LOCUS       Gateway_Nova_pCOLA-3-DEST   5134 bp    DNA   circular            24-AUG-2007
DEFINITION  Gateway Nova pCOLA-3-DEST  Strep-Tag II/attR1-CamccdB-attR2/6xHis-Tag.
SOURCE      
  ORGANISM  
FEATURES             Location/Qualifiers
     CDS             complement(224..1039)
                     /vntifkey="4"
                     /label=Kan
     CDS             complement(1982..3064)
                     /vntifkey="4"
                     /label=lacI
                     /note="Wildtype lacI gene"
     promoter        3188..3204
                     /vntifkey="30"
                     /label=T7\Promoter
     protein_bind    3209..3229
                     /vntifkey="31"
                     /label=lacO
     RBS             3262..3267
                     /vntifkey="32"
                     /label=RBS-1
     primer          complement(5067..5085)
                     /vntifkey="27"
                     /label=T7\Terminator\primer
     terminator      5081..5128
                     /vntifkey="43"
                     /label=T7\Terminator
     RBS             complement(1045..1049)
                     /vntifkey="32"
                     /label=RBS
     -35_signal      complement(1104..1109)
                     /vntifkey="48"
                     /label=-35\Ap
     -10_signal      complement(1081..1087)
                     /vntifkey="47"
                     /label=-10\Ap
     rep_origin      1149..1784
                     /vntifkey="33"
                     /label=ColA\ori
     -10_signal      complement(3098..3103)
                     /vntifkey="47"
                     /label=lacI\-10
     -35_signal      complement(3122..3127)
                     /vntifkey="48"
                     /label=lacI\-35
     protein_bind    3284..3307
                     /vntifkey="31"
                     /label=Strep-Tag\II
     misc_recomb     3317..3441
                     /vntifkey="86"
                     /label=attR1
     mutation        4065..4065
                     /vntifkey="62"
                     /label=C\to\T\-NcoI
     mutation        4769..4769
                     /vntifkey="62"
                     /label=C\to\T\-BsaI
     CDS             3550..4209
                     /vntifkey="4"
                     /label=Cam
     CDS             4551..4856
                     /vntifkey="4"
                     /label=ccdB
     misc_binding    5023..5040
                     /vntifkey="20"
                     /label=6x\His-Tag
     primer          3274..3313
                     /vntifkey="27"
                     /label=S-NT\Strep\GW
     primer          5017..5052
                     /vntifkey="27"
                     /label=S-CT\6xHis\GW
     misc_recomb     complement(4897..5021)
                     /vntifkey="86"
                     /label=attR2
     misc_feature    3325..5006
                     /vntifkey="21"
                     /label=Region\replaced\by\pENTR\target
     primer          3188..3207
                     /vntifkey="27"
                     /label=T7\Promoter\Primer
BASE COUNT     1364 a      1246 c      1209 g      1315 t 
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 agctggagcc acccgcagtt 
     3301 cgaaaagggt gcagtcacaa gtttgtacaa aaaagctgaa cgagaaacgt aaaatgatat 
     3361 aaatatcaat atattaaatt agattttgca taaaaaacag actacataat actgtaaaac 
     3421 acaacatatc cagtcactat ggcggccgca ttaggcaccc caggctttac actttatgct 
     3481 tccggctcgt ataatgtgtg gattttgagt taggatccgt cgagattttc aggagctaag 
     3541 gaagctaaaa tggagaaaaa aatcactgga tataccaccg ttgatatatc ccaatggcat 
     3601 cgtaaagaac attttgaggc atttcagtca gttgctcaat gtacctataa ccagaccgtt 
     3661 cagctggata ttacggcctt tttaaagacc gtaaagaaaa ataagcacaa gttttatccg 
     3721 gcctttattc acattcttgc ccgcctgatg aatgctcatc cggaattccg tatggcaatg 
     3781 aaagacggtg agctggtgat atgggatagt gttcaccctt gttacaccgt tttccatgag 
     3841 caaactgaaa cgttttcatc gctctggagt gaataccacg acgatttccg gcagtttcta 
     3901 cacatatatt cgcaagatgt ggcgtgttac ggtgaaaacc tggcctattt ccctaaaggg 
     3961 tttattgaga atatgttttt cgtctcagcc aatccctggg tgagtttcac cagttttgat 
     4021 ttaaacgtgg ccaatatgga caacttcttc gcccccgttt tcactatggg caaatattat 
     4081 acgcaaggcg acaaggtgct gatgccgctg gcgattcagg ttcatcatgc cgtttgtgat 
     4141 ggcttccatg tcggcagaat gcttaatgaa ttacaacagt actgcgatga gtggcagggc 
     4201 ggggcgtaaa cgcgtggatc cggcttacta aaagccagat aacagtatgc gtatttgcgc 
     4261 gctgattttt gcggtataag aatatatact gatatgtata cccgaagtat gtcaaaaaga 
     4321 ggtatgctat gaagcagcgt attacagtga cagttgacag cgacagctat cagttgctca 
     4381 aggcatatat gatgtcaata tctccggtct ggtaagcaca accatgcaga atgaagcccg 
     4441 tcgtctgcgt gccgaacgct ggaaagcgga aaatcaggaa gggatggctg aggtcgcccg 
     4501 gtttattgaa atgaacggct cttttgctga cgagaacagg ggctggtgaa atgcagttta 
     4561 aggtttacac ctataaaaga gagagccgtt atcgtctgtt tgtggatgta cagagtgata 
     4621 ttattgacac gcccgggcga cggatggtga tccccctggc cagtgcacgt ctgctgtcag 
     4681 ataaagtctc ccgtgaactt tacccggtgg tgcatatcgg ggatgaaagc tggcgcatga 
     4741 tgaccaccga tatggccagt gtgccggttt ccgttatcgg ggaagaagtg gctgatctca 
     4801 gccaccgcga aaatgacatc aaaaacgcca ttaacctgat gttctgggga atataaatgt 
     4861 caggctccct tatacacagc cagtctgcag gtcgaccata gtgactggat atgttgtgtt 
     4921 ttacagtatt atgtagtctg ttttttatgc aaaatctaat ttaatatatt gatatttata 
     4981 tcattttacg tttctcgttc agctttcttg tacaaagtgg ttcatcacca ccatcatcac 
     5041 taagtgatta acctaggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc 
     5101 ctctaaacgg gtcttgaggg gttttttgct gaaa 
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