LOCUS       Exported                5035 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pPLV19_v2, complete sequence.
ACCESSION   KM062055
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5035)
  AUTHORS   Wendrich JR, Liao CY, van den Berg WA, De Rybel B, Weijers D.
  TITLE     Ligation-independent cloning for plant research
  JOURNAL   Methods Mol. Biol. 1284, 421-431 (2015)
  PUBMED    25757785
REFERENCE   2  (bases 1 to 5035)
  AUTHORS   Wendrich JR, Liao C-Y., van den Berg WAM., De Rybel B, Weijers D.
  TITLE     Direct Submission
  JOURNAL   Submitted (25-JUN-2014) Laboratory of Biochemistry, Wageningen 
            University, Dreijenlaan 3, Wageningen 6703HA, The Netherlands
REFERENCE   3  (bases 1 to 5035)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5035
                     /organism="Cloning vector pPLV19_v2"
                     /mol_type="other DNA"
                     /db_xref="taxon:1548286"
     rep_origin      complement(66..654)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      complement(719..5035)
                     /direction=LEFT
                     /note="ColE; high copy number E. coli origin of 
                     replication"
     CDS             complement(825..1640)
                     /codon_start=1
                     /product="nptI"
                     /label=nptI
                     /note="kanamycin resistance gene"
                     /protein_id="AIQ78276.1"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANVVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      1931..2366
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     rep_origin      1985..2401
                     /note="pSa ORI; pSa origin or replication"
     misc_feature    2501..2526
                     /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)"
     regulatory      complement(2532..2777)
                     /regulatory_class="terminator"
                     /note="tNOS; nopaline synthase terminator (tNOS)"
     terminator      2534..2786
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(2820..3371)
                     /codon_start=1
                     /product="BAR"
                     /label=BAR
                     /note="basta resistance gene"
                     /protein_id="AIQ78277.1"
                     /translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
                     WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
                     TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
                     WQLDFSLPVPPRPVLPVTEI"
     regulatory      complement(3372..3675)
                     /regulatory_class="promoter"
                     /note="pNOS; nopaline synthase (NOS) promoter"
     promoter        complement(3413..3592)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     primer_bind     3839..3855
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3865..3883
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     3909..3925
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    3946..3975
                     /note="LIC; ligation-independent cloning (LIC) site"
     CDS             3983..4702
                     /codon_start=1
                     /product="sCFP"
                     /label=sCFP
                     /note="super cyan fluorescent protein"
                     /protein_id="AIQ78278.1"
                     /translation="MTSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISDNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     regulatory      4755..4994
                     /regulatory_class="terminator"
                     /note="tNOS; nopaline synthase terminator (tNOS)"
     misc_feature    4986..5034
                     /note="RB; T-DNA right border"
     misc_feature    4986..5010
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"