LOCUS Exported 10127 bp ds-DNA circular SYN 20-SEP-2016 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS Lsh_C2c2_WT SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 10127) AUTHORS . TITLE Direct Submission JOURNAL Exported Tuesday, Sep 20, 2016 from SnapGene 3.2.1 http://www.snapgene.com FEATURES Location/Qualifiers source 1..10127 /organism="synthetic DNA construct" /mol_type="other DNA" misc_feature complement(139..167) /label=AmpR_promoter /note="AmpR_promoter" misc_feature 209..1069 /label=AMP resistance beta-lactimase /note="AMP resistance beta-lactimase" CDS 407..1066 /label=AmpR /note="AmpR" rep_origin 1164..1846 /label=ColE1 origin /note="ColE1 origin" misc_feature complement(1276..1385) /label=RNA1-for /note="RNA1-for" misc_feature complement(1824..1833) /label=ColE1 ORI /note="ColE1 ORI" misc_feature complement(2257..2482) /label=ROP/ROM /note="ROP/ROM" primer_bind 4010..4029 /label=T7 /note="T7" CDS 4159..4176 /codon_start=1 /product="6xHis affinity tag" /note="6xHis" /translation="HHHHHH" CDS 4186..5286 /codon_start=1 /gene="malE (mutated)" /product="maltose binding protein from E. coli" /note="MBP" /note="This version of the gene does not encode a signal sequence, so MBP will remain in the cytosol." /translation="MKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL ENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAF WYAVRTAVINAASGRQTVDEALKDAQT" CDS 5341..5361 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /note="TEV site" /translation="ENLYFQS" misc_feature 5350..5364 /label=Lic_v1_For /note="Lic_v1_For" misc_feature 5368..9534 /label=WP_018451595.1 /note="Lsh_WT_C2c2" misc_feature 6285..7284 /label=WP_018451595.1-gB2 /note="WP_018451595.1-gB2" misc_feature 7245..8244 /label=WP_018451595.1-gB3 /note="WP_018451595.1-gB3" misc_feature 8205..9581 /label=WP_018451595.1-gB4 /note="WP_018451595.1-gB4" misc_feature 9542..9556 /label=Lic_v1_rev /note="Lic_v1_rev" misc_feature 9663..9681 /label=T7 terminator primer /note="T7 terminator primer" terminator 9677..9724 /note="T7 terminator" /note="transcription terminator for bacteriophage T7 RNA polymerase" misc_feature 9684..9724 /label=T7 terminator /note="T7 terminator" ORIGIN 1 ttcttgaaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat 61 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg 121 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat 181 gcttcaataa cattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat 241 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt 301 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag 361 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa 421 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg 481 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct 541 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac 601 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca 661 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat 721 accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact 781 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc 841 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga 901 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg 961 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg 1021 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca 1081 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta 1141 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca 1201 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg 1261 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga 1321 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa 1381 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc 1441 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg 1501 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac 1561 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct 1621 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc 1681 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg 1741 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg 1801 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct 1861 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga 1921 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg 1981 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca 2041 tctgtgcggt atttcacacc gcatatatgg tgcactctca gtacaatctg ctctgatgcc 2101 gcatagttaa gccagtatac actccgctat cgctacgtga ctgggtcatg gctgcgcccc 2161 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt 2221 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac 2281 cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc gattcacaga 2341 tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt aatgtctggc 2401 ttctgataaa gcgggccatg ttaagggcgg ttttttcctg tttggtcact gatgcctccg 2461 tgtaaggggg atttctgttc atgggggtaa tgataccgat gaaacgagag aggatgctca 2521 cgatacgggt tactgatgat gaacatgccc ggttactgga acgttgtgag ggtaaacaac 2581 tggcggtatg gatgcggcgg gaccagagaa aaatcactca gggtcaatgc cagcgcttcg 2641 ttaatacaga tgtaggtgtt ccacagggta gccagcagca tcctgcgatg cagatccgga 2701 acataatggt gcagggcgct gacttccgcg tttccagact ttacgaaaca cggaaaccga 2761 agaccattca tgttgttgct caggtcgcag acgttttgca gcagcagtcg cttcacgttc 2821 gctcgcgtat cggtgattca ttctgctaac cagtaaggca accccgccag cctagccggg 2881 tcctcaacga caggagcacg atcatgcgca cccgtggcca ggacccaacg ctgcccgaga 2941 tgcgccgcgt gcggctgctg gagatggcgg acgcgatgga tatgttctgc caagggttgg 3001 tttgcgcatt cacagttctc cgcaagaatt gattggctcc aattcttgga gtggtgaatc 3061 cgttagcgag gtgccgccgg cttccattca ggtcgaggtg gcccggctcc atgcaccgcg 3121 acgcaacgcg gggaggcaga caaggtatag ggcggcgcct acaatccatg ccaacccgtt 3181 ccatgtgctc gccgaggcgg cataaatcgc cgtgacgatc agcggtccag tgatcgaagt 3241 taggctggta agagccgcga gcgatccttg aagctgtccc tgatggtcgt catctacctg 3301 cctggacagc atggcctgca acgcgggcat cccgatgccg ccggaagcga gaagaatcat 3361 aatggggaag gccatccagc ctcgcgtcgc gaacgccagc aagacgtagc ccagcgcgtc 3421 ggccgccatg ccggcgataa tggcctgctt ctcgccgaaa cgtttggtgg cgggaccagt 3481 gacgaaggct tgagcgaggg cgtgcaagat tccgaatacc gcaagcgaca ggccgatcat 3541 cgtcgcgctc cagcgaaagc ggtcctcgcc gaaaatgacc cagagcgctg ccggcacctg 3601 tcctacgagt tgcatgataa agaagacagt cataagtgcg gcgacgatag tcatgccccg 3661 cgcccaccgg aaggagctga ctgggttgaa ggctctcaag ggcatcggtc gacgctctcc 3721 cttatgcgac tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt tgagcaccgc 3781 cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac agtcccccgg ccacggggcc 3841 tgccaccata cccacgccga aacaagcgct catgagcccg aagtggcgag cccgatcttc 3901 cccatcggtg atgtcggcga tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc 3961 ggccacgatg cgtccggcgt agaggatcga gatctcgatc ccgcgaaatt aatacgactc 4021 actataggga gaccacaacg gtttccctct agtgccggct ccggagagct ctttaattaa 4081 gcggccgccc tgcaggactc gagttctaga aataattttg tttaacttta agaaggagat 4141 atacatatga aatcttctca ccatcaccat caccatggtt cttctatgaa aatcgaagaa 4201 ggtaaactgg taatctggat taacggcgat aaaggctata acggtctcgc tgaagtcggt 4261 aagaaattcg agaaagatac cggaattaaa gtcaccgttg agcatccgga taaactggaa 4321 gagaaattcc cacaggttgc ggcaactggc gatggccctg acattatctt ctgggcacac 4381 gaccgctttg gtggctacgc tcaatctggc ctgttggctg aaatcacccc ggacaaagcg 4441 ttccaggaca agctgtatcc gtttacctgg gatgccgtac gttacaacgg caagctgatt 4501 gcttacccga tcgctgttga agcgttatcg ctgatttata acaaagatct gctgccgaac 4561 ccgccaaaaa cctgggaaga gatcccggcg ctggataaag aactgaaagc gaaaggtaag 4621 agcgcgctga tgttcaacct gcaagaaccg tacttcacct ggccgctgat tgctgctgac 4681 gggggttatg cgttcaagta tgaaaacggc aagtacgaca ttaaagacgt gggcgtggat 4741 aacgctggcg cgaaagcggg tctgaccttc ctggttgacc tgattaaaaa caaacacatg 4801 aatgcagaca ccgattactc catcgcagaa gctgccttta ataaaggcga aacagcgatg 4861 accatcaacg gcccgtgggc atggtccaac atcgacacca gcaaagtgaa ttatggtgta 4921 acggtactgc cgaccttcaa gggtcaacca tccaaaccgt tcgttggcgt gctgagcgca 4981 ggtattaacg ccgccagtcc gaacaaagag ctggcaaaag agttcctcga aaactatctg 5041 ctgactgatg aaggtctgga agcggttaat aaagacaaac cgctgggtgc cgtagcgctg 5101 aagtcttacg aggaagagtt ggcgaaagat ccacgtattg ccgccactat ggaaaacgcc 5161 cagaaaggtg aaatcatgcc gaacatcccg cagatgtccg ctttctggta tgccgtgcgt 5221 actgcggtga tcaacgccgc cagcggtcgt cagactgtcg atgaagccct gaaagacgcg 5281 cagactaatt cgagctcgaa caacaacaac aataacaata acaacaacct cgggatcgag 5341 gaaaacctgt acttccaatc caatgcaatg gggaaccttt tcgggcataa acgctggtac 5401 gaggtgcgcg ataagaagga ctttaagatc aaacgcaaag tcaaggttaa gcgtaattac 5461 gacggtaaca aatacatctt aaacatcaac gagaacaaca ataaggagaa aatcgacaat 5521 aacaagttta ttcgtaagta catcaattac aagaaaaacg acaatatcct taaagaattt 5581 acgcgcaaat tccatgcagg caatatcttg ttcaagttaa agggtaagga ggggatcatt 5641 cgtatcgaga acaatgatga ctttctggaa actgaagagg ttgtccttta catcgaagcg 5701 tatgggaaga gcgagaagct gaaagctctg ggaatcacta aaaaaaaaat cattgacgag 5761 gcgattcgtc aaggcatcac aaaggatgac aagaaaatcg aaatcaagcg ccaagagaac 5821 gaagaagaaa tcgaaatcga catccgcgat gaatacacca acaagacctt aaatgattgc 5881 agtattattc tgcgcatcat tgaaaacgat gagcttgaga cgaaaaagtc tatctatgag 5941 atctttaaga atatcaacat gtcgttgtac aagattatcg agaagattat tgaaaatgag 6001 actgagaaag tctttgaaaa ccgttattat gaagagcacc ttcgcgaaaa actgcttaag 6061 gacgacaaaa ttgatgttat tctgactaac tttatggaga ttcgtgagaa gatcaagagc 6121 aacctggaaa ttttagggtt cgtcaaattc tacttaaacg taggggggga taaaaagaag 6181 tccaaaaaca agaagatgtt ggtggagaaa atcttaaaca tcaatgtaga cctgaccgtg 6241 gaggacattg ctgattttgt tattaaggag ttggagtttt ggaacatcac aaaacgcatt 6301 gaaaaagtta agaaagttaa taacgagttt ttagaaaagc gtcgcaaccg cacgtatatc 6361 aaatcatacg tccttttaga taagcacgaa aaatttaaga tcgagcgcga aaacaagaaa 6421 gataaaattg tcaaattttt cgtcgagaat atcaagaaca atagcatcaa ggagaagatc 6481 gaaaagatct tggcggaatt taaaatcgac gaattgatca aaaaattaga gaaagagtta 6541 aagaaaggta actgcgatac agagatcttc ggtatcttca agaaacacta taaggtcaat 6601 tttgactcca agaagttctc caagaaatct gacgaggaaa aggaattgta taagattatc 6661 tatcgctacc ttaaaggacg catcgagaag attctggtca acgaacagaa ggtacgctta 6721 aagaagatgg agaaaatcga aattgagaag attttaaacg aatctatcct ttcggaaaaa 6781 atccttaagc gcgtcaagca gtacaccctg gaacacatca tgtacttggg aaaacttcgt 6841 cataatgata ttgatatgac gactgtcaat actgatgatt tttctcgctt acatgcgaaa 6901 gaagagctgg acttggagtt gattacgttt ttcgcatcta cgaacatgga actgaacaag 6961 attttctctc gcgagaatat taataacgac gaaaatattg acttctttgg aggtgaccgc 7021 gaaaagaact atgtgctcga caagaaaatt ctcaactcta agattaagat cattcgcgat 7081 cttgatttca tcgacaataa gaacaacatt acgaataact ttattcgcaa gtttaccaag 7141 attggtacga atgagcgcaa tcgtattttg cacgccatta gtaaagaacg tgatttgcag 7201 ggaacgcaag acgactataa caaggtaatc aacatcattc aaaacttgaa gatctccgac 7261 gaagaagtat caaaagcctt gaatttggat gtcgtgttca aggataagaa gaacattatc 7321 acaaaaatca acgatattaa aatcagtgag gagaataaca acgatattaa atatttaccg 7381 agttttagca aggttctgcc ggaaatctta aatttatacc gcaataatcc taaaaacgaa 7441 cccttcgata caattgagac tgaaaagatc gtgttgaacg ctcttattta tgtgaacaaa 7501 gaattgtaca agaagttaat cctggaggat gacctggaag agaacgagag caaaaacatt 7561 ttcttgcagg agctgaagaa aaccttaggt aatattgatg agatcgacga gaacatcatt 7621 gagaattact acaagaacgc ccaaatctcg gcgtctaaag gaaataataa ggcgattaag 7681 aaataccaga agaaagttat cgaatgctac atcgggtatt tgcgtaagaa ctacgaagag 7741 cttttcgact tttcggactt taagatgaac atccaagaga ttaagaaaca aatcaaggat 7801 attaatgaca ataagacgta tgagcgtatt acagtgaaaa ccagtgacaa gactatcgta 7861 attaacgacg attttgagta catcatttca atctttgcgt tgttgaactc taatgccgta 7921 atcaacaaga ttcgcaaccg tttcttcgcg acgtcagtgt ggctgaacac ctcagagtac 7981 cagaatatta tcgatatcct ggacgagatt atgcagctga atacgctgcg taacgagtgt 8041 attaccgaaa actggaacct caacttagaa gagtttatcc aaaagatgaa ggaaatcgag 8101 aaggactttg atgattttaa aattcagaca aagaaggaga tctttaacaa ctattacgag 8161 gacatcaaaa acaacatctt aacggaattt aaagacgata ttaacggttg cgacgtgctt 8221 gaaaagaagt tggagaaaat cgtaatcttt gacgacgaaa ccaagtttga gatcgacaag 8281 aagtccaata ttttgcagga cgagcagcgt aaattgagca atatcaataa gaaggacctg 8341 aaaaagaagg tggaccaata tattaaagac aaggatcagg aaatcaaatc gaagattttg 8401 tgccgtatta tcttcaactc agacttcctt aagaaatata agaaggaaat tgacaatttg 8461 atcgaagata tggagtctga gaatgagaac aagttccaag agatttatta cccgaaagag 8521 cgcaagaacg aactctatat ctataagaaa aacttgttct taaatattgg aaacccaaac 8581 tttgacaaga tttatggcct tatttcgaac gatattaaga tggccgatgc taaattcttg 8641 tttaatatcg atggtaagaa tattcgtaaa aataagatta gtgaaattga cgccattttg 8701 aagaacctta acgacaagct gaacggctac tcgaaagagt acaaggagaa gtatatcaaa 8761 aagttgaagg aaaacgatga cttctttgcc aaaaatatcc agaacaaaaa ttacaagagc 8821 ttcgaaaagg attacaaccg cgtttctgag tataaaaaga tccgcgacct tgtagagttc 8881 aactatttaa acaaaatcga atcttactta atcgatatca actggaaatt ggcgattcaa 8941 atggcccgct ttgagcgcga catgcactat atcgtaaatg ggttgcgcga gttgggaatt 9001 atcaagttgt caggatataa cacggggatc tcacgtgcat acccaaagcg caatggatca 9061 gacgggtttt acacaaccac tgcttactat aagttctttg atgaggaatc ctacaagaaa 9121 ttcgaaaaga tctgttacgg tttcggtatc gacttgagcg aaaacagcga gatcaacaag 9181 ccagaaaatg aatccattcg caactatatt tctcactttt atatcgttcg caacccgttt 9241 gcagactact ctattgcgga gcagatcgac cgtgtgtcga atttgctgag ttatagcacc 9301 cgctataaca actcgacgta tgcatcagta ttcgaggtct ttaaaaagga cgtgaacctt 9361 gactatgatg aattaaagaa aaaattcaag cttattggca ataatgacat cttggaacgc 9421 ttgatgaagc cgaagaaagt ctccgtactt gaattagaat cgtacaactc tgattacatc 9481 aagaacctga tcattgagtt attgactaag attgaaaata caaacgatac cctgtaataa 9541 cattggaagt ggataacgga tccgcgatcg cggcgcgcca cctggtggcc ggccggtacc 9601 acgcgtgcgc gctgatccgg ctgctaacaa agcccgaaag gaagctgagt tggctgctgc 9661 caccgctgag caataactag cataacccct tggggcctct aaacgggtct tgaggggttt 9721 tttgctgaaa ggaggaacta tatccggata tccacaggac gggtgtggtc gccatgatcg 9781 cgtagtcgat agtggctcca agtagcgaag cgagcaggac tgggcggcgg ccaaagcggt 9841 cggacagtgc tccgagaacg ggtgcgcata gaaattgcat caacgcatat agcgctagca 9901 gcacgccata gtgactggcg atgctgtcgg aatggacgat atcccgcaag aggcccggca 9961 gtaccggcat aaccaagcct atgcctacag catccagggt gacggtgccg aggatgacga 10021 tgagcgcatt gttagatttc atacacggtg cctgactgcg ttagcaattt aactgtgata 10081 aactaccgca ttaaagctta tcgatgataa gctgtcaaac atgagaa //