LOCUS Exported 9437 bp ds-DNA circular SYN 13-APR-2017 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS p2CT-His-MBP-Lbu_C2c2_E299A SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9437) AUTHORS . TITLE Direct Submission JOURNAL Exported Thursday, Apr 13, 2017 from SnapGene 3.3.3 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9437 /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 4147..5349 /codon_start=1 /label=MBP ORF /note="MBP ORF" /translation="MKSSHHHHHHGSSMKIEEGKLVIWINGDKGYNGLAEVGKKFEKDT GIKVTVEHPDKLEEKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLY PFTWDAVRYNGKLIAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMF NLQEPYFTWPLIAADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADT DYSIAEAAFNKGETAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGIN AASPNKELAKEFLENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQK GEIMPNIPQMSAFWYAVRTAVINAASGRQTVDEALKDAQTNSSSNNNNNNNNNNLGIEE NL" misc_feature 4159..4176 /label=His6 tag /note="His6 tag" misc_feature 5325..6574 /label=WP_015770004.1_gB1 /note="WP_015770004.1_gB1" 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" CDS 5368..8844 /codon_start=1 /label=WP_015770004.1 /note="Lbu_C2c2_WT" /translation="MKVTKVGGISHKKYTSEGRLVKSESEENRTDERLSALLNMRLDMY IKNPSSTETKENQKRIGKLKKFFSNKMVYLKDNTLSLKNGKKENIDREYSETDILESDV RDKKNFAVLKKIYLNENVNSEELEVFRNDIKKKLNKINSLKYSFEKNKANYQKINENNI EKVEGKSKRNIIYDYYRESAKRDAYVSNVKEAFDKLYKEEDIAKLVLEIENLTKLEKYK IREFYHEIIGRKNDKENFAKIIYEEIQNVNNMKELIEKVPDMSELKKSQVFYKYYLDKE ELNDKNIKYAFCHFVEIAMSQLLKNYVYKRLSNISNDKIKRIFEYQNLKKLIENKLLNK LDTYVRNCGKYNYYLQDGEIATSDFIARNRQNEAFLRNIIGVSSVAYFSLRNILETENE NDITGRMRGKTVKNNKGEEKYVSGEVDKIYNENKKNEVKENLKMFYSYDFNMDNKNEIE DFFANIDEAISSIRHGIVHFNLELEGKDIFAFKNIAPSEISKKMFQNEINEKKLKLKIF RQLNSANVFRYLEKYKILNYLKRTRFEFVNKNIPFVPSFTKLYSRIDDLKNSLGIYWKT PKTNDDNKTKEIIDAQIYLLKNIYYGEFLNYFMSNNGNFFEISKEIIELNKNDKRNLKT GFYKLQKFEDIQEKIPKEYLANIQSLYMINAGNQDEEEKDTYIDFIQKIFLKGFMTYLA NNGRLSLIYIGSDEETNTSLAEKKQEFDKFLKKYEQNNNIKIPYEINEFLREIKLGNIL KYTERLNMFYLILKLLNHKELTNLKGSLEKYQSANKEEAFSDQLELINLLNLDNNRVTE DFELEADEIGKFLDFNGNKVKDNKELKKFDTNKIYFDGENIIKHRAFYNIKKYGMLNLL EKIADKAGYKISIEELKKYSNKKNEIEKNHKMQENLHRKYARPRKDEKFTDEDYESYKQ AIENIEEYTHLKNKVEFNELNLLQGLLLRILHRLVGYTSIWERDLRFRLKGEFPENQYI EEIFNFENKKNVKYKGGQIVEKYIKFYKELHQNDEVKINKYSSANIKVLKQEKKDLYIR NYIAHFNYIPHAEISLLEVLENLRKLLSYDRKLKNAVMKSVVDILKEYGFVATFKIGAD KKIGIQTLESEKIVHLKNLKKKKLMTDRNSEELCKLVKIMFEYKMEEKKSEN" CDS 6244..6246 /codon_start=1 /note="Cystine" /translation="C" CDS 6409..6411 /codon_start=1 /note="Cystine" /translation="C" misc_feature 6535..7784 /label=WP_015770004.1_gB2 /note="WP_015770004.1_gB2" misc_feature 6783..6795 /note="Active Site 1" misc_feature 7745..8891 /label=WP_015770004.1_gB3 /note="WP_015770004.1_gB3" misc_feature 7960..8307 /note="Phyre aligned section" misc_feature 8512..8529 /note="Active Site 2" CDS 8530..8847 /codon_start=1 /note="tail" /translation="NYIPHAEISLLEVLENLRKLLSYDRKLKNAVMKSVVDILKEYGFV ATFKIGADKKIGIQTLESEKIVHLKNLKKKKLMTDRNSEELCKLVKIMFEYKMEEKKSE N" CDS 8788..8790 /codon_start=1 /note="cysteine" /translation="C" misc_feature 8852..8866 /label=Lic_v1_rev /note="Lic_v1_rev" misc_feature 8973..8991 /label=T7 terminator primer /note="T7 terminator primer" misc_feature 8994..9034 /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 aaagtgacga aggtaggagg catttcgcat 5401 aagaagtaca cgtccgaagg ccgcttagtg aagtcagaat cggaagaaaa tcgcacagac 5461 gaacgtctgt cggcgttgct taatatgcgc cttgacatgt atatcaagaa tcccagcagc 5521 acggaaacca aggaaaatca aaaacgcatt gggaaattaa agaaattctt ctcaaacaaa 5581 atggtctatc ttaaagacaa taccttgagt ttgaagaatg ggaaaaagga gaacattgat 5641 cgtgagtatt ctgagactga catccttgag agcgatgtcc gtgacaagaa aaacttcgcc 5701 gtgttgaaaa agatctatct gaatgaaaac gtgaactcgg aggaattgga agtttttcgt 5761 aacgacatta agaagaaact gaacaaaatc aacagcctga agtactcatt tgaaaagaat 5821 aaggcgaatt atcaaaagat taatgagaat aacatcgaga aggttgaagg taagtcaaag 5881 cgtaacatta tttacgatta ttatcgtgag tcagcgaaac gtgacgctta tgtaagcaat 5941 gtgaaagaag cctttgataa gctttacaag gaagaggaca ttgcaaaact tgttcttgaa 6001 attgagaacc ttacgaagtt agagaaatac aagattcgcg agttctacca cgaaattatt 6061 ggacgtaaga atgacaagga aaactttgca aaaatcatct acgaagaaat ccagaatgtt 6121 aataacatga aagagttgat cgagaaggta ccggacatga gtgagttgaa aaagagccaa 6181 gtattttaca agtattactt agacaaagaa gagttgaacg acaagaacat caaatacgcg 6241 ttttgtcatt tcgtggaaat cgccatgagt cagttgctga agaactacgt atataagcgc 6301 ttaagtaata tctcgaatga caaaattaag cgtatctttg aataccagaa cttgaaaaaa 6361 ttgatcgaaa ataagctgtt aaacaaactt gacacgtacg tccgtaattg tggaaagtat 6421 aattattatt tgcaagacgg cgaaattgcc acttcagatt tcatcgcccg caaccgtcag 6481 aatgaagcgt ttcttcgcaa catcattggg gtgtcatctg tggcctactt ttctcttcgc 6541 aacattcttg aaacggagaa cgagaatgat attactgggc gtatgcgcgg caaaacagtt 6601 aagaacaata aaggtgaaga gaagtacgtg tccggagaag ttgataagat ctataatgaa 6661 aataagaaga acgaggttaa ggagaactta aaaatgttct attcgtacga tttcaatatg 6721 gacaacaaga atgaaatcga agatttcttc gccaacatcg acgaggcgat ttcttccatc 6781 cgtcacggta ttgtccactt caacttggaa ttagaaggta aggatatctt tgcgttcaag 6841 aacattgcgc catccgaaat ctcaaagaag atgtttcaga atgagattaa cgagaaaaaa 6901 ctgaaattga agatctttcg tcaactgaac tctgccaacg tgttccgcta tctcgaaaag 6961 tataaaattc tgaattacct taaacgtaca cgcttcgagt ttgtcaataa aaatatccca 7021 ttcgtcccgt ctttcaccaa attatattcg cgcattgatg acctgaagaa tagtcttggg 7081 atttactgga aaactccgaa aacaaacgac gacaataaga ctaaggagat tattgatgcc 7141 caaatctatt tgcttaaaaa catctattac ggggagttcc tgaattattt catgtcgaac 7201 aatggtaatt tctttgagat ttctaaagaa atcatcgaat tgaacaagaa cgataaacgc 7261 aacttaaaga ctgggtttta caagctgcaa aagtttgaag acatccagga gaagattcca 7321 aaggaatact tggcgaatat ccagtccctg tacatgatta atgccggtaa tcaggacgaa 7381 gaagaaaagg acacttatat tgatttcatt caaaagatct tcttaaaggg atttatgacg 7441 tatcttgcta ataacggtcg tttaagtctg atttacatcg gctcggatga agaaacaaat 7501 acgtcattag cagaaaagaa gcaagagttt gacaagttct tgaagaagta cgagcagaac 7561 aataatatca agatccccta tgagatcaat gaattcctgc gtgagatcaa actgggaaac 7621 atcctgaagt atactgagcg tttaaacatg ttctacctta tcttaaagct tttgaatcac 7681 aaggagctga caaatctgaa gggtagtctt gaaaaatatc agtctgccaa taaggaagaa 7741 gcgttctctg accaattgga gttaattaac ctgcttaacc ttgacaacaa ccgcgtgacg 7801 gaagacttcg aattagaggc cgacgagatt ggaaaatttc ttgatttcaa tggcaacaaa 7861 gttaaggata acaaggaact gaaaaagttc gatacaaaca agatctactt tgacggcgag 7921 aacattatca aacaccgtgc cttctacaat attaagaaat atggcatgtt aaacttactg 7981 gagaaaattg ccgacaaggc tggatacaag atctcgatcg aagagctgaa gaaatactcc 8041 aataaaaaga atgagatcga gaagaaccat aagatgcagg aaaatctgca ccgcaaatac 8101 gctcgtcccc gtaaagacga gaagtttaca gatgaggact atgaaagtta caagcaagct 8161 attgagaata ttgaggagta cacccacctt aagaacaagg tagaattcaa tgagctgaat 8221 ttactgcagg gcctgttgct gcgcatttta catcgtttag tcggatatac ctcaatttgg 8281 gaacgcgatc tgcgcttccg ccttaaaggt gagttcccag aaaaccaata catcgaagag 8341 atcttcaact ttgaaaataa gaagaacgtg aagtacaaag ggggtcagat tgtagagaaa 8401 tacattaaat tctacaagga attacatcaa aatgatgaag ttaagatcaa caagtacagt 8461 tccgcgaata tcaaggtgtt gaagcaagaa aagaaggacc tttatattcg aaattacatc 8521 gcccacttca attatattcc tcacgccgag atctcactgc tggaagtcct tgaaaatttg 8581 cgtaaattgc tgtcctacga tcgcaaactg aaaaatgccg taatgaaatc agtagttgat 8641 atccttaagg agtatggttt tgtagccaca ttcaaaatcg gggcggacaa gaagatcggt 8701 attcagacac tggagagcga aaaaatcgtg catcttaaga atcttaagaa gaagaagtta 8761 atgactgacc gcaattccga ggaactttgc aaattggtga agattatgtt tgaatacaaa 8821 atggaagaga aaaagtctga aaactaataa cattggaagt ggataacgga tccgcgatcg 8881 cggcgcgcca cctggtggcc ggccggtacc acgcgtgcgc gctgatccgg ctgctaacaa 8941 agcccgaaag gaagctgagt tggctgctgc caccgctgag caataactag cataacccct 9001 tggggcctct aaacgggtct tgaggggttt tttgctgaaa ggaggaacta tatccggata 9061 tccacaggac gggtgtggtc gccatgatcg cgtagtcgat agtggctcca agtagcgaag 9121 cgagcaggac tgggcggcgg ccaaagcggt cggacagtgc tccgagaacg ggtgcgcata 9181 gaaattgcat caacgcatat agcgctagca gcacgccata gtgactggcg atgctgtcgg 9241 aatggacgat atcccgcaag aggcccggca gtaccggcat aaccaagcct atgcctacag 9301 catccagggt gacggtgccg aggatgacga tgagcgcatt gttagatttc atacacggtg 9361 cctgactgcg ttagcaattt aactgtgata aactaccgca ttaaagctta tcgatgataa 9421 gctgtcaaac atgagaa //