageningen 
            University, Dreijenlaan 3, Wageningen 6703HA, The Netherlands
REFERENCE   3  (bases 1 to 7269)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7269
                     /organism="Cloning vector pPLV04"
                     /mol_type="other DNA"
                     /db_xref="taxon:1035711"
     rep_origin      complement(66..654)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(825..1640)
                     /codon_start=1
                     /product="KanR"
                     /label=KanR
                     /note="kanamycin resistance"
                     /protein_id="AEG42738.1"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANVVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      1931..2366
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     rep_origin      1985..2480
                     /note="pSa ORI; pSa Origin or Replication"
     misc_feature    2501..2527
                     /note="LB; T-DNA Left Border"
     misc_feature    2501..2523
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
     polyA_signal    2531..2707
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     regulatory      complement(2534..2808)
                     /regulatory_class="terminator"
                     /note="t35S; CaMV 35S terminator"
     CDS             complement(2809..3603)
                     /codon_start=1
                     /product="NPTII"
                     /label=NPTII
                     /note="NPTII kanamycin resistance"
                     /protein_id="AEG42739.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERGRTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTQGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     regulatory      complement(3604..4092)
                     /regulatory_class="promoter"
                     /note="p35S; CaMV 35S promoter"
     promoter        complement(3629..3974)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     4176..4192
                     /note="M13F; M13 forward primer site"
     primer_bind     4196..4212
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        4219..4237
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     4263..4279
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    4300..4329
                     /note="LIC; ligation-independent cloning (LIC) site"
     CDS             4336..6528
                     /codon_start=1
                     /product="SV40-3xeGFP"
                     /label=SV40-3xeGFP
                     /note="SV40 nuclear localization signal fused N-terminally 
                     to tandem trimeric enhanced green fluorescent protein 
                     (3xeGFP)"
                     /protein_id="AEG42740.1"
                     /translation="MAPKKKRKVMVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEG
                     DATYGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQ
                     ERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIM
                     ADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPN
                     EKRDHMVLLEFVTAAGITLGMDELYKRSMVSKGEELFTGVVPILVELDGDVNGHKFSVS
                     GEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMP
                     EGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSH
                     NVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSAL
                     SKDPNEKRDHMVLLEFVTAAGITLGMDELYKRSMVSKGEELFTGVVPILVELDGDVNGH
                     KFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFF
                     KSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEY
                     NYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLS
                     TQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYK"
     CDS             4342..4362
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4363..5079
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             5086..5802
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             5809..6528
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     regulatory      6535..6784
                     /regulatory_class="terminator"
                     /note="tNOS; nopaline synthase terminator (tNOS)"
     promoter        complement(6827..6845)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6865..6882)
                     /note="M13R; M13 reverse primer site"
     primer_bind     complement(6866..6882)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6890..6906
                     /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)."
     promoter        complement(6914..6945)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6960..6981
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     misc_feature    7220..7268
                     /note="RB; T-DNA Right Border"
     misc_feature    7220..7244
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"