LOCUS       pCOLA-2-DEST.        5131 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Gateway(R) Nova bacterial destination vector with a ColA origin for 
            expressing proteins tagged at the C-terminus with Strep-Tag II.
ACCESSION   .
VERSION     .
KEYWORDS    pCOLA-2-DEST
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5131)
  AUTHORS   Novagen (EMD Millipore)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 5131)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5131
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(227..1039)
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     promoter        complement(1040..1131)
                     /label=AmpR promoter
     rep_origin      complement(1149..1784)
                     /direction=LEFT
                     /label=ColA ori
                     /note="Plasmids containing the ColA origin of replication
                     can be propagated in E. coli cells that contain additional 
                     plasmids with compatible origins."
     protein_bind    complement(1948..1969)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(1985..3064)
                     /label=lacI
                     /note="lac repressor"
     promoter        complement(3065..3142)
                     /label=lacI promoter
     promoter        3188..3206
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3207..3231
                     /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             3246..3268
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3275..3277
                     /codon_start=1
                     /product="start codon"
                     /label=start codon
                     /note="ATG"
                     /translation="M"
     CDS             3281..3298
                     /label=6xHis
                     /note="6xHis affinity tag"
     protein_bind    3302..3426
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        3451..3481
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             3535..4191
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     CDS             4536..4838
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     protein_bind    complement(4882..5006)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             5014..5037
                     /label=Strep-Tag II
                     /note="peptide that binds Strep-Tactin(R), an engineered 
                     form
                     of streptavidin"
     terminator      5078..5125
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 cctcaggcat ttgagaagca cacggtcaca ctgcttccgg tagtcaataa accggtaaac
       61 cagcaataga cataagcggc tatttaacga ccctgccctg aaccgacgac aagctgacga
      121 ccgggtctcc gcaagtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
      181 tttctaaata cattcaaata tgtatccgct catgaattaa ttcttagaaa aactcatcga
      241 gcatcaaatg aaactgcaat ttattcatat caggattatc aataccatat ttttgaaaaa
      301 gccgtttctg taatgaagga gaaaactcac cgaggcagtt ccataggatg gcaagatcct
      361 ggtatcggtc tgcgattccg actcgtccaa catcaataca acctattaat ttcccctcgt
      421 caaaaataag gttatcaagt gagaaatcac catgagtgac gactgaatcc ggtgagaatg
      481 gcaaaagttt atgcatttct ttccagactt gttcaacagg ccagccatta cgctcgtcat
      541 caaaatcact cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga gcgagacgaa
      601 atacgcggtc gctgttaaaa ggacaattac aaacaggaat cgaatgcaac cggcgcagga
      661 acactgccag cgcatcaaca atattttcac ctgaatcagg atattcttct aatacctgga
      721 atgctgtttt cccggggatc gcagtggtga gtaaccatgc atcatcagga gtacggataa
      781 aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg accatctcat
      841 ctgtaacatc attggcaacg ctacctttgc catgtttcag aaacaactct ggcgcatcgg
      901 gcttcccata caatcgatag attgtcgcac ctgattgccc gacattatcg cgagcccatt
      961 tatacccata taaatcagca tccatgttgg aatttaatcg cggcctagag caagacgttt
     1021 cccgttgaat atggctcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     1081 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggcatgc
     1141 tagcgcagaa acgtcctaga agatgccagg aggatactta gcagagagac aataaggccg
     1201 gagcgaagcc gtttttccat aggctccgcc cccctgacga acatcacgaa atctgacgct
     1261 caaatcagtg gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctgatg
     1321 gctccctctt gcgctctcct gttcccgtcc tgcggcgtcc gtgttgtggt ggaggcttta
     1381 cccaaatcac cacgtcccgt tccgtgtaga cagttcgctc caagctgggc tgtgtgcaag
     1441 aaccccccgt tcagcccgac tgctgcgcct tatccggtaa ctatcatctt gagtccaacc
     1501 cggaaagaca cgacaaaacg ccactggcag cagccattgg taactgagaa ttagtggatt
     1561 tagatatcga gagtcttgaa gtggtggcct aacagaggct acactgaaag gacagtattt
     1621 ggtatctgcg ctccactaaa gccagttacc aggttaagca gttccccaac tgacttaacc
     1681 ttcgatcaaa ccgcctcccc aggcggtttt ttcgtttaca gagcaggaga ttacgacgat
     1741 cgtaaaagga tctcaagaag atcctttacg gattcccgac accatcactc tagatttcag
     1801 tgcaatttat ctcttcaaat gtagcacctg aagtcagccc catacgatat aagttgtaat
     1861 tctcatgtta gtcatgcccc gcgcccaccg gaaggagctg actgggttga aggctctcaa
     1921 gggcatcggt cgagatcccg gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg
     1981 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     2041 acgcgcgggg agaggcggtt tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg
     2101 agacgggcaa cagctgattg cccttcaccg cctggccctg agagagttgc agcaagcggt
     2161 ccacgctggt ttgccccagc aggcgaaaat cctgtttgat ggtggttaac ggcgggatat
     2221 aacatgagct gtcttcggta tcgtcgtatc ccactaccga gatgtccgca ccaacgcgca
     2281 gcccggactc ggtaatggcg cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca
     2341 tcgcagtggg aacgatgccc tcattcagca tttgcatggt ttgttgaaaa ccggacatgg
     2401 cactccagtc gccttcccgt tccgctatcg gctgaatttg attgcgagtg agatatttat
     2461 gccagccagc cagacgcaga cgcgccgaga cagaacttaa tgggcccgct aacagcgcga
     2521 tttgctggtg acccaatgcg accagatgct ccacgcccag tcgcgtaccg tcttcatggg
     2581 agaaaataat actgttgatg ggtgtctggt cagagacatc aagaaataac gccggaacat
     2641 tagtgcaggc agcttccaca gcaatggcat cctggtcatc cagcggatag ttaatgatca
     2701 gcccactgac gcgttgcgcg agaagattgt gcaccgccgc tttacaggct tcgacgccgc
     2761 ttcgttctac catcgacacc accacgctgg cacccagttg atcggcgcga gatttaatcg
     2821 ccgcgacaat ttgcgacggc gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca
     2881 acgactgttt gcccgccagt tgttgtgcca cgcggttggg aatgtaattc agctccgcca
     2941 tcgccgcttc cactttttcc cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc
     3001 gggaaacggt ctgataagag acaccggcat actctgcgac atcgtataac gttactggtt
     3061 tcacattcac caccctgaat tgactctctt ccgggcgcta tcatgccata ccgcgaaagg
     3121 ttttgcgcca ttcgatggtg tccgggatct cgacgctctc ccttatgcga ctcctgcatt
     3181 aggaaattaa tacgactcac tataggggaa ttgtgagcgg ataacaattc ccctgtagaa
     3241 ataattttgt ttaactttaa taaggagata taccatggca catcaccacc accatcacgt
     3301 cacaagtttg tacaaaaaag ctgaacgaga aacgtaaaat gatataaata tcaatatatt
     3361 aaattagatt ttgcataaaa aacagactac ataatactgt aaaacacaac atatccagtc
     3421 actatggcgg ccgcattagg caccccaggc tttacacttt atgcttccgg ctcgtataat
     3481 gtgtggattt tgagttagga tccgtcgaga ttttcaggag ctaaggaagc taaaatggag
     3541 aaaaaaatca ctggatatac caccgttgat atatcccaat ggcatcgtaa agaacatttt
     3601 gaggcatttc agtcagttgc tcaatgtacc tataaccaga ccgttcagct ggatattacg
     3661 gcctttttaa agaccgtaaa gaaaaataag cacaagtttt atccggcctt tattcacatt
     3721 cttgcccgcc tgatgaatgc tcatccggaa ttccgtatgg caatgaaaga cggtgagctg
     3781 gtgatatggg atagtgttca cccttgttac accgttttcc atgagcaaac tgaaacgttt
     3841 tcatcgctct ggagtgaata ccacgacgat ttccggcagt ttctacacat atattcgcaa
     3901 gatgtggcgt gttacggtga aaacctggcc tatttcccta aagggtttat tgagaatatg
     3961 tttttcgtct cagccaatcc ctgggtgagt ttcaccagtt ttgatttaaa cgtggccaat
     4021 atggacaact tcttcgcccc cgttttcact atgggcaaat attatacgca aggcgacaag
     4081 gtgctgatgc cgctggcgat tcaggttcat catgccgttt gtgatggctt ccatgtcggc
     4141 agaatgctta atgaattaca acagtactgc gatgagtggc agggcggggc gtaaacgcgt
     4201 ggatccggct tactaaaagc cagataacag tatgcgtatt tgcgcgctga tttttgcggt
     4261 ataagaatat atactgatat gtatacccga agtatgtcaa aaagaggtat gctatgaagc
     4321 agcgtattac agtgacagtt gacagcgaca gctatcagtt gctcaaggca tatatgatgt
     4381 caatatctcc ggtctggtaa gcacaaccat gcagaatgaa gcccgtcgtc tgcgtgccga
     4441 acgctggaaa gcggaaaatc aggaagggat ggctgaggtc gcccggttta ttgaaatgaa
     4501 cggctctttt gctgacgaga acaggggctg gtgaaatgca gtttaaggtt tacacctata
     4561 aaagagagag ccgttatcgt ctgtttgtgg atgtacagag tgatattatt gacacgcccg
     4621 ggcgacggat ggtgatcccc ctggccagtg cacgtctgct gtcagataaa gtctcccgtg
     4681 aactttaccc ggtggtgcat atcggggatg aaagctggcg catgatgacc accgatatgg
     4741 ccagtgtgcc ggtttccgtt atcggggaag aagtggctga tctcagccac cgcgaaaatg
     4801 acatcaaaaa cgccattaac ctgatgttct ggggaatata aatgtcaggc tcccttatac
     4861 acagccagtc tgcaggtcga ccatagtgac tggatatgtt gtgttttaca gtattatgta
     4921 gtctgttttt tatgcaaaat ctaatttaat atattgatat ttatatcatt ttacgtttct
     4981 cgttcagctt tcttgtacaa agtggtttcc gcttggagcc acccgcagtt cgaaaagtaa
     5041 gtgattaacc taggctgctg ccaccgctga gcaataacta gcataacccc ttggggcctc
     5101 taaacgggtc ttgaggggtt ttttgctgaa a
//
