LOCUS Exported 6279 bp ds-DNA circular SYN 13-PAŹ-2016 DEFINITION synthetic circular DNA. ACCESSION . VERSION . KEYWORDS pKK-TEV-MBP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6279) AUTHORS . TITLE Direct Submission JOURNAL Exported 14 paza 2016 from SnapGene 3.0.3 http://www.snapgene.com COMMENT LSOWNER| FEATURES Location/Qualifiers source 1..6279 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 232..958 /note="CMV promoter" /note="human cytomegalovirus (CMV) immediate early promoter" protein_bind 820..838 /bound_moiety="repressor" /note="tet operator" protein_bind 841..859 /bound_moiety="repressor" /note="tet operator" misc_feature 1002..1022 /note="TEV-L" /note="enocdes peptide cleaved by TEV protease" misc_feature 1063..1083 /note="TEV-R" /note="enocdes peptide cleaved by TEV protease" CDS 1090..2190 /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="MKTEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL ENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAF WYAVRTAVINAASGRQTVDEALKDAQT" polyA_signal 2237..2461 /note="BGH poly(A)signal" /note="bovine growth hormone polyadenylation signal" protein_bind 2745..2792 /bound_moiety="FLP recombinase" /note="FRT" CDS 2800..3822 /codon_start=1 /product="aminoglycoside phosphotransferase from E. coli" /note="Hygromycin B" /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE " polyA_signal 3952..4082 /note="SV40 poly(A)signal" /note="SV40 polyadenylation signal" rep_origin complement(4465..5138) /direction=LEFT /note="pUC ori" CDS complement(5283..6143) /codon_start=1 /product="beta-lactamase" /note="AmpR" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" misc_feature complement(6144..6242) /note="Amp promoter" ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg 61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt 241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata 301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc 361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc 421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt 481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt 541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca 601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg 661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc 721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg 781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ccctatcagt gatagagatc 841 tccctatcag tgatagagat cgtcgacgag ctcgtttagt gaaccgtcag atcgcctgga 901 gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc agcctccgga 961 ctctagcgtt taaacttaag cttggtaccg agctcggatc cgaaaacctg tacttccaag 1021 gaaccggtgg tcacctgtac aatcgatcct gcaggagcta gcgagaattt gtattttcag 1081 ggtgatatca tgaaaactga agaaggtaaa ctggtaatct ggattaacgg cgataaaggc 1141 tataacggtc tcgctgaagt cggtaagaaa ttcgagaaag ataccggaat taaagtcacc 1201 gttgagcatc cggataaact ggaagagaaa ttcccacagg ttgcggcaac tggcgatggc 1261 cctgacatta tcttctgggc acacgaccgc tttggtggct acgctcaatc tggcctgttg 1321 gctgaaatca ccccggacaa agcgttccag gacaagctgt atccgtttac ctgggatgcc 1381 gtacgttaca acggcaagct gattgcttac ccgatcgctg ttgaagcgtt atcgctgatt 1441 tataacaaag atctgctgcc gaacccgcca aaaacctggg aagagatccc ggcgctggat 1501 aaagaactga aagcgaaagg taagagcgcg ctgatgttca acctgcaaga accgtacttc 1561 acctggccgc tgattgctgc tgacgggggt tatgcgttca agtatgaaaa cggcaagtac 1621 gacattaaag acgtgggcgt ggataacgct ggcgcgaaag cgggtctgac cttcctggtt 1681 gacctgatta aaaacaaaca catgaatgca gacaccgatt actccatcgc agaagctgcc 1741 tttaataaag gcgaaacagc gatgaccatc aacggcccgt gggcatggtc caacatcgac 1801 accagcaaag tgaattatgg tgtaacggta ctgccgacct tcaagggtca accatccaaa 1861 ccgttcgttg gcgtgctgag cgcaggtatt aacgccgcca gtccgaacaa agagctggcg 1921 aaagagttcc tcgaaaacta tctgctgact gatgaaggtc tggaagcggt taataaagac 1981 aaaccgctgg gtgccgtagc gctgaagtct tacgaggaag agttggcgaa agatccacgt 2041 attgccgcca ccatggaaaa cgcccagaaa ggtgaaatca tgccgaacat cccgcagatg 2101 tccgctttct ggtatgccgt gcgtactgcg gtgatcaacg ccgccagcgg tcgtcagact 2161 gtcgatgaag ccctgaaaga cgcgcagact taactcgagt ctagagggcc cgtttaaacc 2221 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc 2281 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa 2341 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac 2401 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg 2461 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc 2521 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc 2581 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt 2641 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac 2701 ctcgacccca aaaaacttga ttagggtgat ggttcacgta cctagaagtt cctattccga 2761 agttcctatt ctctagaaag tataggaact tccttggcca aaaagcctga actcaccgcg 2821 acgtctgtcg agaagtttct gatcgaaaag ttcgacagcg tctccgacct gatgcagctc 2881 tcggagggcg aagaatctcg tgctttcagc ttcgatgtag gagggcgtgg atatgtcctg 2941 cgggtaaata gctgcgccga tggtttctac aaagatcgtt atgtttatcg gcactttgca 3001 tcggccgcgc tcccgattcc ggaagtgctt gacattgggg aattcagcga gagcctgacc 3061 tattgcatct cccgccgtgc acagggtgtc acgttgcaag acctgcctga aaccgaactg 3121 cccgctgttc tgcagccggt cgcggaggcc atggatgcga tcgctgcggc cgatcttagc 3181 cagacgagcg ggttcggccc attcggaccg caaggaatcg gtcaatacac tacatggcgt 3241 gatttcatat gcgcgattgc tgatccccat gtgtatcact ggcaaactgt gatggacgac 3301 accgtcagtg cgtccgtcgc gcaggctctc gatgagctga tgctttgggc cgaggactgc 3361 cccgaagtcc ggcacctcgt gcacgcggat ttcggctcca acaatgtcct gacggacaat 3421 ggccgcataa cagcggtcat tgactggagc gaggcgatgt tcggggattc ccaatacgag 3481 gtcgccaaca tcttcttctg gaggccgtgg ttggcttgta tggagcagca gacgcgctac 3541 ttcgagcgga ggcatccgga gcttgcagga tcgccgcggc tccgggcgta tatgctccgc 3601 attggtcttg accaactcta tcagagcttg gttgacggca atttcgatga tgcagcttgg 3661 gcgcagggtc gatgcgacgc aatcgtccga tccggagccg ggactgtcgg gcgtacacaa 3721 atcgcccgca gaagcgcggc cgtctggacc gatggctgtg tagaagtact cgccgatagt 3781 ggaaaccgac gccccagcac tcgtccgagg gcaaaggaat agcacgtact acgagatttc 3841 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc 3901 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt 3961 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca 4021 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc 4081 tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg 4141 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa 4201 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct 4261 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga 4321 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc 4381 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa 4441 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt 4501 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa 4561 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt 4621 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg 4681 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc 4741 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc 4801 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta 4861 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct 4921 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc 4981 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa 5041 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa 5101 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa 5161 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt 5221 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac 5281 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc 5341 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc 5401 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata 5461 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc 5521 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc 5581 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca 5641 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa 5701 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca 5761 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt 5821 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt 5881 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg 5941 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga 6001 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc 6061 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg 6121 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag 6181 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg 6241 gttccgcgca catttccccg aaaagtgcca cctgacgtc //