LOCUS BGH02_B01a-pET32-DEST 8500 bp DNA circular 15-AUG-2011 FEATURES Location/Qualifiers misc_feature 5208..5534 /note="Trx Tag" misc_feature 5556..5573 /note="His6-tag" misc_feature 8414..8475 /note="T7 Terminator" misc_feature 5139..5163 /note="Lac Operator" misc_feature 5120..5136 /note="T7 Promoter" misc_feature 5535..5552 /note="S Tag" misc_feature 5583..5600 /note="Thrombin" misc_feature 5667..5681 /note="Enterokinase" misc_recomb complement(5693..5721) /label=attB1 /note="attB1 site" misc_recomb 8268..8290 /label=attB2 /note="attB2 site" misc_feature 598..1458 /note="Ap" misc_feature 1613..2232 /note="Ori" misc_feature complement(3647..4606) /note="Lac I" misc_feature 5724..5744 /note="TEV site" misc_feature 5745..8261 /note="BGH02_B01a" vector join(8276..8500,1..5704) /source="pET32-DEST" /type="Destination Vector" clone 5705..8275 /source="BGH02_B01a-pDONR221" /type="Entry Clone" 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 attaacgctt acaatttagg tggcactttt 481 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat 541 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg 601 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt 661 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga 721 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa 781 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt 841 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt 901 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc 961 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga 1021 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat 1081 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct 1141 gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc 1201 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg 1261 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc 1321 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg 1381 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca 1441 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta 1501 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc 1561 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa 1621 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca 1681 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta 1741 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc 1801 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca 1861 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta 1921 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag 1981 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt 2041 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc 2101 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac 2161 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac 2221 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc 2281 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat 2341 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag 2401 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catatatggt 2461 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc 2521 gctacgtgac tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg 2581 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg 2641 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc 2701 atcagcgtgg tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt ccagctcgtt 2761 gagtttctcc agaagcgtta atgtctggct tctgataaag cgggccatgt taagggcggt 2821 tttttcctgt ttggtcactg atgcctccgt gtaaggggga tttctgttca tgggggtaat 2881 gataccgatg aaacgagaga ggatgctcac gatacgggtt actgatgatg aacatgcccg 2941 gttactggaa cgttgtgagg gtaaacaact ggcggtatgg atgcggcggg accagagaaa 3001 aatcactcag ggtcaatgcc agcgcttcgt taatacagat gtaggtgttc cacagggtag 3061 ccagcagcat cctgcgatgc agatccggaa cataatggtg cagggcgctg acttccgcgt 3121 ttccagactt tacgaaacac ggaaaccgaa gaccattcat gttgttgctc aggtcgcaga 3181 cgttttgcag cagcagtcgc ttcacgttcg ctcgcgtatc ggtgattcat tctgctaacc 3241 agtaaggcaa ccccgccagc ctagccgggt cctcaacgac aggagcacga tcatgcgcac 3301 ccgtggggcc gccatgccgg cgataatggc ctgcttctcg ccgaaacgtt tggtggcggg 3361 accagtgacg aaggcttgag cgagggcgtg caagattccg aataccgcaa gcgacaggcc 3421 gatcatcgtc gcgctccagc gaaagcggtc ctcgccgaaa atgacccaga gcgctgccgg 3481 cacctgtcct acgagttgca tgataaagaa gacagtcata agtgcggcga cgatagtcat 3541 gccccgcgcc caccggaagg agctgactgg gttgaaggct ctcaagggca tcggtcgaga 3601 tcccggtgcc taatgagtga gctaacttac attaattgcg ttgcgctcac tgcccgcttt 3661 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg 3721 cggtttgcgt attgggcgcc agggtggttt ttcttttcac cagtgagacg ggcaacagct 3781 gattgccctt caccgcctgg ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc 3841 ccagcaggcg aaaatcctgt ttgatggtgg ttaacggcgg gatataacat gagctgtctt 3901 cggtatcgtc gtatcccact accgagatgt ccgcaccaac gcgcagcccg gactcggtaa 3961 tggcgcgcat tgcgcccagc gccatctgat cgttggcaac cagcatcgca gtgggaacga 4021 tgccctcatt cagcatttgc atggtttgtt gaaaaccgga catggcactc cagtcgcctt 4081 cccgttccgc tatcggctga atttgattgc gagtgagata tttatgccag ccagccagac 4141 gcagacgcgc cgagacagaa cttaatgggc ccgctaacag cgcgatttgc tggtgaccca 4201 atgcgaccag atgctccacg cccagtcgcg taccgtcttc atgggagaaa ataatactgt 4261 tgatgggtgt ctggtcagag acatcaagaa ataacgccgg aacattagtg caggcagctt 4321 ccacagcaat ggcatcctgg tcatccagcg gatagttaat gatcagccca ctgacgcgtt 4381 gcgcgagaag attgtgcacc gccgctttac aggcttcgac gccgcttcgt tctaccatcg 4441 acaccaccac gctggcaccc agttgatcgg cgcgagattt aatcgccgcg acaatttgcg 4501 acggcgcgtg cagggccaga ctggaggtgg caacgccaat cagcaacgac tgtttgcccg 4561 ccagttgttg tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc gcttccactt 4621 tttcccgcgt tttcgcagaa acgtggctgg cctggttcac cacgcgggaa acggtctgat 4681 aagagacacc ggcatactct gcgacatcgt ataacgttac tggtttcaca ttcaccaccc 4741 tgaattgact ctcttccggg cgctatcatg ccataccgcg aaaggttttg cgccattcga 4801 tggtgtccgg gatctcgacg ctctccctta tgcgactcct gcattaggaa gcagcccagt 4861 agtaggttga ggccgttgag caccgccgcc gcaaggaatg gtgcatgcaa ggagatggcg 4921 cccaacagtc ccccggccac ggggcctgcc accataccca cgccgaaaca agcgctcatg 4981 agcccgaagt ggcgagcccg atcttcccca tcggtgatgt cggcgatata ggcgccagca 5041 accgcacctg tggcgccggt gatgccggcc acgatgcgtc cggcgtagag gatcgagatc 5101 gatctcgatc ccgcgaaatt aatacgactc actatagggg aattgtgagc ggataacaat 5161 tcccctctag aaataatttt gtttaacttt aagaaggaga tatacatatg agcgataaaa 5221 ttattcacct gactgacgac agttttgaca cggatgtact caaagcggac ggggcgatcc 5281 tcgtcgattt ctgggcagag tggtgcggtc cgtgcaaaat gatcgccccg attctggatg 5341 aaatcgctga cgaatatcag ggcaaactga ccgttgcaaa actgaacatc gatcaaaacc 5401 ctggcactgc gccgaaatat ggcatccgtg gtatcccgac tctgctgctg ttcaaaaacg 5461 gtgaagtggc ggcaaccaaa gtgggtgcac tgtctaaagg tcagttgaaa gagttcctcg 5521 acgctaacct ggccggttct ggttctggcc atatgcacca tcatcatcat cattcttctg 5581 gtctggtgcc acgcggttct ggtatgaaag aaaccgctgc tgctaaattc gaacgccagc 5641 acatggacag cccagatctg ggtaccgacg acgacgacaa ggccatggcg atATCAACAA 5701 GTTTGTACAA AAAAGCAGGC Tctgaaaact tgtactttca aggcCAAGGT AATGACACTT 5761 CGGAAGTCAT GCTACTGGAT ACCGGTTGGG AATTTTCTCA ATCAGGGACT GAAAAGTGGA 5821 TGCCAGCCAC GGTTCCTGGT ACGGTTCATC AGGACCTGAT CAGTCATGAA TTACTGCCGA 5881 ATCCTTTTTA TGGGATGAAT GAAAAGAAAA TTCAATGGGT GGAGAATGAG GACTGGGAAT 5941 ATAGAACCTC CTTTATAGTC TCCGAAGAGC AATTGAATCG TGACGGAATC CAACTTATTT 6001 TTGAAGGTCT TGATACCTAT GCAGACGTAT ATCTCAACGG CTCATTACTG CTGAAGGCTG 6061 ATAATATGTT TGTGGGTTAT ACACTTCCTG TAAAATCTGT GTTGCGGAAG GGCGAGAATC 6121 ACTTATATAT TTATTTCCAT TCACCTATTC GGCAAACGTT GCCACAATAT GCTTCCAATG 6181 GCTTCAATTA TCCGGCGGAT AACGATCATC ACGAGAAACA TCTTAGTGTG TTCAGCCGTA 6241 AGGCTCCGTA TAGTTACGGT TGGGACTGGG GCATCCGAAT GGTGACAAGT GGTGTCTGGC 6301 GCCCGGTTAC TCTTCGTTTT TACGACATAG CTACCATATC CGACTACTAT GTCAGACAAT 6361 TATCGCTGAC AGATGAAAAC GCTCGTCTGT CTAATGAGCT GATTGTAAAC CAGATCGTTC 6421 CTCAGAAAAT TCCGGCTGAA GTAAGGGTCA ATGTATCGTT GAATGGAACT ACCGTTACAG 6481 AAGTAAAACA GCAGGTGACA CTGCAACCCG GGATAAATCA TATCACTCTT CCTGCTGAAG 6541 TGACGAACCC TGTACGATGG ATGCCAAATG GTTGGGGAAC TCCGACTCTG TATGATTTCT 6601 CGGCTCAGAT AGCTTGTGGA GATCGGATTG TTGCGGAACA ATCTCATCGG ATTGGTTTAC 6661 GAACGATTCG TGTGGTAAAT GAAAAAGATA AGGACGGAGA ATCTTTTTAT TTTGAAGTCA 6721 ATGGCATACC GATGTTTGCC AAAGGAGCTA ATTATATTCC TCAGGATGCA CTGCTGCCTA 6781 ATGTTACTAC GGAACGCTAT CAGACTTTGT TCCGTGACAT GAAGGAGGCC AATATGAATA 6841 TGGTTCGTAT ATGGGGTGGT GGGACATATG AGAATAATCT TTTCTATGAC CTTGCCGATG 6901 AGAACGGGAT ACTTGTCTGG CAGGATTTTA TGTTTGCGTG TACCCCTTAT CCATCCGATC 6961 CTACTTTCCT GAAACGGGTA GAGGCGGAGG CTGTTTACAA TATTCGTCGT CTTCGGAATC 7021 ATGCCTCTTT GGCGATGTGG TGTGGTAATA ATGAAATACT GGAAGCTCTG AAATATTGGG 7081 GTTTTGAGAA GAAGTTTACT CCGGAGGTTT ATCAAGGATT AATGCATGGT TATGATAAAC 7141 TGTTCCGTGA GCTGTTGCCT TCAACGGTTA AAGAATTTGA TTCGGATCGT TTTTATGTGC 7201 ACAGTTCGCC TTATCTCGCT AACTGGGGGC GTCCGGAGTC ATGGGGGACA GGAGACAGTC 7261 ATAATTGGGG AGTCTGGTAC GGAAAGAAGC CATTTGAGTC TTTAGATACC GATTTGCCGC 7321 GTTTTATGAG TGAATTTGGT TTCCAGTCAT TTCCGGAGAT GAAAACTATT GCTGCTTTTG 7381 CTGCTCCTGA AGATTATCAG ATTGAGTCTG AAGTGATGAA TGCGCATCAG AAGAGTAGTA 7441 TTGGTAATTC GTTGATCCGT ACTTATATGG AACGTGATTA TATCATTCCC GAAAGTTTCG 7501 AGGATTTTGT GTATGTCGGA TTAGTCTTGC AAGGACAGGG GATGCGGCAT GGACTGGAAG 7561 CTCACAGACG CAATCGTCCT TATTGTATGG GGACATTGTA TTGGCAGTTG AACGACAGTT 7621 GGCCGGTAGT ATCCTGGTCA AGTATTGATT ATTATGGTAA TTGGAAGGCA TTACATTATC 7681 AGGCAAAGCG TGCTTTTGCA CCGGTCTTGA TCAATCCTAT ACAGCAGAAT GACAGTTTGA 7741 GTGTCTATCT GATCTCAGAC CGGCTTGATA CAATGGAGCA AATGACATTG GAAATGAAAG 7801 TCGTGGATTT TGATGGAAAA ACGCTGGGTA AGAAGATACA AGTACATTCG TTGGAAGTTC 7861 CGGCAAATAC TTCAAAATGT GTTTATCGTG CAAAACTGGA TGGCTGGCTG ACTCCCGAAG 7921 ACTGCCGACG TAGCTTCTTG AAATTGATAC TCAAAGATAA ATCGGGACAT CAGGTGGCTG 7981 AGAGCGTCCA TTTTTTCAGA AAGACGAAAG ATTTGCAACT TCCTCCGACA TCAGTATCCT 8041 ATCAAATGAA GCAGACGGAT GGTAAATGTG AATTGACTCT CTTTTCATCA ATGTTGGCAA 8101 AAGATATTTT TATTGAGACT CCATTGCAAG GAGCCCGTTA CAGTGACAAC TTCTTTGATC 8161 TCTTACCCGG AGAACGTAAG AAAGTAATTA TCACTTCTCC CCGGATAAAG AAGGGAGAAG 8221 AATTACCTGT CAATATCAAG CATATTCGAG AAACCTATAA ATAGGACCCA GCTTTCTTGT 8281 ACAAAGTGGT TGATatcgga tccgaattcg agctccgtcg acaagcttgc ggccgcactc 8341 gagcaccacc accaccacca ctgagatccg gctgctaaca aagcccgaaa ggaagctgag 8401 ttggctgctg ccaccgctga gcaataacta gcataacccc ttggggcctc taaacgggtc 8461 ttgaggggtt ttttgctgaa aggaggaact atatccggat //