LOCUS       pET160-DEST.        7437 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Gateway(R) destination vector for high-level bacterial expression of
            proteins with an N-terminal tetracysteine (Lumio(TM)) tag.
ACCESSION   .
VERSION     .
KEYWORDS    pET160-DEST
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7437)
  AUTHORS   Invitrogen (Life Technologies)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 7437)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7437
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        21..39
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    40..64
                     /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             79..101
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             109..111
                     /codon_start=1
                     /product="start codon"
                     /label=start codon
                     /note="ATG"
                     /translation="M"
     CDS             112..129
                     /label=6xHis
                     /note="6xHis affinity tag"
     CDS             142..159
                     /label=tetracysteine tag
                     /note="tetracysteine peptide that binds biarsenical
                     labeling reagents"
     CDS             169..189
                     /label=TEV site
                     /note="tobacco etch virus (TEV) protease recognition and 
                     cleavage site"
     protein_bind    196..320
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        345..375
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             429..1085
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     CDS             1409..1711
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     protein_bind    complement(1755..1879)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     terminator      1961..2008
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        2332..2436
                     /label=AmpR promoter
     CDS             2437..3294
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      3468..4056
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(4242..4384)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(4489..4677)
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     protein_bind    complement(5955..5976)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(5992..7071)
                     /label=lacI
                     /note="lac repressor"
     promoter        complement(7072..7149)
                     /label=lacI promoter
ORIGIN
        1 agatctcgat cccgcgaaat taatacgact cactataggg gaattgtgag cggataacaa
       61 ttcccctcta gaaataattt tgtttaactt taagaaggag atatacatat gcatcatcac
      121 catcaccatg gtgctggtgg ctgttgtcct ggctgttgcg gtggcggcga aaacctgtat
      181 tttcagggaa ttatcacaag tttgtacaaa aaagctgaac gagaaacgta aaatgatata
      241 aatatcaata tattaaatta gattttgcat aaaaaacaga ctacataata ctgtaaaaca
      301 caacatatcc agtcactatg gcggccgcat taggcacccc aggctttaca ctttatgctt
      361 ccggctcgta taatgtgtgg attttgagtt aggatccggc gagattttca ggagctaagg
      421 aagctaaaat ggagaaaaaa atcactggat ataccaccgt tgatatatcc caatggcatc
      481 gtaaagaaca ttttgaggca tttcagtcag ttgctcaatg tacctataac cagaccgttc
      541 agctggatat tacggccttt ttaaagaccg taaagaaaaa taagcacaag ttttatccgg
      601 cctttattca cattcttgcc cgcctgatga atgctcatcc ggaattccgt atggcaatga
      661 aagacggtga gctggtgata tgggatagtg ttcacccttg ttacaccgtt ttccatgagc
      721 aaactgaaac gttttcatcg ctctggagtg aataccacga cgatttccgg cagtttctac
      781 acatatattc gcaagatgtg gcgtgttacg gtgaaaacct ggcctatttc cctaaagggt
      841 ttattgagaa tatgtttttc gtctcagcca atccctgggt gagtttcacc agttttgatt
      901 taaacgtggc caatatggac aacttcttcg cccccgtttt caccatgggc aaatattata
      961 cgcaaggcga caaggtgctg atgccgctgg cgattcaggt tcatcatgcc gtctgtgatg
     1021 gcttccatgt cggcagaatg cttaatgaat tacaacagta ctgcgatgag tggcagggcg
     1081 gggcgtaaac gcgtggatcc ggcttactaa aagccagata acagtatgcg tatttgcgcg
     1141 caccggtgct agcgtatacc cgaagtatgt caaaaagagg tgtgctatga agcagcgtat
     1201 tacagtgaca gttgacagcg acagctatca gttgctcaag gcatatatga tgtcaatatc
     1261 tccggtctgg taagcacaac catgcagaat gaagcccgtc gtctgcgtgc cgaacgctgg
     1321 aaagcggaaa atcaggaagg gatggctgag gtcgcccggt ttattgaaat gaacggctct
     1381 tttgctgacg agaacaggga ctggtgaaat gcagtttaag gtttacacct ataaaagaga
     1441 gagccgttat cgtctgtttg tggatgtaca gagtgatatt attgacacgc ccgggcgacg
     1501 gatggtgatc cccctggcca gtgcacgtct gctgtcagat aaagtctccc gtgaacttta
     1561 cccggtggtg catatcgggg atgaaagctg gcgcatgatg accaccgata tggccagtgt
     1621 gccggtctcc gttatcgggg aagaagtggc tgatctcagc cgccgcgaaa atgacatcaa
     1681 aaacgccatt aacctgatgt tctggggaat ataaatgtca ggctccctta tacacagcca
     1741 gtctgcaggt cgaccatagt gactggatat gttgtgtttt acagtattat gtagtctgtt
     1801 ttttatgcaa aatctaattt aatatattga tatttatatc attttacgtt tctcgttcag
     1861 ctttcttgta caaagtggtg ataattaatt aagatcagat ccggctgcta acaaagcccg
     1921 aaaggaagct gagttggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc
     1981 ctctaaacgg gtcttgaggg gttttttgct gaaaggagga actatatccg gatatcccgc
     2041 aagaggcccg gcagtaccgg cataaccaag cctatgccta cagcatccag ggtgacggtg
     2101 ccgaggatga cgatgagcgc attgttagat ttcatacacg gtgcctgact gcgttagcaa
     2161 tttaactgtg ataaactacc gcattaaagc tagcttatcg atgataagct gtcaaacatg
     2221 agaattaatt cttgaagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
     2281 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     2341 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     2401 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     2461 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     2521 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     2581 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     2641 acttttaaag ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa
     2701 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     2761 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     2821 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     2881 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     2941 gaagccatac caaacgacga gcgtgacacc acgatgcctg cagcaatggc aacaacgttg
     3001 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     3061 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     3121 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     3181 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     3241 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     3301 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     3361 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     3421 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     3481 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     3541 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     3601 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     3661 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     3721 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     3781 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     3841 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     3901 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     3961 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     4021 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     4081 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     4141 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     4201 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc
     4261 cttacgcatc tgtgcggtat ttcacaccgc atatatggtg cactctcagt acaatctgct
     4321 ctgatgccgc atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc
     4381 tgcgccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     4441 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     4501 gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga
     4561 ttcacagatg tctgcctgtt catccgcgtc cagctcgttg agtttctcca gaagcgttaa
     4621 tgtctggctt ctgataaagc gggccatgtt aagggcggtt ttttcctgtt tggtcactga
     4681 tgcctccgtg taagggggat ttctgttcat gggggtaatg ataccgatga aacgagagag
     4741 gatgctcacg atacgggtta ctgatgatga acatgcccgg ttactggaac gttgtgaggg
     4801 taaacaactg gcggtatgga tgcggcggga ccagagaaaa atcactcagg gtcaatgcca
     4861 gcgcttcgtt aatacagatg taggtgttcc acagggtagc cagcagcatc ctgcgatgca
     4921 gatccggaac ataatggtgc agggcgctga cttccgcgtt tccagacttt acgaaacacg
     4981 gaaaccgaag accattcatg ttgttgctca ggtcgcagac gttttgcagc agcagtcgct
     5041 tcacgttcgc tcgcgtatcg gtgattcatt ctgctaacca gtaaggcaac cccgccagcc
     5101 tagccgggtc ctcaacgaca ggagcacgat catgcgcacc cgtggccagg acccaacgct
     5161 gcccgagatg cgccgcgtgc ggctgctgga gatggcggac gcgatggata tgttctgcca
     5221 agggttggtt tgcgcattca cagttctccg caagaattga ttggctccaa ttcttggagt
     5281 ggtgaatccg ttagcgaggt gccgccggct tccattcagg tcgaggtggc ccggctccat
     5341 gcaccgcgac gcaacgcggg gaggcagaca aggtataggg cggcgcctac aatccatgcc
     5401 aacccgttcc atgtgctcgc cgaggcggca taaatcgccg tgacgatcag cggtccagtg
     5461 atcgaagtta ggctggtaag agccgcgagc gatccttgaa gctgtccctg atggtcgtca
     5521 tctacctgcc tggacagcat ggcctgcaac gcgggcatcc cgatgccgcc ggaagcgaga
     5581 agaatcataa tggggaaggc catccagcct cgcgtcgcga acgccagcaa gacgtagccc
     5641 agcgcgtcgg ccgccatgcc ggcgataatg gcctgcttct cgccgaaacg tttggtggcg
     5701 ggaccagtga cgaaggcttg agcgagggcg tgcaagattc cgaataccgc aagcgacagg
     5761 ccgatcatcg tcgcgctcca gcgaaagcgg tcctcgccga aaatgaccca gagcgctgcc
     5821 ggcacctgtc ctacgagttg catgataaag aagacagtca taagtgcggc gacgatagtc
     5881 atgccccgcg cccaccggaa ggagctgact gggttgaagg ctctcaaggg catcggtcga
     5941 gatcccggtg cctaatgagt gagctaactt acattaattg cgttgcgctc actgcccgct
     6001 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     6061 ggcggtttgc gtattgggcg ccagggtggt ttttcttttc accagtgaga cgggcaacag
     6121 ctgattgccc ttcaccgcct ggccctgaga gagttgcagc aagcggtcca cgctggtttg
     6181 ccccagcagg cgaaaatcct gtttgatggt ggttaacggc gggatataac atgagctgtc
     6241 ttcggtatcg tcgtatccca ctaccgagat atccgcacca acgcgcagcc cggactcggt
     6301 aatggcgcgc attgcgccca gcgccatctg atcgttggca accagcatcg cagtgggaac
     6361 gatgccctca ttcagcattt gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc
     6421 ttcccgttcc gctatcggct gaatttgatt gcgagtgaga tatttatgcc agccagccag
     6481 acgcagacgc gccgagacag aacttaatgg gcccgctaac agcgcgattt gctggtgacc
     6541 caatgcgacc agatgctcca cgcccagtcg cgtaccgtct tcatgggaga aaataatact
     6601 gttgatgggt gtctggtcag agacatcaag aaataacgcc ggaacattag tgcaggcagc
     6661 ttccacagca atggcatcct ggtcatccag cggatagtta atgatcagcc cactgacgcg
     6721 ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg acgccgcttc gttctaccat
     6781 cgacaccacc acgctggcac ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg
     6841 cgacggcgcg tgcagggcca gactggaggt ggcaacgcca atcagcaacg actgtttgcc
     6901 cgccagttgt tgtgccacgc ggttgggaat gtaattcagc tccgccatcg ccgcttccac
     6961 tttttcccgc gttttcgcag aaacgtggct ggcctggttc accacgcggg aaacggtctg
     7021 ataagagaca ccggcatact ctgcgacatc gtataacgtt actggtttca cattcaccac
     7081 cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt tgcgccattc
     7141 gatggtgtcc gggatctcga cgctctccct tatgcgactc ctgcattagg aagcagccca
     7201 gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc aaggagatgg
     7261 cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa caagcgctca
     7321 tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata taggcgccag
     7381 caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag aggatcg
//
