LOCUS       pET-59-DEST.        7198 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Gateway(R) Nova bacterial destination vector for expressing proteins
            tagged at the N-terminus with thioredoxin and at the C-terminus with
            Strep-Tag II.
ACCESSION   .
VERSION     .
KEYWORDS    pET-59-DEST
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7198)
  AUTHORS   Novagen (EMD Millipore)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 7198)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7198
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      12..467
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             complement(585..1442)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(1443..1535)
                     /label=AmpR promoter
     rep_origin      1616..2204
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(2390..2532)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(2637..2825)
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     protein_bind    complement(3363..3384)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(3400..4479)
                     /label=lacI
                     /note="lac repressor"
     promoter        complement(4480..4557)
                     /label=lacI promoter
     promoter        4870..4888
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    4889..4913
                     /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)."
     RBS             4928..4950
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             4957..4959
                     /codon_start=1
                     /product="start codon"
                     /label=start codon
                     /note="ATG"
                     /translation="M"
     CDS             4969..4986
                     /label=6xHis
                     /note="6xHis affinity tag"
     CDS             4999..5322
                     /label=TrxA
                     /note="E. coli thioredoxin"
     CDS             5329..5346
                     /label=thrombin site
                     /note="thrombin recognition and cleavage site"
     protein_bind    5350..5474
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        5499..5529
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             5583..6239
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     CDS             6584..6886
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     protein_bind    complement(6930..7054)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             7062..7085
                     /label=Strep-Tag II
                     /note="peptide that binds Strep-Tactin(R), an engineered 
                     form
                     of streptavidin"
     terminator      7126..7173
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
       61 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
      121 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
      181 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
      241 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
      301 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
      361 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
      421 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaatttctg gcggcacgat
      481 ggcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
      541 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
      601 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
      661 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
      721 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
      781 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
      841 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
      901 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
      961 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     1021 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     1081 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     1141 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     1201 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     1261 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     1321 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     1381 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     1441 atactcttcc tttttcaatc atgattgaag catttatcag ggttattgtc tcatgagcgg
     1501 atacatattt gaatgtattt agaaaaataa acaaataggt catgaccaaa atcccttaac
     1561 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     1621 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     1681 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     1741 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
     1801 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     1861 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     1921 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     1981 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     2041 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     2101 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     2161 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     2221 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     2281 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     2341 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt
     2401 attttctcct tacgcatctg tgcggtattt cacaccgcat atatggtgca ctctcagtac
     2461 aatctgctct gatgccgcat agttaagcca gtatacactc cgctatcgct acgtgactgg
     2521 gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
     2581 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
     2641 ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg
     2701 tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga
     2761 agcgttaatg tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg
     2821 gtcactgatg cctccgtgta agggggattt ctgttcatgg gggtaatgat accgatgaaa
     2881 cgagagagga tgctcacgat acgggttact gatgatgaac atgcccggtt actggaacgt
     2941 tgtgagggta aacaactggc ggtatggatg cggcgggacc agagaaaaat cactcagggt
     3001 caatgccagc gcttcgttaa tacagatgta ggtgttccac agggtagcca gcagcatcct
     3061 gcgatgcaga tccggaacat aatggtgcag ggcgctgact tccgcgtttc cagactttac
     3121 gaaacacgga aaccgaagac cattcatgtt gttgctcagg tcgcagacgt tttgcagcag
     3181 cagtcgcttc acgttcgctc gcgtatcggt gattcattct gctaaccagt aaggcaaccc
     3241 cgccagccta gccgggtcct caacgacagg agcacgatca tgctagtcat gccccgcgcc
     3301 caccggaagg agctgactgg gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc
     3361 taatgagtga gctaacttac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     3421 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     3481 attgggcgcc agggtggttt ttcttttcac cagtgagacg ggcaacagct gattgccctt
     3541 caccgcctgg ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg
     3601 aaaatcctgt ttgatggtgg ttaacggcgg gatataacat gagctgtctt cggtatcgtc
     3661 gtatcccact accgagatgt ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat
     3721 tgcgcccagc gccatctgat cgttggcaac cagcatcgca gtgggaacga tgccctcatt
     3781 cagcatttgc atggtttgtt gaaaaccgga catggcactc cagtcgcctt cccgttccgc
     3841 tatcggctga atttgattgc gagtgagata tttatgccag ccagccagac gcagacgcgc
     3901 cgagacagaa cttaatgggc ccgctaacag cgcgatttgc tggtgaccca atgcgaccag
     3961 atgctccacg cccagtcgcg taccgtcttc atgggagaaa ataatactgt tgatgggtgt
     4021 ctggtcagag acatcaagaa ataacgccgg aacattagtg caggcagctt ccacagcaat
     4081 ggcatcctgg tcatccagcg gatagttaat gatcagccca ctgacgcgtt gcgcgagaag
     4141 attgtgcacc gccgctttac aggcttcgac gccgcttcgt tctaccatcg acaccaccac
     4201 gctggcaccc agttgatcgg cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg
     4261 cagggccaga ctggaggtgg caacgccaat cagcaacgac tgtttgcccg ccagttgttg
     4321 tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc gcttccactt tttcccgcgt
     4381 tttcgcagaa acgtggctgg cctggttcac cacgcgggaa acggtctgat aagagacacc
     4441 ggcatactct gcgacatcgt ataacgttac tggtttcaca ttcaccaccc tgaattgact
     4501 ctcttccggg cgctatcatg ccataccgcg aaaggttttg cgccattcga tggtgtccgg
     4561 gatctcgacg ctctccctta tgcgactcct gcattaggaa gcagcccagt agtaggttga
     4621 ggccgttgag caccgccgcc gcaaggaatg gtgcatgcaa ggagatggcg cccaacagtc
     4681 ccccggccac ggggcctgcc accataccca cgccgaaaca agcgctcatg agcccgaagt
     4741 ggcgagcccg atcttcccca tcggtgatgt cggcgatata ggcgccagca accgcacctg
     4801 tggcgccggt gatgccggcc acgatgcgtc cggcgtagag gatcgagatc gatctcgatc
     4861 ccgcgaaatt aatacgactc actatagggg aattgtgagc ggataacaat tcccctctag
     4921 aaataatttt gtttaacttt aagaaggaga tataccatgg gcagcagcca tcatcatcat
     4981 catcacagcc atatggctag cgataaaatt attcacctga ctgacgacag ttttgacacg
     5041 gatgtactca aagcggacgg ggcgatcctc gtcgatttct gggcagagtg gtgcggtccg
     5101 tgcaaaatga tcgccccgat tctggatgaa atcgctgacg aatatcaggg caaactgacc
     5161 gttgcaaaac tgaacatcga tcaaaaccct ggcactgcgc cgaaatatgg catccgtggt
     5221 atcccgactc tgctgctgtt caaaaacggt gaagtggcgg caaccaaagt gggtgcactg
     5281 tctaaaggtc agttgaaaga gttcctcgac gctaacctgg cccgagctct tgttccacgt
     5341 ggttctgtca caagtttgta caaaaaagct gaacgagaaa cgtaaaatga tataaatatc
     5401 aatatattaa attagatttt gcataaaaaa cagactacat aatactgtaa aacacaacat
     5461 atccagtcac tatggcggcc gcattaggca ccccaggctt tacactttat gcttccggct
     5521 cgtataatgt gtggattttg agttaggatc cgtcgagatt ttcaggagct aaggaagcta
     5581 aaatggagaa aaaaatcact ggatatacca ccgttgatat atcccaatgg catcgtaaag
     5641 aacattttga ggcatttcag tcagttgctc aatgtaccta taaccagacc gttcagctgg
     5701 atattacggc ctttttaaag accgtaaaga aaaataagca caagttttat ccggccttta
     5761 ttcacattct tgcccgcctg atgaatgctc atccggaatt ccgtatggca atgaaagacg
     5821 gtgagctggt gatatgggat agtgttcacc cttgttacac cgttttccat gagcaaactg
     5881 aaacgttttc atcgctctgg agtgaatacc acgacgattt ccggcagttt ctacacatat
     5941 attcgcaaga tgtggcgtgt tacggtgaaa acctggccta tttccctaaa gggtttattg
     6001 agaatatgtt tttcgtctca gccaatccct gggtgagttt caccagtttt gatttaaacg
     6061 tggccaatat ggacaacttc ttcgcccccg ttttcactat gggcaaatat tatacgcaag
     6121 gcgacaaggt gctgatgccg ctggcgattc aggttcatca tgccgtttgt gatggcttcc
     6181 atgtcggcag aatgcttaat gaattacaac agtactgcga tgagtggcag ggcggggcgt
     6241 aaacgcgtgg atccggctta ctaaaagcca gataacagta tgcgtatttg cgcgctgatt
     6301 tttgcggtat aagaatatat actgatatgt atacccgaag tatgtcaaaa agaggtatgc
     6361 tatgaagcag cgtattacag tgacagttga cagcgacagc tatcagttgc tcaaggcata
     6421 tatgatgtca atatctccgg tctggtaagc acaaccatgc agaatgaagc ccgtcgtctg
     6481 cgtgccgaac gctggaaagc ggaaaatcag gaagggatgg ctgaggtcgc ccggtttatt
     6541 gaaatgaacg gctcttttgc tgacgagaac aggggctggt gaaatgcagt ttaaggttta
     6601 cacctataaa agagagagcc gttatcgtct gtttgtggat gtacagagtg atattattga
     6661 cacgcccggg cgacggatgg tgatccccct ggccagtgca cgtctgctgt cagataaagt
     6721 ctcccgtgaa ctttacccgg tggtgcatat cggggatgaa agctggcgca tgatgaccac
     6781 cgatatggcc agtgtgccgg tttccgttat cggggaagaa gtggctgatc tcagccaccg
     6841 cgaaaatgac atcaaaaacg ccattaacct gatgttctgg ggaatataaa tgtcaggctc
     6901 ccttatacac agccagtctg caggtcgacc atagtgactg gatatgttgt gttttacagt
     6961 attatgtagt ctgtttttta tgcaaaatct aatttaatat attgatattt atatcatttt
     7021 acgtttctcg ttcagctttc ttgtacaaag tggtttccgc ttggagccac ccgcagttcg
     7081 aaaagtaagt gattaaccta ggctgctgcc accgctgagc aataactagc ataacccctt
     7141 ggggcctcta aacgggtctt gaggggtttt ttgctgaaag gaggaactat atccggat
//
