LOCUS Exported 5877 bp ds-DNA circular SYN 24-MAY-2017 DEFINITION synthetic circular DNA. ACCESSION . VERSION . KEYWORDS pKK-TEV-mTagBFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5877) AUTHORS . TITLE Direct Submission JOURNAL Exported Wednesday, May 24, 2017 from SnapGene 3.3.3 http://www.snapgene.com COMMENT LSOWNER| FEATURES Location/Qualifiers source 1..5877 /organism="synthetic DNA construct" /mol_type="other DNA" source 1090..1788 /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..1788 /codon_start=1 /product="monomeric blue fluorescent protein" /note="mTagBFP" /note="mammalian codon-optimized" /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC DLPSKLGHKLN" polyA_signal 1835..2059 /note="BGH poly(A)signal" /note="bovine growth hormone polyadenylation signal" protein_bind 2343..2390 /bound_moiety="FLP recombinase" /note="FRT" CDS 2398..3420 /codon_start=1 /product="aminoglycoside phosphotransferase from E. coli" /note="Hygromycin B" /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE " polyA_signal 3550..3680 /note="SV40 poly(A)signal" /note="SV40 polyadenylation signal" rep_origin complement(4063..4736) /direction=LEFT /note="pUC ori" CDS complement(4881..5741) /codon_start=1 /product="beta-lactamase" /note="AmpR" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" misc_feature complement(5742..5840) /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 ctctggcgtt taaacttaag cttggtaccg agctcggatc cgaaaacctg tacttccaag 1021 gaaccggtgg tcacctgtac aatcgatcct gcaggagcta gcgagaattt gtattttcag 1081 ggtgatatca tgagcgagct gattaaggag aacatgcaca tgaagctgta catggagggc 1141 accgtggaca accatcactt caagtgcaca tccgagggcg aaggcaagcc ctacgagggc 1201 acccagacca tgagaatcaa ggtggtcgag ggcggccctc tccccttcgc cttcgacatc 1261 ctggctacta gcttcctcta cggcagcaag accttcatca accacaccca gggcatcccc 1321 gacttcttca agcagtcctt ccctgagggc ttcacatggg agagagtcac cacatacgaa 1381 gacgggggcg tgctgaccgc tacccaggac accagcctcc aggacggctg cctcatctac 1441 aacgtcaaga tcagaggggt gaacttcaca tccaacggcc ctgtgatgca gaagaaaaca 1501 ctcggctggg aggccttcac cgagacgctg taccccgctg acggcggcct ggaaggcaga 1561 aacgacatgg ccctgaagct cgtgggcggg agccatctga tcgcaaacat caagaccaca 1621 tatagatcca agaaacccgc taagaacctc aagatgcctg gcgtctacta tgtggactac 1681 agactggaaa gaatcaagga ggccaacaac gagacctacg tcgagcagca cgaggtggca 1741 gtggccagat actgcgacct ccctagcaaa ctggggcaca agcttaatta actcgagtct 1801 agagggcccg tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct 1861 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt 1921 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg 1981 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg 2041 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat 2101 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg 2161 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc 2221 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga 2281 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtacc 2341 tagaagttcc tattccgaag ttcctattct ctagaaagta taggaacttc cttggccaaa 2401 aagcctgaac tcaccgcgac gtctgtcgag aagtttctga tcgaaaagtt cgacagcgtc 2461 tccgacctga tgcagctctc ggagggcgaa gaatctcgtg ctttcagctt cgatgtagga 2521 gggcgtggat atgtcctgcg ggtaaatagc tgcgccgatg gtttctacaa agatcgttat 2581 gtttatcggc actttgcatc ggccgcgctc ccgattccgg aagtgcttga cattggggaa 2641 ttcagcgaga gcctgaccta ttgcatctcc cgccgtgcac agggtgtcac gttgcaagac 2701 ctgcctgaaa ccgaactgcc cgctgttctg cagccggtcg cggaggccat ggatgcgatc 2761 gctgcggccg atcttagcca gacgagcggg ttcggcccat tcggaccgca aggaatcggt 2821 caatacacta catggcgtga tttcatatgc gcgattgctg atccccatgt gtatcactgg 2881 caaactgtga tggacgacac cgtcagtgcg tccgtcgcgc aggctctcga tgagctgatg 2941 ctttgggccg aggactgccc cgaagtccgg cacctcgtgc acgcggattt cggctccaac 3001 aatgtcctga cggacaatgg ccgcataaca gcggtcattg actggagcga ggcgatgttc 3061 ggggattccc aatacgaggt cgccaacatc ttcttctgga ggccgtggtt ggcttgtatg 3121 gagcagcaga cgcgctactt cgagcggagg catccggagc ttgcaggatc gccgcggctc 3181 cgggcgtata tgctccgcat tggtcttgac caactctatc agagcttggt tgacggcaat 3241 ttcgatgatg cagcttgggc gcagggtcga tgcgacgcaa tcgtccgatc cggagccggg 3301 actgtcgggc gtacacaaat cgcccgcaga agcgcggccg tctggaccga tggctgtgta 3361 gaagtactcg ccgatagtgg aaaccgacgc cccagcactc gtccgagggc aaaggaatag 3421 cacgtactac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc 3481 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc 3541 gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca 3601 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc 3661 aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg 3721 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc 3781 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg 3841 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc 3901 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact 3961 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta 4021 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag 4081 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc 4141 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta 4201 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg 4261 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc 4321 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac 4381 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac 4441 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg 4501 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga 4561 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt 4621 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag 4681 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct 4741 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg 4801 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat 4861 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc 4921 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg 4981 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct 5041 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca 5101 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg 5161 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg 5221 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc 5281 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag 5341 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg 5401 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag 5461 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat 5521 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg 5581 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca 5641 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca 5701 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat 5761 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag 5821 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtc //