LOCUS       40924_35224        5175 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pPLV20_v2, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5175)
  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 5175)
  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 5175)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5175)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Methods 
            Mol. Biol. 1284, 421-431 (2015)"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (25-JUN-2014) Laboratory of Biochemistry, Wageningen University, 
            Dreijenlaan 3, Wageningen 6703HA, The Netherlands"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5175
                     /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    2493..2515
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA
                     (truncated)"
     terminator      complement(2539..2791)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(2834..3625)
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     promoter        complement(3659..3838)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     primer_bind     3982..3998
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        4005..4023
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     4049..4065
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    4086..4115
                     /note="LIC; ligation-independent cloning (LIC) site"
     CDS             4123..4839
                     /label=SCFP3A
                     /note="monomeric optimized variant of ECFP (Kremers et al.,
                     2006)"
     terminator      4856..5104
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    5126..5150
                     /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 gagcgccttt gcccacaacc cggcggccgg ccgcaacaga
     2401 tcgttttata aatttttttt tttgaaaaag aaaaagcccg aaaggcggca acctctcggg
     2461 cttctggatt tccgatcccc ggaattagat cttggcagga tatattgtgg tgtaacgtta
     2521 tcagcttgca tgccggtcga tctagtaaca tagatgacac cgcgcgcgat aatttatcct
     2581 agtttgcgcg ctatattttg ttttctatcg cgtattaaat gtataattgc gggactctaa
     2641 tcataaaaac ccatctcata aataacgtca tgcattacat gttaattatt acatgcttaa
     2701 cgtaattcaa cagaaattat atgataatca tcgcaagacc ggcaacagga ttcaatctta
     2761 agaaacttta ttgccaaatg tttgaacgat ctgcttgact ctagctagag tccgaacccc
     2821 agagtcccgc tcagaagaac tcgtcaagaa ggcgatagaa ggcgatgcgc tgcgaatcgg
     2881 gagcggcgat accgtaaagc acgaggaagc ggtcagccca ttcgccgcca agctcttcag
     2941 caatatcacg ggtagccaac gctatgtcct gatagcggtc cgccacaccc agccggccac
     3001 agtcgatgaa tccagaaaag cggccatttt ccaccatgat attcggcaag caggcatcgc
     3061 cgtgggtcac gacgagatcc tcgccgtcgg gcatccgcgc cttgagcctg gcgaacagtt
     3121 cggctggcgc gagcccctga tgctcttcgt ccagatcatc ctgatcgaca agaccggctt
     3181 ccatccgagt acgtgctcgc tcgatgcgat gtttcgcttg gtggtcgaat gggcaggtag
     3241 ccggatcaag cgtatgcagc cgccgcattg catcagccat gatggatact ttctcggcag
     3301 gagcaaggtg agatgacagg agatcctgcc ccggcacttc gcccaatagc agccagtccc
     3361 ttcccgcttc agtgacaacg tcgagcacag ctgcgcaagg aacgcccgtc gtggccagcc
     3421 acgatagccg cgctgcctcg tcctgcagtt cattcagggc accggacagg tcggtcttga
     3481 caaaaagaac cgggcgcccc tgcgctgaca gccggaacac ggcggcatca gagcagccga
     3541 ttgtctgttg tgcccagtca tagccgaata gcctctccac ccaagcggcc ggagaacctg
     3601 cgtgcaatcc atcttgttca atcatgcctc gatcgagttg agagtgaata tgagactcta
     3661 attggatacc gaggggaatt tatggaacgt cagtggagca tttttgacaa gaaatatttg
     3721 ctagctgata gtgaccttag gcgacttttg aacgcgcaat aatggtttct gacgtatgtg
     3781 cttagctcat taaactccag aaacccgcgg ctcagtggct ccttcaacgt tgcggttctg
     3841 tcagttccaa acgtaaaacg gcttgtcccg cgtcatcggc gggggtcata acgtgactcc
     3901 cttaattctc cgctcatgat cgatgatgtg ctgcaaggcg attaagttgg gtaacgccag
     3961 ggttttccca gtcacgacgt tgtaaaacga cggccagtga attgtaatac gactcactat
     4021 agggcgaatt gggtaccggg ccccccctcg aggtcgacgg tatcgataag cttgatatcg
     4081 aattctagtt ggaatgggtt aacccaactc cataaggatc ccatgactag taagggcgag
     4141 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
     4201 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
     4261 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc
     4321 tggggcgtgc agtgcttcgc ccgctacccc gaccacatga agcagcacga cttcttcaag
     4381 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac
     4441 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
     4501 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
     4561 atcagcgaca acgtctatat caccgccgac aagcagaaga acggcatcaa ggccaacttc
     4621 aagatccgcc acaacatcga ggacggcggc gtgcagctcg ccgaccacta ccagcagaac
     4681 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc
     4741 aagctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
     4801 gccgccggga tcactctcgg catggacgag ctgtacaagt aaggatccga tccccgatcg
     4861 ttcaaacatt tggcaataaa gtttcttaag attgaatcct gttgccggtc ttgcgatgat
     4921 tatcatataa tttctgttga attacgttaa gcatgtaata attaacatgt aatgcatgac
     4981 gttatttatg agatgggttt ttatgattag agtcccgcaa ttatacattt aatacgcgat
     5041 agaaaacaaa atatagcgcg caaactagga taaattatcg cgcgcggtgt catctatgtc
     5101 tagagcggcc gccacgcgta ggccttgaca ggatatattg gcgggtaaac taagtcgctg
     5161 tatgtgtttg tttga
//
