LOCUS       40924_22588        7065 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Agrobacterium binary vector with kanamycin-resistance and luciferase
            genes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7065)
  AUTHORS   Hellens RP, Edwards EA, Leyland NR, Bean S, Mullineaux PM.
  TITLE     pGreen: a versatile and flexible binary Ti vector for 
            Agrobacterium-mediated plant transformation.
  JOURNAL   Plant Mol. Biol. 2000;42:819-32.
  PUBMED    10890530
REFERENCE   2  (bases 1 to 7065)
  AUTHORS   pGreen website
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 7065)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Plant Mol. 
            Biol."; date: "2000"; volume: "42"; pages: "819-32"
COMMENT     SGRef: number: 2; type: "Journal Article"
COMMENT     This plasmid must be transformed into an Agrobacterium strain that 
            also carries a vector such as pSoup.
FEATURES             Location/Qualifiers
     source          1..7065
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      63..498
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     misc_feature    633..655
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA
                     (truncated)"
     terminator      complement(679..930)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(973..1764)
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERGRTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTQGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     promoter        complement(1798..1977)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     primer_bind     2224..2240
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2247..2265
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    complement(2274..2381)
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     promoter        complement(2394..2412)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2433..2449)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2457..2473)
                     /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(2481..2512)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2527..2548)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     polyA_signal    complement(2787..2963)
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(3082..4731)
                     /codon_start=1
                     /label=luciferase
                     /note="firefly luciferase"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     promoter        complement(4742..5087)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     misc_feature    5148..5172
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(5263..5851)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6025..6837)
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANVVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
ORIGIN
        1 agctctcatc aaccgtggct ccctcacttt ctggctggat gatggggcga ttcaggcgat
       61 ccccatccaa cagcccgccg tcgagcgggc ttttttatcc ccggaagcct gtggatagag
      121 ggtagttatc cacgtgaaac cgctaatgcc ccgcaaagcc ttgattcacg gggctttccg
      181 gcccgctcca aaaactatcc acgtgaaatc gctaatcagg gtacgtgaaa tcgctaatcg
      241 gagtacgtga aatcgctaat aaggtcacgt gaaatcgcta atcaaaaagg cacgtgagaa
      301 cgctaatagc cctttcagat caacagcttg caaacacccc tcgctccggc aagtagttac
      361 agcaagtagt atgttcaatt agcttttcaa ttatgaatat atatatcaat tattggtcgc
      421 ccttggcttg tggacaatgc gctacgcgca ccggctccgc ccgtggacaa ccgcaagcgg
      481 ttgcccaccg tcgagcgcca gcgcctttgc ccacaacccg gcggccggcc gcaacagatc
      541 gttttataaa tttttttttt tgaaaaagaa aaagcccgaa aggcggcaac ctctcgggct
      601 tctggatttc cgatccccgg aattagagat cttggcagga tatattgtgg tgtaacgtta
      661 tcagcttgca tgccggtcga tctagtaaca tagatgacac cgcgcgcgat aatttatcct
      721 agtttgcgcg ctatattttg ttttctatcg cgtattaaat gtataattgc gggactctaa
      781 tcaaaaaacc catctcataa ataacgtcat gcattacatg ttaattatta catgcttaac
      841 gtaattcaac agaaattata tgataatcat cgcaagaccg gcaacaggat tcaatcttaa
      901 gaaactttat tgccaaatgt ttgaacgatc tgcttgactc tagctagagt ccgaacccca
      961 gagtcccgct cagaagaact cgtcaagaag gcgatagaag gcgatgcgct gcgaatcggg
     1021 agcggcgata ccgtaaagca cgaggaagcg gtcagcccat tcgccgccaa gctcttcagc
     1081 aatatcacgg gtagccaacg ctatgtcctg atagcggtcc gccacaccca gccggccaca
     1141 gtcgatgaat ccagaaaagc ggccattttc caccatgata ttcggcaagc aggcatcgcc
     1201 ctgggtcacg acgagatcct cgccgtcggg catccgcgcc ttgagcctgg cgaacagttc
     1261 ggctggcgcg agcccctgat gctcttcgtc cagatcatcc tgatcgacaa gaccggcttc
     1321 catccgagta cgtcctcgct cgatgcgatg tttcgcttgg tggtcgaatg ggcaggtagc
     1381 cggatcaagc gtatgcagcc gccgcattgc atcagccatg atggatactt tctcggcagg
     1441 agcaaggtga gatgacagga gatcctgccc cggcacttcg cccaatagca gccagtccct
     1501 tcccgcttca gtgacaacgt cgagcacagc tgcgcaagga acgcccgtcg tggccagcca
     1561 cgatagccgc gctgcctcgt cttggagttc attcagggca ccggacaggt cggtcttgac
     1621 aaaaagaacc gggcgcccct gcgctgacag ccggaacacg gcggcatcag agcagccgat
     1681 tgtctgttgt gcccagtcat agccgaatag cctctccacc caagcggccg gagaacctgc
     1741 gtgcaatcca tcttgttcaa tcatgcctcg atcgagttga gagtgaatat gagactctaa
     1801 ttggataccg aggggaattt atggaacgtc agtggagcat ttttgacaag aaatatttgc
     1861 tagctgatag tgaccttagg cgacttttga acgcgcaata atggtttctg acgtatgtgc
     1921 ttagctcatt aaactccaga aacccgcggc tgagtggctc cttcaacgtt gcggttctgt
     1981 cagttccaaa cgtaaaacgg cttgtcccgc gtcatcggcg ggggtcataa cgtgactccc
     2041 ttaattctca tgtatcgata acattaacgt ttacaatttc gcgccattcg ccattcaggc
     2101 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga
     2161 aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac
     2221 gttgtaaaac gacggccagt gaattgtaat acgactcact atagggcgaa ttgggtaccg
     2281 ggccccccct cgaggtcgac ggtatcgata agcttgatat cgaattcctg cagcccgggg
     2341 gatccactag ttctagagcg gccgccaccg cggtggagct ccagcttttg ttccctttag
     2401 tgagggttaa ttccgagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     2461 tatccgctca caattccaca caacatacga gccggaaghc ataaagtgta aagcctgggg
     2521 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     2581 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     2641 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     2701 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     2761 taacgcagga aagaacatga aggatcgatc tggattttag tactggattt tggttttagg
     2821 aattagaaat tttattgata gaagtatttt acaaatacaa atacatacta agggtttctt
     2881 atatgctcaa cacatgagcg aaaccctata agaaccctaa tttcccttat cgggaaacta
     2941 ctcacacatt atttatggag aaaatagaga gagatagatt tgtagagaga gactggtgat
     3001 ttcagcgtac cgaattaatt ctctagctag agtattacaa tagctaagaa tttcgtcatc
     3061 gctgaataca gttacatttt acaatttgga ctttccgccc ttcttggcct ttatgaggat
     3121 ctctctgatt tttcttgcgt cgagttttcc ggtaagacct ttcggtactt cgtccacaaa
     3181 cacaactcct ccgcgcaact ttttggcggt tgttacttga ctggcgacgt aatccacgat
     3241 ctctttttcc gtcatcgtct ttccgtgctc caaaacaaca acggcggcgg gaagttcacc
     3301 ggcgtcatcg tcgggaagac ctgccacgcc cgcgtcgaag atgttggggt gttgtaacaa
     3361 tatcgactcc aattcagcgg gggccacctg gtatcctttg tatttaatta aagacttcaa
     3421 gcggtcaact atgaagaagt gttcgtcttc gtcccagtaa gctatgtctc cagaatgtag
     3481 ccatccatcc ttgtcaatca aggcgttggt cgcttccgga ttgtttacat aaccggacat
     3541 aatcataggt cctctgacac ataattcgcc tctctgatta acgcccagcg ttttcccggt
     3601 atccagatcc acaaccttcg cttcaaaaaa tggaacaact ttaccgaccg cgcccggttt
     3661 atcatccccc tcgggtgtaa tcagaatagc tgatgtagtc tcagtgagcc catatccttg
     3721 tcgtatccct ggaagatgga agcgttttgc aaccgcttcc ccgacttctt tcgaaagagg
     3781 tgcgccccca gaagcaattt cgtgtaaatt agataaatcg tatttgtcaa tcagagtgct
     3841 tttggcgaag aatgaaaata gggttggtac tagcaacgca ctttgaattt tgtaatcctg
     3901 aagggatcgt aaaaacagct cttcttcaaa tctatacatt aagacgactc gaaatccaca
     3961 tatcaaatat ccgagtgtag taaacattcc aaaaccgtga tggaatggaa caacacttaa
     4021 aatcgcagta tccggaatga tttgattgcc aaaaatagga tctctggcat gcgagaatct
     4081 gacgcaggca gttctatgcg gaagggccac acccttaggt aacccagtag atccagagga
     4141 gttcattatc agtgcaattg ttttgtcacg atcaaaggac tctggtacaa aatcgtattc
     4201 attaaaaccg ggaggtagat gagatgtgac gaacgtgtac atcgactgaa atccctggta
     4261 atccgtttta gaatccatga taataatttt ctggattatt ggtaattttt tttgcacgtt
     4321 caaaattttt tgcaacccct ttttggaaac aaacactacg gtaggctgcg aaatgttcat
     4381 actgttgagc aattcacgtt cattataaat gtcgttcgcg ggcgcaactg caactccgat
     4441 aaataacgcg cccaacaccg gcataaagaa ttgaagagag ttttcactgc atacgacgat
     4501 tctgtgattt gtattcagcc catatcgttt catagcttct gccaaccgaa cggacatttc
     4561 gaagtattcc gcgtacgtga tgttcacctc gatatgtgca tctgtaaaag caattgttcc
     4621 aggaaccagg gcgtatctct tcatagcctt atgcagttgc tctccagcgg ttccatcctc
     4681 tagcggatag aatggcgccg ggcctttctt tatgtttttg gcgtcttcca tgagcttggg
     4741 ctgtcctctc caaatgaaat gaacttcctt atatagagga agggtcttgc gaaggatagt
     4801 gggattgtgc gtcatccctt acgtcagtgg agatgtcaca tcaatccact tgctttgaag
     4861 acgtggttgg aacgtcttct ttttccacga tgctcctcgt gggtgggggt ccatctttgg
     4921 gaccactgtc ggcagaggca tcttgaatga tagcctttcc tttatcgcaa tgatggcatt
     4981 tgtaggagcc accttccttt tctactgtcc tttcgatgaa gtgacagata gctgggcaat
     5041 ggaatccgag gaggtttccc gaaattaccc tttgttgaaa agtctcaata gccctttggt
     5101 cttctgagac tgtatctttg acatttttgg agtaggggta cgatccttga caggatatat
     5161 tggcgggtaa actaagtcgc tgtatgtgtt tgtttgagat ctcatgtgag caaaaggcca
     5221 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     5281 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     5341 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     5401 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     5461 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     5521 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     5581 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     5641 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     5701 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa gaagagttgg
     5761 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     5821 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     5881 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     5941 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     6001 tgtgtaacat tggtctagtg attagaaaaa ctcatcgagc atcaaatgaa actgcaattt
     6061 attcatatca ggattatcaa taccatattt ttgaaaaagc cgtttctgta atgaaggaga
     6121 aaactcaccg aggcagttcc ataggatggc aagatcctgg tatcggtctg cgattccgac
     6181 tcgtccaaca tcaatacaac ctattaattt cccctcgtca aaaataaggt tatcaagtga
     6241 gaaatcacca tgagtgacga ctgaatccgg tgagaatggc aaaagtttat gcatttcttt
     6301 ccagacttgt tcaacaggcc agccattacg ctcgtcatca aaatcactcg catcaaccaa
     6361 accgttattc attcgtgatt gcgcctgagc gagacgaaat acgcgatcgc tgttaaaagg
     6421 acaattacaa acaggaatcg aatgcaaccg gcgcaggaac actgccagcg catcaacaat
     6481 attttcacct gaatcaggat attcttctaa tacctggaat gctgttttcc ctgggatcgc
     6541 agtggtgagt aaccatgcat catcaggagt acggataaaa tgcttgatgg tcggaagagg
     6601 cataaattcc gtcagccagt ttagtctgac catctcatct gtaacaacat tggcaacgct
     6661 acctttgcca tgtttcagaa acaactctgg cgcatcgggc ttcccataca atcggtagat
     6721 tgtcgcacct gattgcccga cattatcgcg agcccattta tacccatata aatcagcatc
     6781 catgttggaa tttaatcgcg gccttgagca agacgtttcc cgttgaatat ggctcataac
     6841 accccttgta ttactgttta tgtaagcaga cagttttatt gttcatgatg atatattttt
     6901 atcttgtgca atgtaacatc agagattttg agacacaacg tggctttgtt gaataaatcg
     6961 aacttttgct gagttgaagg atcagatcac gcatcttccc gacaacgcag accgttccgt
     7021 ggcaaagcaa aagttcaaaa tcaccaactg gtccacctac aacaa
//
