LOCUS V028276 9188 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V028276 VERSION V028276 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers enhancer 202..581 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 582..785 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" promoter 830..848 /note="promoter for bacteriophage T7 RNA polymerase" /label="T7 promoter" CDS 872..1579 /codon_start=1 /note="mammalian codon-optimized" /product="monomeric derivative of DsRed fluorescent protein (Shaner et al., 2004)" /transl_table=1 /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA EGRHSTGGMDELYK" /label="mCherry" CDS 1616..4819 /label="PIK3CA(NM_006218)" /note="PIK3CA(NM_006218)" /gene="PIK3CA" CDS 4826..4852 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" CDS 4856..4882 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" CDS 4886..4912 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" polyA_signal 4950..5174 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 5220..5648 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 5662..5991 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 6058..6657 /codon_start=1 /gene="pac from Streptomyces alboniger" /note="confers resistance to puromycin" /product="puromycin N-acetyltransferase" /transl_table=1 /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*" /label="PuroR" polyA_signal 6831..6952 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(7001..7017) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7025..7041 /bound_moiety="lac repressor encoded by lacI" /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-β-D-thiogalactopyranoside (IPTG)." /label="lac operator" promoter complement(7049..7079) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7094..7115 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7403..7988) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8159..9019) /codon_start=1 /gene="bla" /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /product="β-lactamase" /transl_table=1 /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA SLIKHW*" /label="AmpR" promoter complement(9020..9124) /gene="bla" /label="AmpR promoter" ORIGIN 1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt 61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct 121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg 181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta 241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg 301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc 361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa 421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca 481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta 541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt 601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg 661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca 721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca 781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc 841 actataggga gacccaagct ggctagccac catggtgagc aagggcgagg aggataacat 901 ggccatcatc aaggagttca tgcgcttcaa ggtgcacatg gagggctccg tgaacggcca 961 cgagttcgag atcgagggcg agggcgaggg ccgcccctac gagggcaccc agaccgccaa 1021 gctgaaggtg accaagggtg gccccctgcc cttcgcctgg gacatcctgt cccctcagtt 1081 catgtacggc tccaaggcct acgtgaagca ccccgccgac atccccgact acttgaagct 1141 gtccttcccc gagggcttca agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt 1201 gaccgtgacc caggactcct ccctgcagga cggcgagttc atctacaagg tgaagctgcg 1261 cggcaccaac ttcccctccg acggccccgt aatgcagaag aagaccatgg gctgggaggc 1321 ctcctccgag cggatgtacc ccgaggacgg cgccctgaag ggcgagatca agcagaggct 1381 gaagctgaag gacggcggcc actacgacgc tgaggtcaag accacctaca aggccaagaa 1441 gcccgtgcag ctgcccggcg cctacaacgt caacatcaag ttggacatca cctcccacaa 1501 cgaggactac accatcgtgg aacagtacga acgcgccgag ggccgccact ccaccggcgg 1561 catggacgag ctgtacaagg gaggtggagg tagtggagga ggcggatcaa gcttcatgcc 1621 tccacgacca tcatcaggtg aactgtgggg catccacttg atgcccccaa gaatcctagt 1681 agaatgttta ctaccaaatg gaatgatagt gactttagaa tgcctccgtg aggctacatt 1741 aataaccata aagcatgaac tatttaaaga agcaagaaaa taccccctcc atcaacttct 1801 tcaagatgaa tcttcttaca ttttcgtaag tgttactcaa gaagcagaaa gggaagaatt 1861 ttttgatgaa acaagacgac tttgtgacct tcggcttttt caaccctttt taaaagtaat 1921 tgaaccagta ggcaaccgtg aagaaaagat cctcaatcga gaaattggtt ttgctatcgg 1981 catgccagtg tgtgaatttg atatggttaa agatccagaa gtacaggact tccgaagaaa 2041 tattctgaac gtttgtaaag aagctgtgga tcttagggac ctcaattcac ctcatagtag 2101 agcaatgtat gtctatcctc caaatgtaga atcttcacca gaattgccaa agcacatata 2161 taataaatta gataaagggc aaataatagt ggtgatctgg gtaatagttt ctccaaataa 2221 tgacaagcag aagtatactc tgaaaatcaa ccatgactgt gtaccagaac aagtaattgc 2281 tgaagcaatc aggaaaaaaa ctcgaagtat gttgctatcc tctgaacaac taaaactctg 2341 tgttttagaa tatcagggca agtatatttt aaaagtgtgt ggatgtgatg aatacttcct 2401 agaaaaatat cctctgagtc agtataagta tataagaagc tgtataatgc ttgggaggat 2461 gcccaatttg atgttgatgg ctaaagaaag cctttattct caactgccaa tggactgttt 2521 tacaatgcca tcttattcca gacgcatttc cacagctaca ccatatatga atggagaaac 2581 atctacaaaa tccctttggg ttataaatag tgcactcaga ataaaaattc tttgtgcaac 2641 ctacgtgaat gtaaatattc gagacattga taagatctat gttcgaacag gtatctacca 2701 tggaggagaa cccttatgtg acaatgtgaa cactcaaaga gtaccttgtt ccaatcccag 2761 gtggaatgaa tggctgaatt atgatatata cattcctgat cttcctcgtg ctgctcgact 2821 ttgcctttcc atttgctctg ttaaaggccg aaagggtgct aaagaggaac actgtccatt 2881 ggcatgggga aatataaact tgtttgatta cacagacact ctagtatctg gaaaaatggc 2941 tttgaatctt tggccagtac ctcatggatt agaagatttg ctgaacccta ttggtgttac 3001 tggatcaaat ccaaataaag aaactccatg cttagagttg gagtttgact ggttcagcag 3061 tgtggtaaag ttcccagata tgtcagtgat tgaagagcat gccaattggt ctgtatcccg 3121 agaagcagga tttagctatt cccacgcagg actgagtaac agactagcta gagacaatga 3181 attaagggaa aatgacaaag aacagctcaa agcaatttct acacgagatc ctctctctga 3241 aatcactgag caggagaaag attttctatg gagtcacaga cactattgtg taactatccc 3301 cgaaattcta cccaaattgc ttctgtctgt taaatggaat tctagagatg aagtagccca 3361 gatgtattgc ttggtaaaag attggcctcc aatcaaacct gaacaggcta tggaacttct 3421 ggactgtaat tacccagatc ctatggttcg aggttttgct gttcggtgct tggaaaaata 3481 tttaacagat gacaaacttt ctcagtattt aattcagcta gtacaggtcc taaaatatga 3541 acaatatttg gataacttgc ttgtgagatt tttactgaag aaagcattga ctaatcaaag 3601 gattgggcac tttttctttt ggcatttaaa atctgagatg cacaataaaa cagttagcca 3661 gaggtttggc ctgcttttgg agtcctattg tcgtgcatgt gggatgtatt tgaagcacct 3721 gaataggcaa gtcgaggcaa tggaaaagct cattaactta actgacattc tcaaacagga 3781 gaagaaggat gaaacacaaa aggtacagat gaagttttta gttgagcaaa tgaggcgacc 3841 agatttcatg gatgctctac agggctttct gtctcctcta aaccctgctc atcaactagg 3901 aaacctcagg cttgaagagt gtcgaattat gtcctctgca aaaaggccac tgtggttgaa 3961 ttgggagaac ccagacatca tgtcagagtt actgtttcag aacaatgaga tcatctttaa 4021 aaatggggat gatttacggc aagatatgct aacacttcaa attattcgta ttatggaaaa 4081 tatctggcaa aatcaaggtc ttgatcttcg aatgttacct tatggttgtc tgtcaatcgg 4141 tgactgtgtg ggacttattg aggtggtgcg aaattctcac actattatgc aaattcagtg 4201 caaaggcggc ttgaaaggtg cactgcagtt caacagccac acactacatc agtggctcaa 4261 agacaagaac aaaggagaaa tatatgatgc agccattgac ctgtttacac gttcatgtgc 4321 tggatactgt gtagctacct tcattttggg aattggagat cgtcacaata gtaacatcat 4381 ggtgaaagac gatggacaac tgtttcatat agattttgga cactttttgg atcacaagaa 4441 gaaaaaattt ggttataaac gagaacgtgt gccatttgtt ttgacacagg atttcttaat 4501 agtgattagt aaaggagccc aagaatgcac aaagacaaga gaatttgaga ggtttcagga 4561 gatgtgttac aaggcttatc tagctattcg acagcatgcc aatctcttca taaatctttt 4621 ctcaatgatg cttggctctg gaatgccaga actacaatct tttgatgaca ttgcatacat 4681 tcgaaagacc ctagccttag ataaaactga gcaagaggct ttggagtatt tcatgaaaca 4741 aatgaatgat gcacatcatg gtggctggac aacaaaaatg gattggatct tccacacaat 4801 taaacagcat gcattgaacc tcgagtaccc atacgacgta cctgattatg caacttatcc 4861 ttatgatgta cctgattatg ctagctatcc ttatgatgtc ccagactacg catagtgagg 4921 gcccgtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt 4981 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta 5041 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg 5101 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc 5161 ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca 5221 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc 5281 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac 5341 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag 5401 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc 5461 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg 5521 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata 5581 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa 5641 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca 5701 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc 5761 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata 5821 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg 5881 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag 5941 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg 6001 ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg 6061 accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtcccccg ggccgtacgc 6121 accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga cccggaccgc 6181 cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc 6241 ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc 6301 gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc 6361 cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc caaggagccc 6421 gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc agggcaaggg tctgggcagc 6481 gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag 6541 acctccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac 6601 gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg tgcctgagcg 6661 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg 6721 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct 6781 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta 6841 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat 6901 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct 6961 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt 7021 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag 7081 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt 7141 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag 7201 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg 7261 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat 7321 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta 7381 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa 7441 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc 7501 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt 7561 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca 7621 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg 7681 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat 7741 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta 7801 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct 7861 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac 7921 aaaccaccgc tggtagcggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag 7981 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact 8041 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa 8101 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt 8161 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag 8221 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca 8281 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc 8341 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt 8401 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg 8461 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca 8521 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg 8581 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca 8641 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg 8701 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct 8761 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca 8821 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca 8881 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg 8941 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac 9001 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt 9061 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc 9121 cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc 9181 cctatggt //