LOCUS Exported 5895 bp ds-DNA circular SYN 24-MAY-2017 DEFINITION synthetic circular DNA. ACCESSION . VERSION . KEYWORDS pKK-TEV-mCerulean SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5895) 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..5895 /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..1809 /codon_start=1 /product="Cerulean" /note="mCerulean" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAISDNVYITADKQKNGIK ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" polyA_signal 1853..2077 /note="BGH poly(A)signal" /note="bovine growth hormone polyadenylation signal" protein_bind 2361..2408 /bound_moiety="FLP recombinase" /note="FRT" CDS 2416..3438 /codon_start=1 /product="aminoglycoside phosphotransferase from E. coli" /note="Hygromycin B" /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE " polyA_signal 3568..3698 /note="SV40 poly(A)signal" /note="SV40 polyadenylation signal" rep_origin complement(4081..4754) /direction=LEFT /note="pUC ori" CDS complement(4899..5759) /codon_start=1 /product="beta-lactamase" /note="AmpR" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" misc_feature complement(5760..5858) /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 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc 1141 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat 1201 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc 1261 tggcccaccc tcgtgaccac cctgacctgg ggcgtgcagt gcttcgcccg ctaccccgac 1321 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc 1381 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc 1441 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc 1501 ctggggcaca agctggagta caacgccatc agcgacaacg tctatatcac cgccgacaag 1561 cagaagaacg gcatcaaggc caacttcaag atccgccaca acatcgagga cggcagcgtg 1621 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc 1681 gacaaccact acctgagcac ccagtccaag ctgagcaaag accccaacga gaagcgcgat 1741 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg 1801 tacaagtaac tcgagtctag agggcccgtt taaacccgct gatcagcctc gactgtgcct 1861 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt 1921 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg 1981 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac 2041 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc 2101 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg 2161 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct 2221 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg 2281 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag 2341 ggtgatggtt cacgtaccta gaagttccta ttccgaagtt cctattctct agaaagtata 2401 ggaacttcct tggccaaaaa gcctgaactc accgcgacgt ctgtcgagaa gtttctgatc 2461 gaaaagttcg acagcgtctc cgacctgatg cagctctcgg agggcgaaga atctcgtgct 2521 ttcagcttcg atgtaggagg gcgtggatat gtcctgcggg taaatagctg cgccgatggt 2581 ttctacaaag atcgttatgt ttatcggcac tttgcatcgg ccgcgctccc gattccggaa 2641 gtgcttgaca ttggggaatt cagcgagagc ctgacctatt gcatctcccg ccgtgcacag 2701 ggtgtcacgt tgcaagacct gcctgaaacc gaactgcccg ctgttctgca gccggtcgcg 2761 gaggccatgg atgcgatcgc tgcggccgat cttagccaga cgagcgggtt cggcccattc 2821 ggaccgcaag gaatcggtca atacactaca tggcgtgatt tcatatgcgc gattgctgat 2881 ccccatgtgt atcactggca aactgtgatg gacgacaccg tcagtgcgtc cgtcgcgcag 2941 gctctcgatg agctgatgct ttgggccgag gactgccccg aagtccggca cctcgtgcac 3001 gcggatttcg gctccaacaa tgtcctgacg gacaatggcc gcataacagc ggtcattgac 3061 tggagcgagg cgatgttcgg ggattcccaa tacgaggtcg ccaacatctt cttctggagg 3121 ccgtggttgg cttgtatgga gcagcagacg cgctacttcg agcggaggca tccggagctt 3181 gcaggatcgc cgcggctccg ggcgtatatg ctccgcattg gtcttgacca actctatcag 3241 agcttggttg acggcaattt cgatgatgca gcttgggcgc agggtcgatg cgacgcaatc 3301 gtccgatccg gagccgggac tgtcgggcgt acacaaatcg cccgcagaag cgcggccgtc 3361 tggaccgatg gctgtgtaga agtactcgcc gatagtggaa accgacgccc cagcactcgt 3421 ccgagggcaa aggaatagca cgtactacga gatttcgatt ccaccgccgc cttctatgaa 3481 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat 3541 ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa 3601 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt 3661 ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag 3721 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt 3781 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc 3841 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc 3901 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct 3961 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca 4021 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac 4081 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt 4141 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg 4201 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc 4261 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc 4321 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc 4381 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac 4441 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt 4501 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct 4561 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc 4621 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt 4681 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg 4741 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc 4801 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa 4861 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag 4921 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg 4981 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga 5041 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag 5101 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa 5161 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc 5221 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca 5281 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg 5341 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat 5401 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc 5461 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg 5521 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg 5581 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt 5641 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca 5701 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata 5761 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac 5821 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa 5881 gtgccacctg acgtc //