LOCUS V025988 9420 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V025988 VERSION V025988 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers enhancer 50..429 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 430..633 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" promoter 678..696 /note="promoter for bacteriophage T7 RNA polymerase" /label="T7 promoter" CDS 744..1451 /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 1500..4100 /label="E2F8(NM_001256372)" /note="E2F8(NM_001256372)" /gene="E2F8" CDS 4107..4172 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 4196..4784 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 4850..4898 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 5100..5528 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 5542..5871 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 5938..6732 /codon_start=1 /gene="aph(3')-II (or nptII)" /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin®)" /product="aminoglycoside phosphotransferase from Tn5" /transl_table=1 /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*" /label="NeoR/KanR" polyA_signal 6908..7029 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(7078..7094) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7102..7118 /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(7126..7156) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7171..7192 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7480..8068) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8239..9099) /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(9100..9204) /gene="bla" /label="AmpR promoter" ORIGIN 1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta 61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag 121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc 181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga 241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat 301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc 361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct 421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca 481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat 541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg 601 cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga acccactgct 661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta 721 aacttaagct tggtaccgcc accatggtga gcaagggcga ggaggataac atggccatca 781 tcaaggagtt catgcgcttc aaggtgcaca tggagggctc cgtgaacggc cacgagttcg 841 agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg 901 tgaccaaggg tggccccctg cccttcgcct gggacatcct gtcccctcag ttcatgtacg 961 gctccaaggc ctacgtgaag caccccgccg acatccccga ctacttgaag ctgtccttcc 1021 ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga 1081 cccaggactc ctccctgcag gacggcgagt tcatctacaa ggtgaagctg cgcggcacca 1141 acttcccctc cgacggcccc gtaatgcaga agaagaccat gggctgggag gcctcctccg 1201 agcggatgta ccccgaggac ggcgccctga agggcgagat caagcagagg ctgaagctga 1261 aggacggcgg ccactacgac gctgaggtca agaccaccta caaggccaag aagcccgtgc 1321 agctgcccgg cgcctacaac gtcaacatca agttggacat cacctcccac aacgaggact 1381 acaccatcgt ggaacagtac gaacgcgccg agggccgcca ctccaccggc ggcatggacg 1441 agctgtacaa gtggatcgga ggagggggaa gcggtggagg cggtagtgag ctcggatcca 1501 tggagaacga aaaggaaaat ctcttttgtg agccacataa aaggggacta atgaaaacac 1561 ctctgaaaga atccaccaca gcaaatatcg tgttggcaga gatccagcct gactttggcc 1621 ctttaaccac acctaccaag cccaaggaag gctctcaggg agagccgtgg acaccgacag 1681 ccaacctgaa aatgctcatc agtgctgtga gccctgagat ccgcaacaga gatcagaaaa 1741 ggggtttgtt tgacaacaga agtggattac ctgaggccaa agactgtata cacgaacact 1801 tatctggaga tgaatttgag aaatcccaac caagtcgaaa agagaaaagt ttaggattat 1861 tgtgtcataa gttcttagca cgatatccta attatcccaa ccctgctgtg aataatgaca 1921 tctgccttga cgaagtggca gaggaactta atgttgaacg tcgacgcatt tacgatatcg 1981 tgaacgtcct agagagttta catatggtga gccgcctcgc caaaaacagg tacacttggc 2041 acgggcgaca caatctcaac aaaacccttg gcaccttgaa gagcatcggg gaggagaata 2101 agtacgccga gcagattatg atgatcaaaa agaaagaata tgagcaagag tttgacttta 2161 ttaagagtta cagtatagag gatcatatca tcaaatcaaa cactggccca aatggacacc 2221 cagacatgtg ttttgtggaa ctccctggag tggaatttcg ggcagcttct gtaaacagcc 2281 gcaaagacaa gtctttaagg gtaatgagcc agaaatttgt gatgctgttt ttggtgtcaa 2341 cgcctcagat agtaagccta gaagttgctg ccaagatttt aattggggag gaccatgtgg 2401 aagatttgga taaaagcaag tttaaaacaa aaattaggag gttgtatgat atagctaatg 2461 ttctgagtag cctggatctt atcaagaaag ttcatgttac agaggaaaga ggccgaaaac 2521 cagctttcaa atggaccggc ccagaaatca gtccaaatac cagtggctcc agcccagtca 2581 ttcattttac tccctctgat ttggaggtga gacggtcttc aaaagagaac tgtgccaaaa 2641 acctcttttc cacacgtggg aaaccaaact ttactcgaca cccatctctt atcaaattgg 2701 taaagagtat agaaagtgat cggagaaaga taaattctgc gcccagtagc cctatcaaga 2761 ccaacaaagc tgagagttct cagaattctg cacccttccc aagtaaaatg gctcagctcg 2821 cagctatttg taaaatgcag ttagaagagc aatcaagtga atccagacag aaagtgaaag 2881 tacagctggc aagatctgga ccctgcaaac cagtagcccc tctggacccc ccagtgaatg 2941 ctgagatgga gctgacagca ccgtccctca tccagcccct gggaatggtt cccctgatcc 3001 ccagcccctt gtcatcagca gtgcccctga tcctacctca ggccccttca ggcccatcct 3061 atgccatcta cctgcagccc actcaagccc accaaagtgt gacgccaccc caaggcctga 3121 gcccaacggt gtgcaccacc cactcttcta aagctactgg ctcaaaagac tccacagatg 3181 ccaccactga gaaggcagcc aatgatacct caaaggccag tgcctctacc aggcctggaa 3241 gcttgctgcc agcaccagag aggcaagggg caaagagccg aaccagggag ccagctggag 3301 aaagaggctc aaagagggca agcatgctcg aggacagtgg ttccaaaaag aaatttaaag 3361 aggacctaaa aggacttgaa aatgtctccg caaccttgtt cccatcagga tacctaatcc 3421 ctctcacgca gtgctcatcc ctgggggcag agtccatttt gtctggtaaa gaaaactcaa 3481 gtgctctttc cccaaaccac aggatttaca gctccccaat tgcaggtgtt attccagtga 3541 catcatctga actcactgct gttaattttc cctcttttca tgtaacaccg ttgaagctaa 3601 tggtctcacc aacttccgtg gcagccgtac ctgtcgggaa cagcccggct ctcgcttcaa 3661 gccaccctgt tcccatccag aacccaagct cagccattgt aaacttcacc ctgcagcacc 3721 tgggactcat ctcacccaat gtgcagttgt ctgccagccc tgggtctgga atcgttcctg 3781 tgtctccaag aatagagtct gttaatgtcg caccagaaaa tgcaggcact cagcaaggaa 3841 gggccaccaa ctatgactca ccagtcccag gccagagcca gccaaatgga caatcagttg 3901 ctgtgacagg ggcacaacag cctgttcctg tgacacccaa agggtcacaa ttagtggccg 3961 aaagtttctt ccgtacccca ggtggaccca ccaagccaac cagctcatcc tgcatggatt 4021 ttgagggtgc taataaaacc tccttaggaa ctctctttgt cccacagcga aaactggaag 4081 tctcaacaga ggatgtccat ctcgaggact acaaagacca tgacggtgat tataaagatc 4141 atgacatcga ctacaaggat gacgatgaca agtagtgagg gcccgatatc tcgacaatca 4201 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt 4261 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc 4321 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc 4381 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg 4441 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc 4501 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg 4561 cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg 4621 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc 4681 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct 4741 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggaa acgggggagg 4801 ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa 4861 agacagaata aaacgcacgg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca 4921 gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc 4981 ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt 5041 cggggcggca ggccctgcca tagcagatct gcgcagctgg ggctctaggg ggtatcccca 5101 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc 5161 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac 5221 gttcgccggc tttccccgtc aagctctaaa tcggggcatc cctttagggt tccgatttag 5281 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc 5341 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg 5401 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata 5461 agggattttg gggatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa 5521 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca 5581 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc 5641 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata 5701 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg 5761 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag 5821 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg 5881 ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg 5941 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc 6001 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg 6061 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag 6121 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc 6181 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat 6241 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg 6301 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc 6361 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag 6421 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc 6481 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc 6541 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata 6601 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc 6661 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac 6721 gagttcttct gagcgggact ctggggttcg cgaaatgacc gaccaagcga cgcccaacct 6781 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt 6841 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc 6901 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa 6961 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa 7021 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc 7081 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg 7141 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt 7201 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg 7261 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga 7321 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat 7381 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca 7441 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc 7501 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata 7561 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc 7621 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc 7681 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga 7741 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc 7801 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag 7861 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag 7921 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag 7981 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca 8041 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga 8101 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat 8161 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga 8221 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg 8281 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga 8341 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc 8401 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac 8461 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc 8521 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc 8581 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc 8641 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt 8701 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc 8761 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg 8821 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag 8881 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat 8941 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc 9001 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa 9061 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta 9121 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa 9181 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg 9241 atcgggagat ctcccgatcc cctatggtcg actctcagta caatctgctc tgatgccgca 9301 tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc 9361 aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg //