LOCUS       40924_10991        4924 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Binary vector pCLEAN-G144, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4924)
  AUTHORS   Thole V, Worland B, Snape JW, Vain P.
  TITLE     The pCLEAN dual binary vector system for Agrobacterium-mediated 
            plant transformation
  JOURNAL   Plant Physiol. 145 (4), 1211-1219 (2007)
  PUBMED    17932303
REFERENCE   2  (bases 1 to 4924)
  AUTHORS   Thole V, Vain P.
  TITLE     Direct Submission
  JOURNAL   Submitted (01-OCT-2007) Department of Crop Genetics, John Innes 
            Centre, Colney Lane, Norwich NR4 7UH, UK
REFERENCE   3  (bases 1 to 4924)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4924)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Plant 
            Physiol."; date: "2007"; volume: "145"; issue: "4"; pages: 
            "1211-1219"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (01-OCT-2007) Department of Crop Genetics, John Innes Centre, Colney
            Lane, Norwich NR4 7UH, UK"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4924
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(61..649)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(823..1635)
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     regulatory      complement(1774..2070)
                     /label=soybean-derived poly(A)signal
                     /note="soybean-derived poly(A)signal"
                     /regulatory_class="terminator"
     CDS             complement(2106..2816)
                     /label=mgfp5
                     /note="GFP with folding enhancement mutations"
     promoter        complement(2915..3259)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     rep_origin      3556..3991
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     misc_feature    4118..4140
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA
                     (truncated)"
     primer_bind     4305..4321
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        4331..4349
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    complement(4358..4465)
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     promoter        complement(4478..4496)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4517..4533)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4541..4557)
                     /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(4565..4595)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4610..4631)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     misc_feature    4870..4894
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     misc_feature    4895..4918
                     /note="right border overdrive sequence (Peralta et al.
                     (1986) EMBO J 5: 1137-1142)"
ORIGIN
        1 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
       61 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
      121 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
      181 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
      241 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
      301 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
      361 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
      421 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
      481 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
      541 cttcggaaga agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
      601 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
      661 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
      721 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
      781 atcaatctaa agtatatatg tgtaacattg gtctagtgat tagaaaaact catcgagcat
      841 caaatgaaac tgcaatttat tcatatcagg attatcaata ccatattttt gaaaaagccg
      901 tttctgtaat gaaggagaaa actcaccgag gcagttccat aggatggcaa gatcctggta
      961 tcggtctgcg attccgactc gtccaacatc aatacaacct attaatttcc cctcgtcaaa
     1021 aataaggtta tcaagtgaga aatcaccatg agtgacgact gaatccggtg agaatggcaa
     1081 aagtttatgc atttctttcc agacttgttc aacaggccag ccattacgct cgtcatcaaa
     1141 atcactcgca tcaaccaaac cgttattcat tcgtgattgc gcctgagcga gacgaaatac
     1201 gcgatcgctg ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc gcaggaacac
     1261 tgccagcgca tcaacaatat tttcacctga atcaggatat tcttctaata cctggaatgc
     1321 tgttttccct gggatcgcag tggtgagtaa ccatgcatca tcaggagtac ggataaaatg
     1381 cttgatggtc ggaagaggca taaattccgt cagccagttt agtctgacca tctcatctgt
     1441 aacaacattg gcaacgctac ctttgccatg tttcagaaac aactctggcg catcgggctt
     1501 cccatacaat cggtagattg tcgcacctga ttgcccgaca ttatcgcgag cccatttata
     1561 cccatataaa tcagcatcca tgttggaatt taatcgcggc cttgagcaag acgtttcccg
     1621 ttgaatatgg ctcataacac cccttgtatt actgtttatg taagcagaca gttttattgt
     1681 tcatgatgat atatttttat cttgtgcaat gtaacatcag agattttgag acactcgacg
     1741 gtatcgagct tgatatcgaa ttgatccgaa ttcctttgtt aaccatcatt tagtgacaca
     1801 attaacttaa atttagaaga gactaatttc aacccacaag atggaaatgc aaggacaatg
     1861 ggttcttgaa gttatttgga aacaaagcaa cacaacacaa tagcttattc attagttctc
     1921 atccattaat ttccattgaa gtccgaaaac agaaaccaaa aatacaggat gatgaaatct
     1981 cataaagcaa aaacaaatgg gaatgtagtt taagtacaac aaacaaagtg gggtggctac
     2041 agatgctttg caaatttttc aatgacttac ggatcaattc ctgcagcccc ttaaagctca
     2101 tcatgtttgt atagttcatc catgccatgt gtaatcccag cagctgttac aaactcaaga
     2161 aggaccatgt ggtctctctt ttcgttggga tctttcgaaa gggcagattg tgtggacagg
     2221 taatggttgt ctggtaaaag gacagggcca tcgccaattg gagtattttg ttgataatga
     2281 tcagcgagtt gcacgccgcc gtcttcgatg ttgtggcggg tcttgaagtt ggctttgatg
     2341 ccgttctttt gcttgtcggc catgatgtat acgttgtggg agttgtagtt gtattccaac
     2401 ttgtggccga ggatgtttcc gtcctccttg aaatcgattc ccttaagctc gatcctgttg
     2461 acgagggtgt ctccctcaaa cttgacttca gcacgtgtct tgtagttccc gtcgtccttg
     2521 aagaagatgg tcctctcctg cacgtatccc tcaggcatgg cgctcttgaa gaagtcgtgc
     2581 cgcttcatat gatctgggta tcttgaaaag cattgaacac cataagagaa agtagtgaca
     2641 agtgttggcc atggaacagg tagttttcca gtagtgcaaa taaatttaag ggtaagtttt
     2701 ccgtatgttg catcaccttc accctctcca ctgacagaaa atttgtaccc attaacatca
     2761 ccatctaatt caacaagaat tgggacaact ccagtgaaaa gttcttctcc tttactgaat
     2821 tcggccgagg ataatgatag gagaagtgaa aagatgagaa agagaaaaag attagtcttc
     2881 attgttatat ctccttggat cctctagagt cgaggtcctc tccaaatgaa atgaacttcc
     2941 ttatatagag gaagggtctt gcgaaggata gtgggattgt gcgtcatccc ttacgtcagt
     3001 ggagatatca catcaatcca cttgctttga agacgtggtt ggaacgtctt ctttttccac
     3061 gatgttcctc gtgggtgggg gtccatcttt gggaccactg tcggtagagg catcttgaac
     3121 gatagccttt cctttatcgc aatgatggca tttgtagaag ccatcttcct tttctactgt
     3181 cctttcgatg aagtgacaga tagctgggca atggaatccg aggaggtttc ccgatattac
     3241 cctttgttga aaagtctcaa tagccctctg gtcttctgag actgtatctt tgatattctt
     3301 ggagtagacg agagtgtcgt gctccaccat gttgacctgc aggcatgcaa gctgggggat
     3361 ccactaggtg gctttgttga ataaatcgaa cttttgctga gttgaaggat cagatcacgc
     3421 atcttcccga caacgcagac cgttccgtgg caaagcaaaa gttcaaaatc accaactggt
     3481 ccacctacaa caaagctctc atcaaccgtg gctccctcac tttctggctg gatgatgggg
     3541 cgattcaggc gatccccatc caacagcccg ccgtcgagcg ggctttttta tccccggaag
     3601 cctgtggata gagggtagtt atccacgtga aaccgctaat gccccgcaaa gccttgattc
     3661 acggggcttt ccggcccgct ccaaaaacta tccacgtgaa atcgctaatc agggtacgtg
     3721 aaatcgctaa tcggagtacg tgaaatcgct aataaggtca cgtgaaatcg ctaatcaaaa
     3781 aggcacgtga gaacgctaat agccctttca gatcaacagc ttgcaaacac ccctcgctcc
     3841 ggcaagtagt tacagcaagt agtatgttca attagctttt caattatgaa tatatatatc
     3901 aattattggt cgcccttggc ttgtggacaa tgcgctacgc gcaccggctc cgcccgtgga
     3961 caaccgcaag cggttgccca ccgtcgagcg cctttgccca caacccggcg gccggccgca
     4021 acagatcgtt ttataaattt ttttttttga aaaagaaaaa gcccgaaagg cggcaacctc
     4081 tcgggcttct ggatttccga tccccggaat tagatcttgg caggatatat tgtggtgtaa
     4141 cgttaacatt aacgcttaca atttccattc gccattcagg ctgcgcaact gttgggaagg
     4201 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag
     4261 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag
     4321 tgagcgcgcg taatacgact cactataggg cgaattgggt accgggcccc ccctcgaggt
     4381 cgacggtatc gataagcttg atatcgaatt cctgcagccc gggggatcca ctagttctag
     4441 agcggccgcc accgcggtgg agctccagct tttgttccct ttagtgaggg ttaattgcgc
     4501 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     4561 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     4621 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     4681 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     4741 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     4801 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     4861 tgaaggcctt gacaggatat attggcgggt aaactaagtc gctgtatgtg tttgtttgag
     4921 atct
//
