LOCUS       pET104-DEST.        7618 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Gateway(R) destination vector for high-level inducible expression of
            biotinylated proteins in bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    pET104-DEST
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7618)
  AUTHORS   Invitrogen (Life Technologies)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 7618)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7618
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        209..227
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    228..252
                     /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             267..289
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             297..299
                     /codon_start=1
                     /product="start codon"
                     /label=start codon
                     /note="ATG"
                     /translation="M"
     CDS             300..515
                     /label=BioEase(TM) tag
                     /note="biotinylated sequence derived from the C-terminus of
                     the alpha-subunit of Klebsiella pneumoniae oxaloacetate 
                     decarboxylase"
     CDS             528..551
                     /label=Xpress(TM) tag
                     /note="Xpress(TM) epitope tag, including an enterokinase 
                     recognition and cleavage site"
     protein_bind    561..685
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        710..740
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             794..1450
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     CDS             1774..2076
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     protein_bind    complement(2120..2244)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     terminator      2330..2377
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        2701..2805
                     /label=AmpR promoter
     CDS             2806..3663
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      3837..4425
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(4611..4753)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(4858..5046)
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     protein_bind    complement(6324..6345)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(6361..7440)
                     /label=lacI
                     /note="lac repressor"
     promoter        complement(7441..7518)
                     /label=lacI promoter
ORIGIN
        1 caaggagatg gcgcccaaca gtcccccggc cacggggcct gccaccatac ccacgccgaa
       61 acaagcgctc atgagcccga agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat
      121 ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta
      181 gaggatcgag atctcgatcc cgcgaaatta atacgactca ctatagggga attgtgagcg
      241 gataacaatt cccctctaga aataattttg tttaacttta agaaggagat atacatatgg
      301 gcgccggcac cccggtgacc gccccgctgg cgggcactat ctggaaggtg ctggccagcg
      361 aaggccagac ggtggccgca ggcgaggtgc tgctgattct ggaagccatg aagatggaaa
      421 ccgaaatccg cgccgcgcag gccgggaccg tgcgcggtat cgcggtgaaa gccggcgacg
      481 cggtggcggt cggcgacacc ctgatgaccc tggcgggctc tggatccgat ctgtacgacg
      541 atgacgataa gggaattatc acaagtttgt acaaaaaagc tgaacgagaa acgtaaaatg
      601 atataaatat caatatatta aattagattt tgcataaaaa acagactaca taatactgta
      661 aaacacaaca tatccagtca ctatggcggc cgcattaggc accccaggct ttacacttta
      721 tgcttccggc tcgtataatg tgtggatttt gagttaggat ccggcgagat tttcaggagc
      781 taaggaagct aaaatggaga aaaaaatcac tggatatacc accgttgata tatcccaatg
      841 gcatcgtaaa gaacattttg aggcatttca gtcagttgct caatgtacct ataaccagac
      901 cgttcagctg gatattacgg cctttttaaa gaccgtaaag aaaaataagc acaagtttta
      961 tccggccttt attcacattc ttgcccgcct gatgaatgct catccggaat tccgtatggc
     1021 aatgaaagac ggtgagctgg tgatatggga tagtgttcac ccttgttaca ccgttttcca
     1081 tgagcaaact gaaacgtttt catcgctctg gagtgaatac cacgacgatt tccggcagtt
     1141 tctacacata tattcgcaag atgtggcgtg ttacggtgaa aacctggcct atttccctaa
     1201 agggtttatt gagaatatgt ttttcgtctc agccaatccc tgggtgagtt tcaccagttt
     1261 tgatttaaac gtggccaata tggacaactt cttcgccccc gttttcacca tgggcaaata
     1321 ttatacgcaa ggcgacaagg tgctgatgcc gctggcgatt caggttcatc atgccgtctg
     1381 tgatggcttc catgtcggca gaatgcttaa tgaattacaa cagtactgcg atgagtggca
     1441 gggcggggcg taaacgcgtg gatccggctt actaaaagcc agataacagt atgcgtattt
     1501 gcgcgcaccg gtgctagcgt atacccgaag tatgtcaaaa agaggtgtgc tatgaagcag
     1561 cgtattacag tgacagttga cagcgacagc tatcagttgc tcaaggcata tatgatgtca
     1621 atatctccgg tctggtaagc acaaccatgc agaatgaagc ccgtcgtctg cgtgccgaac
     1681 gctggaaagc ggaaaatcag gaagggatgg ctgaggtcgc ccggtttatt gaaatgaacg
     1741 gctcttttgc tgacgagaac agggactggt gaaatgcagt ttaaggttta cacctataaa
     1801 agagagagcc gttatcgtct gtttgtggat gtacagagtg atattattga cacgcccggg
     1861 cgacggatgg tgatccccct ggccagtgca cgtctgctgt cagataaagt ctcccgtgaa
     1921 ctttacccgg tggtgcatat cggggatgaa agctggcgca tgatgaccac cgatatggcc
     1981 agtgtgccgg tctccgttat cggggaagaa gtggctgatc tcagccaccg cgaaaatgac
     2041 atcaaaaacg ccattaacct gatgttctgg ggaatataaa tgtcaggctc cgttatacac
     2101 agccagtctg caggtcgacc atagtgactg gatatgttgt gttttacagt attatgtagt
     2161 ctgtttttta tgcaaaatct aatttaatat attgatattt atatcatttt acgtttctcg
     2221 ttcagctttc ttgtacaaag tggtgataat taattaagat agctcagatc cggctgctaa
     2281 caaagcccga aaggaagctg agttggctgc tgccaccgct gagcaataac tagcataacc
     2341 ccttggggcc tctaaacggg tcttgagggg ttttttgctg aaaggaggaa ctatatccgg
     2401 atatcccgca agaggcccgg cagtaccggc ataaccaagc ctatgcctac agcatccagg
     2461 gtgacggtgc cgaggatgac gatgagcgca ttgttagatt tcatacacgg tgcctgactg
     2521 cgttagcaat ttaactgtga taaactaccg cattaaagct agcttatcga tgataagctg
     2581 tcaaacatga gaattaattc ttgaagacga aagggcctcg tgatacgcct atttttatag
     2641 gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg
     2701 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     2761 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     2821 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     2881 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     2941 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     3001 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg
     3061 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     3121 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     3181 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     3241 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     3301 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgc agcaatggca
     3361 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     3421 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     3481 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     3541 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     3601 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     3661 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     3721 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     3781 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     3841 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     3901 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     3961 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
     4021 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     4081 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     4141 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     4201 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     4261 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     4321 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     4381 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     4441 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     4501 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     4561 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg
     4621 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatatggtgc actctcagta
     4681 caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg
     4741 ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     4801 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     4861 gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc
     4921 gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag
     4981 aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt
     5041 ggtcactgat gcctccgtgt aagggggatt tctgttcatg ggggtaatga taccgatgaa
     5101 acgagagagg atgctcacga tacgggttac tgatgatgaa catgcccggt tactggaacg
     5161 ttgtgagggt aaacaactgg cggtatggat gcggcgggac cagagaaaaa tcactcaggg
     5221 tcaatgccag cgcttcgtta atacagatgt aggtgttcca cagggtagcc agcagcatcc
     5281 tgcgatgcag atccggaaca taatggtgca gggcgctgac ttccgcgttt ccagacttta
     5341 cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg ttttgcagca
     5401 gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc tgctaaccag taaggcaacc
     5461 ccgccagcct agccgggtcc tcaacgacag gagcacgatc atgcgcaccc gtggccagga
     5521 cccaacgctg cccgagatgc gccgcgtgcg gctgctggag atggcggacg cgatggatat
     5581 gttctgccaa gggttggttt gcgcattcac agttctccgc aagaattgat tggctccaat
     5641 tcttggagtg gtgaatccgt tagcgaggtg ccgccggctt ccattcaggt cgaggtggcc
     5701 cggctccatg caccgcgacg caacgcgggg aggcagacaa ggtatagggc ggcgcctaca
     5761 atccatgcca acccgttcca tgtgctcgcc gaggcggcat aaatcgccgt gacgatcagc
     5821 ggtccagtga tcgaagttag gctggtaaga gccgcgagcg atccttgaag ctgtccctga
     5881 tggtcgtcat ctacctgcct ggacagcatg gcctgcaacg cgggcatccc gatgccgccg
     5941 gaagcgagaa gaatcataat ggggaaggcc atccagcctc gcgtcgcgaa cgccagcaag
     6001 acgtagccca gcgcgtcggc cgccatgccg gcgataatgg cctgcttctc gccgaaacgt
     6061 ttggtggcgg gaccagtgac gaaggcttga gcgagggcgt gcaagattcc gaataccgca
     6121 agcgacaggc cgatcatcgt cgcgctccag cgaaagcggt cctcgccgaa aatgacccag
     6181 agcgctgccg gcacctgtcc tacgagttgc atgataaaga agacagtcat aagtgcggcg
     6241 acgatagtca tgccccgcgc ccaccggaag gagctgactg ggttgaaggc tctcaagggc
     6301 atcggtcgag atcccggtgc ctaatgagtg agctaactta cattaattgc gttgcgctca
     6361 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     6421 gcggggagag gcggtttgcg tattgggcgc cagggtggtt tttcttttca ccagtgagac
     6481 gggcaacagc tgattgccct tcaccgcctg gccctgagag agttgcagca agcggtccac
     6541 gctggtttgc cccagcaggc gaaaatcctg tttgatggtg gttaacggcg ggatataaca
     6601 tgagctgtct tcggtatcgt cgtatcccac taccgagata tccgcaccaa cgcgcagccc
     6661 ggactcggta atggcgcgca ttgcgcccag cgccatctga tcgttggcaa ccagcatcgc
     6721 agtgggaacg atgccctcat tcagcatttg catggtttgt tgaaaaccgg acatggcact
     6781 ccagtcgcct tcccgttccg ctatcggctg aatttgattg cgagtgagat atttatgcca
     6841 gccagccaga cgcagacgcg ccgagacaga acttaatggg cccgctaaca gcgcgatttg
     6901 ctggtgaccc aatgcgacca gatgctccac gcccagtcgc gtaccgtctt catgggagaa
     6961 aataatactg ttgatgggtg tctggtcaga gacatcaaga aataacgccg gaacattagt
     7021 gcaggcagct tccacagcaa tggcatcctg gtcatccagc ggatagttaa tgatcagccc
     7081 actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta caggcttcga cgccgcttcg
     7141 ttctaccatc gacaccacca cgctggcacc cagttgatcg gcgcgagatt taatcgccgc
     7201 gacaatttgc gacggcgcgt gcagggccag actggaggtg gcaacgccaa tcagcaacga
     7261 ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg taattcagct ccgccatcgc
     7321 cgcttccact ttttcccgcg ttttcgcaga aacgtggctg gcctggttca ccacgcggga
     7381 aacggtctga taagagacac cggcatactc tgcgacatcg tataacgtta ctggtttcac
     7441 attcaccacc ctgaattgac tctcttccgg gcgctatcat gccataccgc gaaaggtttt
     7501 gcgccattcg atggtgtccg ggatctcgac gctctccctt atgcgactcc tgcattagga
     7561 agcagcccag tagtaggttg aggccgttga gcaccgccgc cgcaaggaat ggtgcatg
//
