LOCUS       40924_35094        7269 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pPLV04, complete sequence.
ACCESSION   .
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)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7269)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Plant 
            Physiol."; date: "2011"; volume: "156"; issue: "3"; pages: 
            "1292-1299"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (04-MAY-2011) Laboratory of Biochemistry, Wageningen University, 
            Dreijenlaan 3, Wageningen 6703HA, The Netherlands"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7269
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(66..654)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(828..1640)
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     rep_origin      1931..2366
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     misc_feature    2501..2523
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA
                     (truncated)"
     polyA_signal    complement(2531..2707)
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(2812..3603)
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     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             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
                     /label=EGFP
                     /note="enhanced GFP"
     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"
     terminator      6542..6790
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(6827..6845)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6866..6882)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(6890..6906)
                     /label=lac operator
                     /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    complement(6960..6981)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     misc_feature    7220..7244
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
ORIGIN
        1 gatctcatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
       61 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
      121 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
      181 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
      241 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
      301 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
      361 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
      421 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
      481 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
      541 gttaccttcg gaagaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
      601 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
      661 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
      721 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
      781 tttaaatcaa tctaaagtat atatgtgtaa cattggtcta gtgattagaa aaactcatcg
      841 agcatcaaat gaaactgcaa tttattcata tcaggattat caataccata tttttgaaaa
      901 agccgtttct gtaatgaagg agaaaactca ccgaggcagt tccataggat ggcaagatcc
      961 tggtatcggt ctgcgattcc gactcgtcca acatcaatac aacctattaa tttcccctcg
     1021 tcaaaaataa ggttatcaag tgagaaatca ccatgagtga cgactgaatc cggtgagaat
     1081 ggcaaaagtt tatgcatttc tttccagact tgttcaacag gccagccatt acgctcgtca
     1141 tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg attgcgcctg agcgagacga
     1201 aatacgcgat cgctgttaaa aggacaatta caaacaggaa tcgaatgcaa ccggcgcagg
     1261 aacactgcca gcgcatcaac aatattttca cctgaatcag gatattcttc taatacctgg
     1321 aatgctgttt tccctgggat cgcagtggtg agtaaccatg catcatcagg agtacggata
     1381 aaatgcttga tggtcggaag aggcataaat tccgtcagcc agtttagtct gaccatctca
     1441 tctgtaacaa cattggcaac gctacctttg ccatgtttca gaaacaactc tggcgcatcg
     1501 ggcttcccat acaatcggta gattgtcgca cctgattgcc cgacattatc gcgagcccat
     1561 ttatacccat ataaatcagc atccatgttg gaatttaatc gcggccttga gcaagacgtt
     1621 tcccgttgaa tatggctcat aacacccctt gtattactgt ttatgtaagc agacagtttt
     1681 attgttcatg atgatatatt tttatcttgt gcaatgtaac atcagagatt ttgagacaca
     1741 acgtggcttt gttgaataaa tcgaactttt gctgagttga aggatcagat cacgcatctt
     1801 cccgacaacg cagaccgttc cgtggcaaag caaaagttca aaatcaccaa ctggtccacc
     1861 tacaacaaag ctctcatcaa ccgtggctcc ctcactttct ggctggatga tggggcgatt
     1921 caggcgatcc ccatccaaca gcccgccgtc gagcgggctt ttttatcccc ggaagcctgt
     1981 ggatagaggg tagttatcca cgtgaaaccg ctaatgcccc gcaaagcctt gattcacggg
     2041 gctttccggc ccgctccaaa aactatccac gtgaaatcgc taatcagggt acgtgaaatc
     2101 gctaatcgga gtacgtgaaa tcgctaataa ggtcacgtga aatcgctaat caaaaaggca
     2161 cgtgagaacg ctaatagccc tttcagatca acagcttgca aacacccctc gctccggcaa
     2221 gtagttacag caagtagtat gttcaattag cttttcaatt atgaatatat atatcaatta
     2281 ttggtcgccc ttggcttgtg gacaatgcgc tacgcgcacc ggctccgccc gtggacaacc
     2341 gcaagcggtt gcccaccgtc gagcgccagc gcctttgccc acaacccggc ggccggccgc
     2401 aacagatcgt tttataaatt tttttttttg aaaaagaaaa agcccgaaag gcggcaacct
     2461 ctcgggcttc tggatttccg atccccggaa ttagagatct tggcaggata tattgtggtg
     2521 taacgttatc gatctggatt ttagtactgg attttggttt taggaattag aaattttatt
     2581 gatagaagta ttttacaaat acaaatacat actaagggtt tcttatatgc tcaacacatg
     2641 agcgaaaccc tataagaacc ctaatttccc ttatcgggaa actactcaca cattatttat
     2701 ggagaaaata gagagagata gatttgtaga gagagactgg tgatttcagc gtaccgaatt
     2761 gatcctctag ctagagtccg aaccccagag tcccgcacag agtcccgctc agaagaactc
     2821 gtcaagaagg cgatagaagg cgatgcgctg cgaatcggga gcggcgatac cgtaaagcac
     2881 gaggaagcgg tcagcccatt cgccgccaag ctcttcagca atatcacggg tagccaacgc
     2941 tatgtcctga tagcggtccg ccacacccag ccggccacag tcgatgaatc cagaaaagcg
     3001 gccattttcc accatgatat tcggcaagca ggcatcgccc tgggtcacga cgagatcctc
     3061 gccgtcgggc atccgcgcct tgagcctggc gaacagttcg gctggcgcga gcccctgatg
     3121 ctcttcgtcc agatcatcct gatcgacaag accggcttcc atccgagtac gtcctcgctc
     3181 gatgcgatgt ttcgcttggt ggtcgaatgg gcaggtagcc ggatcaagcg tatgcagccg
     3241 ccgcattgca tcagccatga tggatacttt ctcggcagga gcaaggtgag atgacaggag
     3301 atcctgcccc ggcacttcgc ccaatagcag ccagtccctt cccgcttcag tgacaacgtc
     3361 gagcacagct gcgcaaggaa cgcccgtcgt ggccagccac gatagccgcg ctgcctcgtc
     3421 ttggagttca ttcagggcac cggacaggtc ggtcttgaca aaaagaaccg ggcgcccctg
     3481 cgctgacagc cggaacacgg cggcatcaga gcagccgatt gtctgttgtg cccagtcata
     3541 gccgaatagc ctctccaccc aagcggccgg agaacctgcg tgcaatccat cttgttcaat
     3601 catcttgttc aatcatgcca gcttgggctg tcctctccaa atgaaatgaa cttccttata
     3661 tagaggaagg gtcttgcgaa ggatagtggg attgtgcgtc atcccttacg tcagtggaga
     3721 tgtcacatca atccacttgc tttgaagacg tggttggaac gtcttctttt tccacgatgc
     3781 tcctcgtggg tgggggtcca tctttgggac cactgtcggc agaggcatct tgaatgatag
     3841 cctttccttt atcgcaatga tggcatttgt aggagccacc ttccttttct actgtccttt
     3901 cgatgaagtg acagatagct gggcaatgga atccgaggag gtttcccgaa attacccttt
     3961 gttgaaaagt ctcaatagcc ctttggtctt ctgagactgt atctttgaca tttttggagt
     4021 aggggtacga taacattaac gtttacaatt tcgcgccatt cgccattcag gctgcgcaac
     4081 tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga
     4141 tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa
     4201 acgacggcca gtgaattgta atacgactca ctatagggcg aattgggtac cgggcccccc
     4261 ctcgaggtcg acggtatcga taagcttgat atcgaattct agttggaatg ggttaaccca
     4321 actccataag gatccatggc tccaaagaag aagagaaagg tcatggtgag caagggcgag
     4381 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
     4441 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
     4501 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc
     4561 tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag
     4621 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac
     4681 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
     4741 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
     4801 aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc
     4861 aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
     4921 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc
     4981 gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
     5041 gccgccggga tcactctcgg catggacgag ctgtacaaga gatccatggt gagcaagggc
     5101 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
     5161 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg
     5221 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg
     5281 acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc
     5341 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc
     5401 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
     5461 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
     5521 tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac
     5581 ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag
     5641 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag
     5701 tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
     5761 accgccgccg ggatcactct cggcatggac gagctgtaca agagatccat ggtgagcaag
     5821 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
     5881 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc
     5941 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
     6001 ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc
     6061 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
     6121 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     6181 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     6241 aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg
     6301 aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag
     6361 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc
     6421 cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
     6481 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtaaag atccgatccc
     6541 cgatcgttca aacatttggc aataaagttt cttaagattg aatcctgttg ccggtcttgc
     6601 gatgattatc atataatttc tgttgaatta cgttaagcat gtaataatta acatgtaatg
     6661 catgacgtta tttatgagat gggtttttat gattagagtc ccgcaattat acatttaata
     6721 cgcgatagaa aacaaaatat agcgcgcaaa ctaggataaa ttatcgcgcg cggtgtcatc
     6781 tatgtctaga gcggccgcca ccgcggtgga gctccagctt ttgttccctt tagtgagggt
     6841 taattccgag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     6901 tcacaattcc acacaacata cgagccggaa ghcataaagt gtaaagcctg gggtgcctaa
     6961 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     7021 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     7081 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     7141 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     7201 ggaaagaaca tgaaggcctt gacaggatat attggcgggt aaactaagtc gctgtatgtg
     7261 tttgtttga
//
