LOCUS       pVP13.        7675 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Bacterial Gateway(R) destination vector for appending an N-terminal 
            S-Tag-6xHis-MBP cassette to an expressed protein. Also known as 
            pVP13-GW.
ACCESSION   .
VERSION     .
KEYWORDS    pVP13.
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7675)
  AUTHORS   Thao S, Zhao Q, Kimball T, Steffen E, Blommel PG, Riters M, Newman 
            CS, Fox BG, Wrobel RL.
  TITLE     Results from high-throughput DNA cloning of Arabidopsis thaliana 
            target genes using site-specific recombination.
  JOURNAL   J. Struct. Funct. Genomics 2004;5:267-76.
  PUBMED    15750721
REFERENCE   2  (bases 1 to 7675)
  AUTHORS   Center for Eukaryotic Structural Genomics
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 7675)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J. Struct. 
            Funct. Genomics"; date: "2004"; volume: "5"; pages: "267-76"
COMMENT     SGRef: number: 2; type: "Journal Article"
COMMENT     A TEV cleavage site can be encoded during PCR amplification.
FEATURES             Location/Qualifiers
     source          1..7675
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        11..55
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA
                     polymerase, with embedded lac operator"
     protein_bind    63..79
                     /label=lac repressor encoded by lacI binding site
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="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)."
     RBS             102..107
                     /note="ribosome binding site"
     CDS             116..118
                     /codon_start=1
                     /product="start codon"
                     /label=start codon
                     /note="ATG"
                     /translation="M"
     CDS             122..166
                     /label=S-Tag
                     /note="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
     CDS             173..190
                     /label=6xHis
                     /note="6xHis affinity tag"
     CDS             194..1291
                     /label=MBP
                     /note="maltose binding protein from E. coli"
     protein_bind    1361..1485
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        1510..1540
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             1594..2250
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     CDS             2595..2897
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     protein_bind    complement(2941..3065)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     terminator      3133..3227
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             3271..3927
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     terminator      3995..4081
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     protein_bind    complement(4140..4161)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(4177..5256)
                     /label=lacI
                     /note="lac repressor"
     promoter        complement(5257..5334)
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the
                     promoter boosts expression of the lacI gene about 10-fold."
     misc_feature    5526..5666
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(5852..6440)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6614..7471)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(7472..7576)
                     /label=AmpR promoter
ORIGIN
        1 cctcgagaaa tcataaaaaa tttatttgct ttgtgagcgg ataacaatta taatagattc
       61 aattgtgagc ggataacaat ttcacacaga attcattaaa gaggagaaat taaccatggg
      121 taaagaaacc gctgctgcga aatttgaacg ccagcacatg gactcgccac cgcatcatca
      181 ccatcaccat tcaaaaatcg aagaaggtaa actggtaatc tggattaacg gcgataaagg
      241 ctataacggt ctcgctgaag tcggtaagaa attcgagaaa gataccggaa ttaaagtcac
      301 cgttgagcat ccggataaac tggaagagaa attcccacag gttgcggcaa ctggcgatgg
      361 ccctgacatt atcttctggg cacacgaccg ctttggtggc tacgctcaat ctggcctgtt
      421 ggctgaaatc accccggaca aagcgttcca ggacaagctg tatccgttta cctgggatgc
      481 cgtacgttac aacggcaagc tgattgctta cccgatcgct gttgaagcgt tatcgctgat
      541 ttataacaaa gatctgctgc cgaacccgcc aaaaacctgg gaagagatcc cggcgctgga
      601 taaagaactg aaagcgaaag gtaagagcgc gctgatgttc aacctgcaag aaccgtactt
      661 cacctggccg ctgattgctg ctgacggggg ttatgcgttc aagtatgaaa acggcaagta
      721 cgacattaaa gacgtgggcg tggataacgc tggcgcgaaa gcgggtctga ccttcctggt
      781 tgacctgatt aaaaacaaac acatgaatgc agacaccgat tactccatcg cagaagctgc
      841 ctttaataaa ggcgaaacag cgatgaccat caacggcccg tgggcatggt ccaacatcga
      901 caccagcaaa gtgaattatg gtgtaacggt actgccgacc ttcaagggtc aaccatccaa
      961 accgttcgtt ggcgtgctga gcgcaggtat taacgccgcc agtccgaaca aagagctggc
     1021 aaaagagttc ctcgaaaact atctgctgac tgatgaaggt ctggaagcgg ttaataaaga
     1081 caaaccgctg ggtgccgtag cgctgaagtc ttacgaggaa gagttggcga aagatccacg
     1141 tattgccgcc actatggaaa acgcccagaa aggtgaaatc atgccgaaca tcccgcagat
     1201 gtccgctttc tggtatgccg tgcgtactgc ggtgatcaac gccgccagcg gtcgtcagac
     1261 tgtcgatgaa gccctgaaag acgcgcagac taattcgagc tcgaacaaca acaacaataa
     1321 caataacaac aacctcggga tcgacgaaaa cctgtacctc acaagtttgt acaaaaaagc
     1381 tgaacgagaa acgtaaaatg atataaatat caatatatta aattagattt tgcataaaaa
     1441 acagactaca taatactgta aaacacaaca tatccagtca ctatggcggc cgcattaggc
     1501 accccaggct ttacacttta tgctttcggc tcgtataatg tgtggatttt gagttaggat
     1561 ccgtcgagat tttcaggagc taaggaagct aaaatggaga aaaaaatcac tggatatacc
     1621 accgttgata tatcccaatg gcatcgtaaa gaacattttg aggcatttca gtcagttgct
     1681 caatgtacct ataaccagac cgttcagctg gatattacgg cctttttaaa gaccgtaaag
     1741 aaaaataagc acaagtttta tccggccttt attcacattc ttgcccgcct gatgaatgct
     1801 catccggaat tccgtatggc aatgaaagac ggtgagctgg tgatatggga tagtgttcac
     1861 ccttgttaca ccgttttcca tgagcaaact gaaacgtttt catcgctctg gagtgaatac
     1921 cacgacgatt tccggcagtt tctacacata tattcgcaag atgtggcgtg ttacggtgaa
     1981 aacctggcct atttccctaa agggtttatt gagaatatgt ttttcgtctc agccaatccc
     2041 tgggtgagtt tcaccagttt tgatttaaac gtggccaata tggacaactt cttcgccccc
     2101 gttttcacca tgggcaaata ttatacgcaa ggcgacaagg tgctgatgcc gctggcgatt
     2161 caggttcatc atgccgtttg tgatggcttc catgtcggca gaatgcttaa tgaattacaa
     2221 cagtactgcg atgagtggca gggcggggcg taaagatctg gatccggctt actaaaagcc
     2281 agataacagt atgcgtattt gcgcgctgat ttttgcggta taagaatata tactgatatg
     2341 tatacccgaa gtatgtcaaa aagaggtatg ctatgaagca gcgtattaca gtgacagttg
     2401 acagcgacag ctatcagttg ctcaaggcat atatgatgtc aatatctccg gtctggtaag
     2461 cacaaccatg cagaatgaag cccgtcgtct gcgtgccgaa cgctggaaag cggaaaatca
     2521 ggaagggatg gctgaggtcg cccggtttat tgaaatgaac ggctcttttg ctgacgagaa
     2581 caggggctgg tgaaatgcag tttaaggttt acacctataa aagagagagc cgttatcgtc
     2641 tgtttgtgga tgtacagagt gatattattg acacgcccgg gcgacggatg gtgatccccc
     2701 tggccagtgc acgtctgctg tcagataaag tctcccgtga actttacccg gtggtgcata
     2761 tcggggatga aagctggcgc atgatgacca ccgatatggc cagtgtgccg gtctccgtta
     2821 tcggggaaga agtggctgat ctcagccacc gcgaaaatga catcaaaaac gccattaacc
     2881 tgatgttctg gggaatataa atgtcaggct cccttataca cagccagtct gcaggtcgac
     2941 catagtgact ggatatgttg tgttttacag tattatgtag tctgtttttt atgcaaaatc
     3001 taatttaata tattgatatt tatatcattt tacgtttctc gttcagcttt cttgtacaaa
     3061 gtggtgatgg cgcgcctgta ggacgtcgac ggtaccatcg atacgcgttc gaagcttaat
     3121 tagctgagct tggactcctg ttgatagatc cagtaatgac ctcagaactc catctggatt
     3181 tgttcagaac gctcggttgc cgccgggcgt tttttattgg tgagaatcca agctagcttg
     3241 gcgagatttt caggagctaa ggaagctaaa atggagaaaa aaatcactgg atataccacc
     3301 gttgatatat cccaatggca tcgtaaagaa cattttgagg catttcagtc agttgctcaa
     3361 tgtacctata accagaccgt tcagctggat attacggcct ttttaaagac cgtaaagaaa
     3421 aataagcaca agttttatcc ggcctttatt cacattcttg cccgcctgat gaatgctcat
     3481 ccggaatttc gtatggcaat gaaagacggt gagctggtga tatgggatag tgttcaccct
     3541 tgttacaccg ttttccatga gcaaactgaa acgttttcat cgctctggag tgaataccac
     3601 gacgatttcc ggcagtttct acacatatat tcgcaagatg tggcgtgtta cggtgaaaac
     3661 ctggcctatt tccctaaagg gtttattgag aatatgtttt tcgtctcagc caatccctgg
     3721 gtgagtttca ccagttttga tttaaacgtg gccaatatgg acaacttctt cgcccccgtt
     3781 ttcaccatgg gcaaatatta tacgcaaggc gacaaggtgc tgatgccgct ggcgattcag
     3841 gttcatcatg ccgtttgtga tggcttccat gtcggcagaa tgcttaatga attacaacag
     3901 tactgcgatg agtggcaggg cggggcgtaa tttttttaag gcagttattg gtgcccttaa
     3961 acgcctgggg taatgactct ctagcttgag gcatcaaata aaacgaaagg ctcagtcgaa
     4021 agactgggcc tttcgtttta tctgttgttt gtcggtgaac gctctcctga gtaggacaaa
     4081 tccgccctct agattacgtg cagtcgatga taagctgtca aacatgagaa ttgtgcctaa
     4141 tgagtgagct aacttacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     4201 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     4261 gggcgccagg gtggtttttc ttttcaccag tgagacgggc aacagctgat tgcccttcac
     4321 cgcctggccc tgagagagtt gcagcaagcg gtccacgctg gtttgcccca gcaggcgaaa
     4381 atcctgtttg atggtggtta acggcgggat ataacatgag ctgtcttcgg tatcgtcgta
     4441 tcccactacc gagatatccg caccaacgcg cagcccggac tcggtaatgg cgcgcattgc
     4501 gcccagcgcc atctgatcgt tggcaaccag catcgcagtg ggaacgatgc cctcattcag
     4561 catttgcatg gtttgttgaa aaccggacat ggcactccag tcgccttccc gttccgctat
     4621 cggctgaatt tgattgcgag tgagatattt atgccagcca gccagacgca gacgcgccga
     4681 gacagaactt aatgggcccg ctaacagcgc gatttgctgg tgacccaatg cgaccagatg
     4741 ctccacgccc agtcgcgtac cgtcttcatg ggagaaaata atactgttga tgggtgtctg
     4801 gtcagagaca tcaagaaata acgccggaac attagtgcag gcagcttcca cagcaatggc
     4861 atcctggtca tccagcggat agttaatgat cagcccactg acgcgttgcg cgagaagatt
     4921 gtgcaccgcc gctttacagg cttcgacgcc gcttcgttct accatcgaca ccaccacgct
     4981 ggcacccagt tgatcggcgc gagatttaat cgccgcgaca atttgcgacg gcgcgtgcag
     5041 ggccagactg gaggtggcaa cgccaatcag caacgactgt ttgcccgcca gttgttgtgc
     5101 cacgcggttg ggaatgtaat tcagctccgc catcgccgct tccacttttt cccgcgtttt
     5161 cgcagaaacg tggctggcct ggttcaccac gcgggaaacg gtctgataag agacaccggc
     5221 atactctgcg acatcgtata acgttactgg tttcacattc accaccctga attgactctc
     5281 ttccgggcgc tatcatgcca taccgcgaaa ggttttgcac cattcgatgg tgtcggaatt
     5341 tcgggcagcg ttgggtcctg gccacgggtg cgcatgatct agagctgcct cgcgcgtttc
     5401 ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg
     5461 taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt
     5521 cggggcgcag ccatgaccca gtcacgtagc gatagcggag tgtatactgg cttaactatg
     5581 cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat
     5641 gcgtaaggag aaaataccgc atcaggcgct cttccgcttc ctcgctcact gactcgctgc
     5701 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     5761 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     5821 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     5881 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     5941 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     6001 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     6061 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     6121 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     6181 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     6241 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat
     6301 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     6361 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     6421 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     6481 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     6541 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     6601 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     6661 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     6721 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     6781 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     6841 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     6901 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     6961 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     7021 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     7081 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     7141 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     7201 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     7261 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     7321 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     7381 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     7441 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     7501 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     7561 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta
     7621 ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt cttca
//
