LOCUS       40924_8821        8590 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Binary vector pCAMBIA-EGFP, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8590)
  AUTHORS   Dalal J, Yalamanchili R, La Hovary C, Ji M, Rodriguez-Welsh M, 
            Aslett D, Ganapathy S, Grunden A, Sederoff H, Qu R.
  TITLE     A novel gateway-compatible binary vector series (PC-GW) for flexible
            cloning of multiple genes for genetic transformation of plants
  JOURNAL   Plasmid 81, 55-62 (2015)
  PUBMED    26188330
REFERENCE   2  (bases 1 to 8590)
  AUTHORS   Dalal J, Yalamanchili R, Hovary CL, Ji M, Rodriguez-Welsh M, Aslett 
            D, Ganapathy S, Grunden A, Sederoff H, Qu R.
  TITLE     Direct Submission
  JOURNAL   Submitted (18-FEB-2015) Crop Science, North Carolina State 
            University, Campus Box 7287, Raleigh, NC 27695, USA
REFERENCE   3  (bases 1 to 8590)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8590)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Plasmid 81,
            55-62 (2015)"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (18-FEB-2015) Crop Science, North Carolina State University, Campus 
            Box 7287, Raleigh, NC 27695, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8590
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1219..1845
                     /codon_start=1
                     /label=pVS1 StaA
                     /note="stability protein from the Pseudomonas plasmid pVS1
                     (Heeb et al., 2000)"
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             2282..3346
                     /codon_start=1
                     /label=pVS1 RepA
                     /note="replication protein from the Pseudomonas plasmid
                     pVS1 (Heeb et al., 2000)"
                     /translation="GRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESWQA
                     AADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLSKR
                     DRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKPGR
                     VFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGEAL
                     ISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYRLA
                     RRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPILV
                     MRYRNLIEGEASAGS"
     rep_origin      3415..3609
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid
                     pVS1 (Heeb et al., 2000)"
     misc_feature    3953..4093
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(4279..4867)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4957..5748)
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     misc_feature    6173..6197
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     polyA_signal    complement(6275..6449)
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(6459..7175)
                     /codon_start=1
                     /label=EGFP
                     /note="enhanced GFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     promoter        complement(7214..7891)
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a
                     duplicated enhancer region"
     protein_bind    8082..8103
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        8118..8148
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8156..8172
                     /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)."
     primer_bind     8180..8196
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    8206..8262
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(8266..8282)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    8485..8509
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
ORIGIN
        1 catgccaacc acagggttcc cctcgggatc aaagtacttt gatccaaccc ctccgctgct
       61 atagtgcagt cggcttctga cgttcagtgc agccgtcttc tgaaaacgac atgtcgcaca
      121 agtcctaagt tacgcgacag gctgccgccc tgcccttttc ctggcgtttt cttgtcgcgt
      181 gttttagtcg cataaagtag aatacttgcg actagaaccg gagacattac gccatgaaca
      241 agagcgccgc cgctggcctg ctgggctatg cccgcgtcag caccgacgac caggacttga
      301 ccaaccaacg ggccgaactg cacgcggccg gctgcaccaa gctgttttcc gagaagatca
      361 ccggcaccag gcgcgaccgc ccggagctgg ccaggatgct tgaccaccta cgccctggcg
      421 acgttgtgac agtgaccagg ctagaccgcc tggcccgcag cacccgcgac ctactggaca
      481 ttgccgagcg catccaggag gccggcgcgg gcctgcgtag cctggcagag ccgtgggccg
      541 acaccaccac gccggccggc cgcatggtgt tgaccgtgtt cgccggcatt gccgagttcg
      601 agcgttccct aatcatcgac cgcacccgga gcgggcgcga ggccgccaag gcccgaggcg
      661 tgaagtttgg cccccgccct accctcaccc cggcacagat cgcgcacgcc cgcgagctga
      721 tcgaccagga aggccgcacc gtgaaagagg cggctgcact gcttggcgtg catcgctcga
      781 ccctgtaccg cgcacttgag cgcagcgagg aagtgacgcc caccgaggcc aggcggcgcg
      841 gtgccttccg tgaggacgca ttgaccgagg ccgacgccct ggcggccgcc gagaatgaac
      901 gccaagagga acaagcatga aaccgcacca ggacggccag gacgaaccgt ttttcattac
      961 cgaagagatc gaggcggaga tgatcgcggc cgggtacgtg ttcgagccgc ccgcgcacgt
     1021 ctcaaccgtg cggctgcatg aaatcctggc cggtttgtct gatgccaagc tggcggcctg
     1081 gccggccagc ttggccgctg aagaaaccga gcgccgccgt ctaaaaaggt gatgtgtatt
     1141 tgagtaaaac agcttgcgtc atgcggtcgc tgcgtatatg atgcgatgag taaataaaca
     1201 aatacgcaag gggaacgcat gaaggttatc gctgtactta accagaaagg cgggtcaggc
     1261 aagacgacca tcgcaaccca tctagcccgc gccctgcaac tcgccggggc cgatgttctg
     1321 ttagtcgatt ccgatcccca gggcagtgcc cgcgattggg cggccgtgcg ggaagatcaa
     1381 ccgctaaccg ttgtcggcat cgaccgcccg acgattgacc gcgacgtgaa ggccatcggc
     1441 cggcgcgact tcgtagtgat cgacggagcg ccccaggcgg cggacttggc tgtgtccgcg
     1501 atcaaggcag ccgacttcgt gctgattccg gtgcagccaa gcccttacga catatgggcc
     1561 accgccgacc tggtggagct ggttaagcag cgcattgagg tcacggatgg aaggctacaa
     1621 gcggcctttg tcgtgtcgcg ggcgatcaaa ggcacgcgca tcggcggtga ggttgccgag
     1681 gcgctggccg ggtacgagct gcccattctt gagtcccgta tcacgcagcg cgtgagctac
     1741 ccaggcactg ccgccgccgg cacaaccgtt cttgaatcag aacccgaggg cgacgctgcc
     1801 cgcgaggtcc aggcgctggc cgctgaaatt aaatcaaaac tcatttgagt taatgaggta
     1861 aagagaaaat gagcaaaagc acaaacacgc taagtgccgg ccgtccgagc gcacgcagca
     1921 gcaaggctgc aacgttggcc agcctggcag acacgccagc catgaagcgg gtcaactttc
     1981 agttgccggc ggaggatcac accaagctga agatgtacgc ggtacgccaa ggcaagacca
     2041 ttaccgagct gctatctgaa tacatcgcgc agctaccaga gtaaatgagc aaatgaataa
     2101 atgagtagat gaattttagc ggctaaagga ggcggcatgg aaaatcaaga acaaccaggc
     2161 accgacgccg tggaatgccc catgtgtgga ggaacgggcg gttggccagg cgtaagcggc
     2221 tgggttgtct gccggccctg caatggcact ggaaccccca agcccgagga atcggcgtga
     2281 cggtcgcaaa ccatccggcc cggtacaaat cggcgcggcg ctgggtgatg acctggtgga
     2341 gaagttgaag gccgcgcagg ccgcccagcg gcaacgcatc gaggcagaag cacgccccgg
     2401 tgaatcgtgg caagcggccg ctgatcgaat ccgcaaagaa tcccggcaac cgccggcagc
     2461 cggtgcgccg tcgattagga agccgcccaa gggcgacgag caaccagatt ttttcgttcc
     2521 gatgctctat gacgtgggca cccgcgatag tcgcagcatc atggacgtgg ccgttttccg
     2581 tctgtcgaag cgtgaccgac gagctggcga ggtgatccgc tacgagcttc cagacgggca
     2641 cgtagaggtt tccgcagggc cggccggcat ggccagtgtg tgggattacg acctggtact
     2701 gatggcggtt tcccatctaa ccgaatccat gaaccgatac cgggaaggga agggagacaa
     2761 gcccggccgc gtgttccgtc cacacgttgc ggacgtactc aagttctgcc ggcgagccga
     2821 tggcggaaag cagaaagacg acctggtaga aacctgcatt cggttaaaca ccacgcacgt
     2881 tgccatgcag cgtacgaaga aggccaagaa cggccgcctg gtgacggtat ccgagggtga
     2941 agccttgatt agccgctaca agatcgtaaa gagcgaaacc gggcggccgg agtacatcga
     3001 gatcgagcta gctgattgga tgtaccgcga gatcacagaa ggcaagaacc cggacgtgct
     3061 gacggttcac cccgattact ttttgatcga tcccggcatc ggccgttttc tctaccgcct
     3121 ggcacgccgc gccgcaggca aggcagaagc cagatggttg ttcaagacga tctacgaacg
     3181 cagtggcagc gccggagagt tcaagaagtt ctgtttcacc gtgcgcaagc tgatcgggtc
     3241 aaatgacctg ccggagtacg atttgaagga ggaggcgggg caggctggcc cgatcctagt
     3301 catgcgctac cgcaacctga tcgagggcga agcatccgcc ggttcctaat gtacggagca
     3361 gatgctaggg caaattgccc tagcagggga aaaaggtcga aaaggtctct ttcctgtgga
     3421 tagcacgtac attgggaacc caaagccgta cattgggaac cggaacccgt acattgggaa
     3481 cccaaagccg tacattggga accggtcaca catgtaagtg actgatataa aagagaaaaa
     3541 aggcgatttt tccgcctaaa actctttaaa acttattaaa actcttaaaa cccgcctggc
     3601 ctgtgcataa ctgtctggcc agcgcacagc cgaagagctg caaaaagcgc ctacccttcg
     3661 gtcgctgcgc tccctacgcc ccgccgcttc gcgtcggcct atcgcggccg ctggccgctc
     3721 aaaaatggct ggcctacggc caggcaatct accagggcgc ggacaagccg cgccgtcgcc
     3781 actcgaccgc cggcgcccac atcaaggcac cctgcctcgc gcgtttcggt gatgacggtg
     3841 aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg
     3901 ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggcgcagcca
     3961 tgacccagtc acgtagcgat agcggagtgt atactggctt aactatgcgg catcagagca
     4021 gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa
     4081 ataccgcatc aggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     4141 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     4201 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     4261 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     4321 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     4381 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     4441 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     4501 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     4561 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     4621 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     4681 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     4741 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     4801 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     4861 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     4921 acgttaaggg attttggtca tgcattctag gtactaaaac aattcatcca gtaaaatata
     4981 atattttatt ttctcccaat caggcttgat ccccagtaag tcaaaaaata gctcgacata
     5041 ctgttcttcc ccgatatcct ccctgatcga ccggacgcag aaggcaatgt cataccactt
     5101 gtccgccctg ccgcttctcc caagatcaat aaagccactt actttgccat ctttcacaaa
     5161 gatgttgctg tctcccaggt cgccgtggga aaagacaagt tcctcttcgg gcttttccgt
     5221 ctttaaaaaa tcatacagct cgcgcggatc tttaaatgga gtgtcttctt cccagttttc
     5281 gcaatccaca tcggccagat cgttattcag taagtaatcc aattcggcta agcggctgtc
     5341 taagctattc gtatagggac aatccgatat gtcgatggag tgaaagagcc tgatgcactc
     5401 cgcatacagc tcgataatct tttcagggct ttgttcatct tcatactctt ccgagcaaag
     5461 gacgccatcg gcctcactca tgagcagatt gctccagcca tcatgccgtt caaagtgcag
     5521 gacctttgga acaggcagct ttccttccag ccatagcatc atgtcctttt cccgttccac
     5581 atcataggtg gtccctttat accggctgtc cgtcattttt aaatataggt tttcattttc
     5641 tcccaccagc ttatatacct tagcaggaga cattccttcc gtatctttta cgcagcggta
     5701 tttttcgatc agttttttca attccggtga tattctcatt ttagccattt attatttcct
     5761 tcctcttttc tacagtattt aaagataccc caagaagcta attataacaa gacgaactcc
     5821 aattcactgt tccttgcatt ctaaaacctt aaataccaga aaacagcttt ttcaaagttg
     5881 ttttcaaagt tggcgtataa catagtatcg acggagccga ttttgaaacc gcggtgatca
     5941 caggcagcaa cgctctgtca tcgttacaat caacatgcta ccctccgcga gatcatccgt
     6001 gtttcaaacc cggcagctta gttgccgttc ttccgaatag catcggtaac atgagcaaag
     6061 tctgccgcct tacaacggct ctcccgctga cgccgtcccg gactgatggg ctgcctgtat
     6121 cgagtggtga ttttgtgccg agctgccggt cggggagctg ttggctggct ggtggcagga
     6181 tatattgtgg tgtaaacaaa ttgacgctta gacaacttaa taacacattg cggacgtttt
     6241 taatgtactg aattaacgcc gaattaattc gggggatctg gattttagta ctggattttg
     6301 gttttaggaa ttagaaattt tattgataga agtattttac aaatacaaat acatactaag
     6361 ggtttcttat atgctcaaca catgagcgaa accctatagg aaccctaatt cccttatctg
     6421 ggaactactc acacattatt atggagaaac tcgagttact tgtacagctc gtccatgccg
     6481 agagtgatcc cggcggcggt cacgaactcc agcaggacca tgtgatcgcg cttctcgttg
     6541 gggtctttgc tcagggcgga ctgggtgctc aggtagtggt tgtcgggcag cagcacgggg
     6601 ccgtcgccga tgggggtgtt ctgctggtag tggtcggcga gctgcacgct gccgtcctcg
     6661 atgttgtggc ggatcttgaa gttcaccttg atgccgttct tctgcttgtc ggccatgata
     6721 tagacgttgt ggctgttgta gttgtactcc agcttgtgcc ccaggatgtt gccgtcctcc
     6781 ttgaagtcga tgcccttcag ctcgatgcgg ttcaccaggg tgtcgccctc gaacttcacc
     6841 tcggcgcggg tcttgtagtt gccgtcgtcc ttgaagaaga tggtgcgctc ctggacgtag
     6901 ccttcgggca tggcggactt gaagaagtcg tgctgcttca tgtggtcggg gtagcggctg
     6961 aagcactgca cgccgtaggt cagggtggtc acgagggtgg gccagggcac gggcagcttg
     7021 ccggtggtgc agatgaactt cagggtcagc ttgccgtagg tggcatcgcc ctcgccctcg
     7081 ccggacacgc tgaacttgtg gccgtttacg tcgccgtcca gctcgaccag gatgggcacc
     7141 accccggtga acagctcctc gcccttgctc accatctcga gagagataga tttgtagaga
     7201 gagactggtg atttcagcgt gtcctctcca aatgaaatga acttccttat atagaggaag
     7261 gtcttgcgaa ggatagtggg attgtgcgtc atcccttacg tcagtggaga tatcacatca
     7321 atccacttgc tttgaagacg tggttggaac gtcttctttt tccacgatgc tcctcgtggg
     7381 tgggggtcca tctttgggac cactgtcggc agaggcatct tgaacgatag cctttccttt
     7441 atcgcaatga tggcatttgt aggtgccacc ttccttttct actgtccttt tgatgaagtg
     7501 acagatagct gggcaatgga atccgaggag gtttcccgat attacccttt gttgaaaagt
     7561 ctcaatagcc ctttggtctt ctgagactgt atctttgata ttcttggagt agacgagagt
     7621 gtcgtgctcc accatgttat cacatcaatc cacttgcttt gaagacgtgg ttggaacgtc
     7681 ttctttttcc acgatgctcc tcgtgggtgg gggtccatct ttgggaccac tgtcggcaga
     7741 ggcatcttga acgatagcct ttcctttatc gcaatgatgg catttgtagg tgccaccttc
     7801 cttttctact gtccttttga tgaagtgaca gatagctggg caatggaatc cgaggaggtt
     7861 tcccgatatt accctttgtt gaaaagtctc aatagccctt tggtcttctg agactgtatc
     7921 tttgatattc ttggagtaga cgagagtgtc gtgctccacc atgttggcaa gctgctctag
     7981 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     8041 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
     8101 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
     8161 agcggataac aatttcacac aggaaacagc tatgaccatg attacgaatt cgagctcggt
     8221 acccggggat cctctagagt cgacctgcag gcatgcaagc ttggcactgg ccgtcgtttt
     8281 acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc
     8341 ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
     8401 gcgcagcctg aatggcgaat gctagagcag cttgagcttg gatcagattg tcgtttcccg
     8461 ccttcagttt aaactatcag tgtttgacag gatatattgg cgggtaaacc taagagaaaa
     8521 gagcgtttat tagaataacg gatatttaaa agggcgtgaa aaggtttatc cgttcgtcca
     8581 tttgtatgtg
//
