LOCUS       40924_10996        4971 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Binary vector pCLEAN-G146, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4971)
  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 4971)
  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 4971)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4971)
  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..4971
                     /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"
     rep_origin      1926..2361
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     regulatory      complement(2538..2834)
                     /label=soybean-derived poly(A)signal
                     /note="soybean-derived poly(A)signal"
                     /regulatory_class="terminator"
     CDS             complement(2870..3580)
                     /label=mgfp5
                     /note="GFP with folding enhancement mutations"
     promoter        complement(3679..4023)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     misc_feature    4141..4165
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     primer_bind     4338..4354
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        4364..4382
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    complement(4391..4498)
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     promoter        complement(4511..4529)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4550..4566)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4574..4590)
                     /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(4598..4628)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4643..4664)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     misc_feature    4911..4935
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     misc_feature    4936..4949
                     /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 acacaacgtg
     1741 gctttgttga ataaatcgaa cttttgctga gttgaaggat cagatcacgc atcttcccga
     1801 caacgcagac cgttccgtgg caaagcaaaa gttcaaaatc accaactggt ccacctacaa
     1861 caaagctctc atcaaccgtg gctccctcac tttctggctg gatgatgggg cgattcaggc
     1921 gatccccatc caacagcccg ccgtcgagcg ggctttttta tccccggaag cctgtggata
     1981 gagggtagtt atccacgtga aaccgctaat gccccgcaaa gccttgattc acggggcttt
     2041 ccggcccgct ccaaaaacta tccacgtgaa atcgctaatc agggtacgtg aaatcgctaa
     2101 tcggagtacg tgaaatcgct aataaggtca cgtgaaatcg ctaatcaaaa aggcacgtga
     2161 gaacgctaat agccctttca gatcaacagc ttgcaaacac ccctcgctcc ggcaagtagt
     2221 tacagcaagt agtatgttca attagctttt caattatgaa tatatatatc aattattggt
     2281 cgcccttggc ttgtggacaa tgcgctacgc gcaccggctc cgcccgtgga caaccgcaag
     2341 cggttgccca ccgtcgagcg cctttgccca caacccggcg gccggccgca acagatcgtt
     2401 ttataaattt ttttttttga aaaagaaaaa gcccgaaagg cggcaacctc tcgggcttct
     2461 ggatttccga tccccggaat tagatccgtt taaactactc gacggtatcg agcttgatat
     2521 cgaattgatc cgaattcctt tgttaaccat catttagtga cacaattaac ttaaatttag
     2581 aagagactaa tttcaaccca caagatggaa atgcaaggac aatgggttct tgaagttatt
     2641 tggaaacaaa gcaacacaac acaatagctt attcattagt tctcatccat taatttccat
     2701 tgaagtccga aaacagaaac caaaaataca ggatgatgaa atctcataaa gcaaaaacaa
     2761 atgggaatgt agtttaagta caacaaacaa agtggggtgg ctacagatgc tttgcaaatt
     2821 tttcaatgac ttacggatca attcctgcag ccccttaaag ctcatcatgt ttgtatagtt
     2881 catccatgcc atgtgtaatc ccagcagctg ttacaaactc aagaaggacc atgtggtctc
     2941 tcttttcgtt gggatctttc gaaagggcag attgtgtgga caggtaatgg ttgtctggta
     3001 aaaggacagg gccatcgcca attggagtat tttgttgata atgatcagcg agttgcacgc
     3061 cgccgtcttc gatgttgtgg cgggtcttga agttggcttt gatgccgttc ttttgcttgt
     3121 cggccatgat gtatacgttg tgggagttgt agttgtattc caacttgtgg ccgaggatgt
     3181 ttccgtcctc cttgaaatcg attcccttaa gctcgatcct gttgacgagg gtgtctccct
     3241 caaacttgac ttcagcacgt gtcttgtagt tcccgtcgtc cttgaagaag atggtcctct
     3301 cctgcacgta tccctcaggc atggcgctct tgaagaagtc gtgccgcttc atatgatctg
     3361 ggtatcttga aaagcattga acaccataag agaaagtagt gacaagtgtt ggccatggaa
     3421 caggtagttt tccagtagtg caaataaatt taagggtaag ttttccgtat gttgcatcac
     3481 cttcaccctc tccactgaca gaaaatttgt acccattaac atcaccatct aattcaacaa
     3541 gaattgggac aactccagtg aaaagttctt ctcctttact gaattcggcc gaggataatg
     3601 ataggagaag tgaaaagatg agaaagagaa aaagattagt cttcattgtt atatctcctt
     3661 ggatcctcta gagtcgaggt cctctccaaa tgaaatgaac ttccttatat agaggaaggg
     3721 tcttgcgaag gatagtggga ttgtgcgtca tcccttacgt cagtggagat atcacatcaa
     3781 tccacttgct ttgaagacgt ggttggaacg tcttcttttt ccacgatgtt cctcgtgggt
     3841 gggggtccat ctttgggacc actgtcggta gaggcatctt gaacgatagc ctttccttta
     3901 tcgcaatgat ggcatttgta gaagccatct tccttttcta ctgtcctttc gatgaagtga
     3961 cagatagctg ggcaatggaa tccgaggagg tttcccgata ttaccctttg ttgaaaagtc
     4021 tcaatagccc tctggtcttc tgagactgta tctttgatat tcttggagta gacgagagtg
     4081 tcgtgctcca ccatgttgac ctgcaggcat gcaagctggg ggatccacta ggtaagatct
     4141 tggcaggata tattgtggtg taaacgttcc tgcggccaac attaacgctt acaatttcca
     4201 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt
     4261 acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt
     4321 ttcccagtca cgacgttgta aaacgacggc cagtgagcgc gcgtaatacg actcactata
     4381 gggcgaattg ggtaccgggc cccccctcga ggtcgacggt atcgataagc ttgatatcga
     4441 attcctgcag cccgggggat ccactagttc tagagcggcc gccaccgcgg tggagctcca
     4501 gcttttgttc cctttagtga gggttaattg cgcgcttggc gtaatcatgg tcatagctgt
     4561 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     4621 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     4681 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     4741 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     4801 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     4861 ccacagaatc aggggataac gcaggaaaga acatgaaggt cgacgttcct tgacaggata
     4921 tattggcggg taaactaagt cgctgtatgt gtttgtttga gatctggatc t
//
