LOCUS V026040 7320 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V026040 VERSION V026040 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 1458..1511 /codon_start=1 /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /product="2A peptide from Thosea asigna virus capsid protein" /protein_id="" /transl_table=1 /translation="EGRGSLLTCGDVEENPGP" /label="T2A" CDS 1530..2000 /label="RBM3(NM_006743)" /note="RBM3(NM_006743)" /gene="RBM3" CDS 2007..2072 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 2096..2684 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 2750..2798 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 3000..3428 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3442..3771 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3838..4632 /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 4808..4929 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4978..4994) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5002..5018 /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(5026..5056) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5071..5092 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5380..5968) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6139..6999) /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(7000..7104) /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 gtggatcgag ggcagaggaa gtcttctaac atgcggtgac gtggaggaga 1501 atcccggccc tgagctcgga tccgccacca tgtcctctga agaaggaaag ctcttcgtgg 1561 gagggctcaa ctttaacacc gacgagcagg cactggaaga ccacttcagc agtttcggac 1621 ctatctctga ggtggtcgtt gtcaaggacc gggagactca gcggtccagg ggttttggtt 1681 tcatcacctt caccaaccca gagcatgctt cagttgccat gagagccatg aacggagagt 1741 ctctggatgg tcgtcagatc cgtgtggatc atgcaggcaa gtctgctcgg ggaaccagag 1801 gaggtggctt tggggcccat gggcgtggtc gcagctactc tagaggtggt ggggaccagg 1861 gctatgggag tggcaggtat tatgacagtc gacctggagg gtatggatat ggatatggac 1921 gttccagaga ctataatggc agaaaccagg gtggttatga ccgctactca ggaggaaatt 1981 acagagacaa ttatgacaac ctcgaggact acaaagacca tgacggtgat tataaagatc 2041 atgacatcga ctacaaggat gacgatgaca agtagtgagg gcccgatatc tcgacaatca 2101 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt 2161 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc 2221 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc 2281 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg 2341 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc 2401 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg 2461 cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg 2521 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc 2581 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct 2641 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggaa acgggggagg 2701 ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa 2761 agacagaata aaacgcacgg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca 2821 gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc 2881 ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt 2941 cggggcggca ggccctgcca tagcagatct gcgcagctgg ggctctaggg ggtatcccca 3001 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc 3061 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac 3121 gttcgccggc tttccccgtc aagctctaaa tcggggcatc cctttagggt tccgatttag 3181 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc 3241 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg 3301 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata 3361 agggattttg gggatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa 3421 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca 3481 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc 3541 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata 3601 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg 3661 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag 3721 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg 3781 ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg 3841 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc 3901 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg 3961 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag 4021 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc 4081 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat 4141 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg 4201 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc 4261 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag 4321 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc 4381 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc 4441 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata 4501 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc 4561 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac 4621 gagttcttct gagcgggact ctggggttcg cgaaatgacc gaccaagcga cgcccaacct 4681 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt 4741 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc 4801 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa 4861 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa 4921 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc 4981 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg 5041 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt 5101 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg 5161 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga 5221 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat 5281 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca 5341 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc 5401 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata 5461 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc 5521 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc 5581 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga 5641 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc 5701 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag 5761 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag 5821 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag 5881 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca 5941 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga 6001 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat 6061 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga 6121 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg 6181 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga 6241 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc 6301 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac 6361 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc 6421 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc 6481 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc 6541 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt 6601 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc 6661 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg 6721 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag 6781 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat 6841 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc 6901 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa 6961 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta 7021 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa 7081 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg 7141 atcgggagat ctcccgatcc cctatggtcg actctcagta caatctgctc tgatgccgca 7201 tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc 7261 aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg //