LOCUS Exported 8864 bp ds-DNA circular SYN 31-JAN-2017 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS Untitled 5 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8864) AUTHORS . TITLE Direct Submission JOURNAL Exported Tuesday, Jan 31, 2017 from SnapGene Viewer 3.2.1 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8864 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 4..383 /note="CMV enhancer" /note="human cytomegalovirus immediate early enhancer" promoter 386..661 /note="chicken beta-actin promoter" intron 662..1670 /note="chimeric intron" /note="chimera between introns from chicken beta-actin and rabbit beta-globin" CDS 1742..2689 /codon_start=1 /note="ERT2" /translation="MAGDMRAANLWPSPLMIKRSKKNSLALSLTADQMVSALLDAEPPI LYSEYDPTRPFSEASMMGLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLE ILMIGLVWRSMEHPVKLLFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEE FVCLKSIILLNSGVYTFLSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQR LAQLLLILSHIRHMSNKGMEHLYSMKCKNVVPLYDLLLEAADAHRLHAPTSRGGASVEE TDQSHLATAGSTSSHSLQKYYITGEAEGFPATAVD" misc_feature 2696..3421 /note="PhoCl" misc_feature 3428..4114 /note="Gal4VP16" CDS 3860..3880 /codon_start=1 /product="nuclear localization signal of SV40 large T antigen" /note="SV40 NLS" /translation="PKKKRKV" CDS 3881..4114 /codon_start=1 /gene="UL48" /product="transcriptional activation domain of herpes simplex virus protein VP16 (Triezenberg et al., 1988; Cousens et al., 1989)" /note="VP16 AD" /translation="APPTDVSLGDELHLDGEDVAMAHADALDDFDLDMLGDGDSPGPGF TPHDSAPYGALDMADFEFEQMFTDALGIDEYGG" misc_feature 4121..4846 /note="PhoCl" misc_feature 4853..5797 /note="ERT2" polyA_site 5953..6008 /note="beta-globin poly(A) signal" /note="rabbit beta-globin polyadenylation signal" primer_bind complement(6367..6383) /note="M13 rev" /note="common sequencing primer, one of multiple similar variants" protein_bind 6391..6407 /bound_moiety="lac repressor encoded by lacI" /note="lac operator" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(6415..6445) /note="lac promoter" /note="promoter for the E. coli lac operon" protein_bind 6460..6481 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding site" /note="CAP binding activates transcription in the presence of cAMP." promoter 6540..6735 /note="SV40 promoter" /note="SV40 early promoter" rep_origin 6586..6721 /note="SV40 ori" /note="SV40 origin of replication" polyA_signal 6741..6875 /note="SV40 poly(A) signal" /note="SV40 polyadenylation signal" rep_origin complement(7114..7702) /direction=LEFT /note="ori" /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(7873..8733) /codon_start=1 /gene="bla" /product="beta-lactamase" /note="AmpR" /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" promoter complement(8734..8838) /gene="bla" /note="AmpR promoter" ORIGIN 1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata 61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc 121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag 181 ggactttcca ttgacgtcaa tgggtggact atttacggta aactgcccac ttggcagtac 241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg 301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg 361 tattagtcat cgctattacc atgggtcgag gtgagcccca cgttctgctt cactctcccc 421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca 481 gcgatggggg cggggggggg gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg 541 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt 601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc 661 gggagtcgct gcgttgcctt cgccccgtgc cccgctccgc gccgcctcgc gccgcccgcc 721 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc 781 gggctgtaat tagcgcttgg tttaatgacg gctcgtttct tttctgtggc tgcgtgaaag 841 ccttaaaggg ctccgggagg gccctttgtg cgggggggag cggctcgggg ggtgcgtgcg 901 tgtgtgtgtg cgtggggagc gccgcgtgcg gcccgcgctg cccggcggct gtgagcgctg 961 cgggcgcggc gcggggcttt gtgcgctccg cgtgtgcgcg aggggagcgc ggccgggggc 1021 ggtgccccgc ggtgcggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg 1081 tgggggggtg agcagggggt gtgggcgcgg cggtcgggct gtaacccccc cctgcacccc 1141 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtgc ggggcgtggc 1201 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg 1261 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gccccggagc gccggcggct 1321 gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcagg 1381 gacttccttt gtcccaaatc tggcggagcc gaaatctggg aggcgccgcc gcaccccctc 1441 tagcgggcgc gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt 1501 cgtgcgtcgc cgcgccgccg tccccttctc catctccagc ctcggggctg ccgcaggggg 1561 acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt gtgaccggcg 1621 gctctagagc ctctgctaac catgttcatg ccttcttctt tttcctacag ctcctgggca 1681 acgtgctggt tattgtgctg tctcatcatt ttggcaaaga attcccgggt gagccgccac 1741 catggctgga gacatgagag ctgccaacct ttggccaagc ccgctcatga tcaaacgctc 1801 taagaagaac agcctggcct tgtccctgac ggccgaccag atggtcagtg ccttgttgga 1861 tgctgagccc cccatactct attccgagta tgatcctacc agacccttca gtgaagcttc 1921 gatgatgggc ttactgacca acctggcaga cagggagctg gttcacatga tcaactgggc 1981 gaagagggtg ccaggctttg tggatttgac cctccatgat caggtccacc ttctagaatg 2041 tgcctggcta gagatcctga tgattggtct cgtctggcgc tccatggagc acccagtgaa 2101 gctactgttt gctcctaact tgctcttgga caggaaccag ggaaaatgtg tagagggcat 2161 ggtggagatc ttcgacatgc tgctggctac atcatctcgg ttccgcatga tgaatctgca 2221 gggagaggag tttgtgtgcc tcaaatctat tattttgctt aattctggag tgtacacatt 2281 tctgtccagc accctgaagt ctctggaaga gaaggaccat atccaccgag tcctggacaa 2341 gatcacagac actttgatcc acctgatggc caaggcaggc ctgaccctgc agcagcagca 2401 ccagcggctg gcccagctcc tcctcatcct ctcccacatc aggcacatga gtaacaaagg 2461 catggagcat ctgtacagca tgaagtgcaa gaacgtggtg cccctctatg acctgctgct 2521 ggaggcggcg gacgcccacc gcctacatgc gcccactagc cgtggagggg catccgtgga 2581 ggagacggac caaagccact tggccactgc gggctctact tcatcgcatt ccttgcaaaa 2641 gtattacatc acgggggagg cagagggttt ccctgccaca gctgtcgacg gtaccgtgat 2701 ccctgactac ttcaagcaga gcttccccga gggctacagc tgggagcgca gcatgaccta 2761 cgaggacggc ggcatctgca tcgccaccaa cgacatcaca atggaggggg acagcttcat 2821 caacaagatc cacttcaagg gcacgaactt cccccccaac ggccccgtga tgcagaagag 2881 gaccgtgggc tgggaggcca gcaccgagaa gatgtacgag cgcgacggcg tgctgaaggg 2941 cgacgtgaag atgaagctgc tgctgaaggg cggcggccac tatcgctgcg actaccgcac 3001 cacctacaag gtcaagcaga agcccgtaaa gctgcccgac taccacttcg tggaccaccg 3061 catcgagatc ctgagccacg acaaggacta caacaaggtg aagctgtacg agcacgccgt 3121 ggcccgcaac tccaccgaca gcatggacga gctgtacaag ggtggcagcg gtggcatggt 3181 gagcaagggc gaggagacca ttacaagcgt gatcaagcct gacatgaaga acaagctgcg 3241 catggagggc aacgtgaacg gccacgcctt cgtgatcgag ggcgagggca gcggcaagcc 3301 cttcgagggc atccagacga ttgatttgga ggtgaaggag ggcgccccgc tgcccttcgc 3361 ctacgacatc ctgaccaccg ccttccacta cggcaaccgc gtgttcacca agtacccacg 3421 ggctagctct atcgaacaag catgcgatat ttgccgactt aaaaagctca agtgctccaa 3481 agaaaaaccg aagtgcgcca agtgtctgaa gaacaactgg gagtgtcgct actctcccaa 3541 aaccaaaagg tctccgctga ctagggcaca tctgacagaa gtggaatcaa ggctagaaag 3601 actggaacag ctatttctac tgatttttcc tcgagaagac cttgacatga ttttgaaaat 3661 ggattcttta caggatataa aagcattgtt aacaggatta tttgtacaag ataatgtgaa 3721 taaagatgcc gtcacagata gattggcttc agtggagact gatatgcctc taacattgag 3781 acagcataga ataagtgcga catcatcatc ggaagagagt agtaacaaag gtcaaagaca 3841 gttgactgta tcgatgggcc ctaaaaagaa gcgtaaagtc gcccccccga ccgatgtcag 3901 cctgggggac gagctccact tagacggcga ggacgtggcg atggcgcatg ccgacgcgct 3961 agacgatttc gatctggaca tgttggggga cggggattcc ccgggtccgg gatttacccc 4021 ccacgactcc gccccctacg gcgctctgga tatggccgac ttcgagtttg agcagatgtt 4081 taccgatgcc cttggaattg acgagtacgg tgggtccgga gtgatccctg actacttcaa 4141 gcagagcttc cccgagggct acagctggga gcgcagcatg acctacgagg acggcggcat 4201 ctgcatcgcc accaacgaca tcacaatgga gggggacagc ttcatcaaca agatccactt 4261 caagggcacg aacttccccc ccaacggccc cgtgatgcag aagaggaccg tgggctggga 4321 ggccagcacc gagaagatgt acgagcgcga cggcgtgctg aagggcgacg tgaagatgaa 4381 gctgctgctg aagggcggcg gccactatcg ctgcgactac cgcaccacct acaaggtcaa 4441 gcagaagccc gtaaagctgc ccgactacca cttcgtggac caccgcatcg agatcctgag 4501 ccacgacaag gactacaaca aggtgaagct gtacgagcac gccgtggccc gcaactccac 4561 cgacagcatg gacgagctgt acaagggtgg cagcggtggc atggtgagca agggcgagga 4621 gaccattaca agcgtgatca agcctgacat gaagaacaag ctgcgcatgg agggcaacgt 4681 gaacggccac gccttcgtga tcgagggcga gggcagcggc aagcccttcg agggcatcca 4741 gacgattgat ttggaggtga aggagggcgc cccgctgccc ttcgcctacg acatcctgac 4801 caccgccttc cactacggca accgcgtgtt caccaagtac ccacggctcg acccatctgc 4861 tggagacatg agagctgcca acctttggcc aagcccgctc atgatcaaac gctctaagaa 4921 gaacagcctg gccttgtccc tgacggccga ccagatggtc agtgccttgt tggatgctga 4981 gccccccata ctctattccg agtatgatcc taccagaccc ttcagtgaag cttcgatgat 5041 gggcttactg accaacctgg cagacaggga gctggttcac atgatcaact gggcgaagag 5101 ggtgccaggc tttgtggatt tgaccctcca tgatcaggtc caccttctag aatgtgcctg 5161 gctagagatc ctgatgattg gtctcgtctg gcgctccatg gagcacccag tgaagctact 5221 gtttgctcct aacttgctct tggacaggaa ccagggaaaa tgtgtagagg gcatggtgga 5281 gatcttcgac atgctgctgg ctacatcatc tcggttccgc atgatgaatc tgcagggaga 5341 ggagtttgtg tgcctcaaat ctattatttt gcttaattct ggagtgtaca catttctgtc 5401 cagcaccctg aagtctctgg aagagaagga ccatatccac cgagtcctgg acaagatcac 5461 agacactttg atccacctga tggccaaggc aggcctgacc ctgcagcagc agcaccagcg 5521 gctggcccag ctcctcctca tcctctccca catcaggcac atgagtaaca aaggcatgga 5581 gcatctgtac agcatgaagt gcaagaacgt ggtgcccctc tatgacctgc tgctggaggc 5641 ggcggacgcc caccgcctac atgcgcccac tagccgtgga ggggcatccg tggaggagac 5701 ggaccaaagc cacttggcca ctgcgggctc tacttcatcg cattccttgc aaaagtatta 5761 catcacgggg gaggcagagg gtttccctgc cacagcttga tagcggccgc actcctcagg 5821 tgcaggctgc ctatcagaag gtggtggctg gtgtggccaa tgccctggct cacaaatacc 5881 actgagatct ttttccctct gccaaaaatt atggggacat catgaagccc cttgagcatc 5941 tgacttctgg ctaataaagg aaatttattt tcattgcaat agtgtgttgg aattttttgt 6001 gtctctcact cggaaggaca tatgggaggg caaatcattt aaaacatcag aatgagtatt 6061 tggtttagag tttggcaaca tatgccatat gctggctgcc atgaacaaag gtggctataa 6121 agaggtcatc agtatatgaa acagccccct gctgtccatt ccttattcca tagaaaagcc 6181 ttgacttgag gttagatttt ttttatattt tgttttgtgt tatttttttc tttaacatcc 6241 ctaaaatttt ccttacatgt tttactagcc agatttttcc tcctctcctg actactccca 6301 gtcatagctg tccctcttct cttatgaaga tccctcgacc tgcagcccaa gcttggcgta 6361 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat 6421 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt 6481 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agcggatccg 6541 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact 6601 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag 6661 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc 6721 ctaggctttt gcaaaaagct aacttgttta ttgcagctta taatggttac aaataaagca 6781 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt 6841 ccaaactcat caatgtatct tatcatgtct ggatccgctg cattaatgaa tcggccaacg 6901 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct 6961 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt 7021 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc 7081 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga 7141 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata 7201 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac 7261 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg 7321 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc 7381 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag 7441 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt 7501 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt 7561 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg 7621 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac 7681 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca 7741 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac 7801 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac 7861 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt 7921 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt 7981 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt 8041 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc 8101 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa 8161 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg 8221 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt 8281 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc 8341 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt 8401 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg 8461 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac 8521 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc 8581 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt 8641 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg 8701 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag 8761 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa 8821 acaaataggg gttccgcgca catttccccg aaaagtgcca cctg //