LOCUS V027499 8540 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V027499 VERSION V027499 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" CDS 775..2466 /label="DPYSL5(NM_020134)" /note="DPYSL5(NM_020134)" /gene="DPYSL5" CDS 2482..2538 /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 porcine teschovirus-1 polyprotein" /transl_table=1 /translation="ATNFSLLKQAGDVEENPGP" /label="P2A" CDS 2539..3249 /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" misc_feature 3316..3904 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 3970..4018 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 4220..4648 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 4662..4991 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 5058..5852 /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 6028..6149 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(6198..6214) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6222..6238 /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(6246..6276) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6291..6312 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6600..7188) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7359..8219) /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(8220..8324) /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 gctcgtttag tgaaccgtca gatcgcctgg 661 agacgccatc cacgctgttt tgacctccat agaagacacc gggaccgatc cagcctccgg 721 actctagagc tagcgtttaa acttaagctt ggtaccgagc tcggatccgc caccatgctt 781 gccaactcag ccagcgtgag gatcctcatc aagggaggca aggtggtgaa cgatgactgc 841 acccacgagg ctgacgtcta catcgagaat ggcatcatcc agcaggtggg ccgcgagctc 901 atgatccctg gcggggccaa ggtgattgat gccacaggaa aactggtgat ccctggtggc 961 atcgacacca gcacccactt ccaccagacc ttcatgaatg ccacgtgcgt ggacgacttc 1021 taccatggga ccaaggcagc actcgtcgga ggcaccacca tgatcatcgg ccacgtcctg 1081 cccgacaagg agacctccct tgtggacgct tatgagaagt gccgaggtct ggccgacccc 1141 aaggtctgct gtgattacgc cctccacgtg gggatcacct ggtgggcacc caaggtgaaa 1201 gcagaaatgg agacactggt gagggagaag ggtgtcaact cgttccagat gttcatgacc 1261 tacaaggacc tgtacatgct tcgagacagt gagctgtacc aagtgttgca cgcttgcaag 1321 gacattgggg caatcgcccg cgtccatgct gaaaatgggg agcttgtggc cgagggtgct 1381 aaggaggcac tggatttggg gatcacaggc ccagaaggaa tcgagatcag ccgtccagag 1441 gagctggaag ctgaagccac tcatcgtgtt atcaccattg caaacaggac tcactgtcca 1501 atctacctgg tcaacgtgtc cagtatctcg gctggtgacg ttatcgcagc tgctaagatg 1561 caagggaagg ttgtgctggc ggagaccacc actgcacatg ccacgctgac aggcttacac 1621 tactaccacc aggactggtc ccacgcggct gcctatgtca cggtgcctcc cctgagactg 1681 gacaccaaca cctcaaccta cctcatgagc ctgctggcca atgacactct gaacatcgtg 1741 gcatcagatc accggccttt caccacaaag cagaaagcta tgggcaagga agacttcacc 1801 aagatcccac atggagtgag tggcgtgcag gaccgcatga gcgtcatctg ggagagagga 1861 gtggttggag gaaagatgga tgagaaccgt tttgtggccg ttaccagttc caacgcagct 1921 aagcttctga acctgtatcc ccgcaagggc cgcattattc ccggagccga tgctgatgtg 1981 gtggtgtggg acccagaagc cacaaagacc atctcagcca gcacgcaggt ccagggagga 2041 gacttcaacc tgtatgagaa catgcgctgc cacggcgtgc cactggtcac catcagccgg 2101 gggcgcgtcg tgtatgagaa cggcgtcttc atgtgcgccg agggcaccgg caagttctgt 2161 cccctgaggt ccttcccaga cactgtctac aagaagctgg tccagagaga gaagacttta 2221 aaggttagag gagtggaccg cactccctac ctgggggatg tcgctgttgt cgtgcaccct 2281 gggaaaaaag agatgggaac cccactcgca gacactccta cccggcccgt cacccggcat 2341 gggggcatga gggaccttca cgaatccagc ttcagcctct ctggctctca gatcgatgac 2401 catgttccaa agcgagcttc agctcggatc ctcgctcctc ccggaggcag gtcgagtggc 2461 atttgggaat tcggaagcgg agctactaac ttcagcctgc tgaagcaggc tggcgacgtg 2521 gaggagaacc ctggacctat ggtgagcaag ggcgaggagg ataacatggc catcatcaag 2581 gagttcatgc gcttcaaggt gcacatggag ggctccgtga acggccacga gttcgagatc 2641 gagggcgagg gcgagggccg cccctacgag ggcacccaga ccgccaagct gaaggtgacc 2701 aagggtggcc ccctgccctt cgcctgggac atcctgtccc ctcagttcat gtacggctcc 2761 aaggcctacg tgaagcaccc cgccgacatc cccgactact tgaagctgtc cttccccgag 2821 ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac cgtgacccag 2881 gactcctccc tgcaggacgg cgagttcatc tacaaggtga agctgcgcgg caccaacttc 2941 ccctccgacg gccccgtaat gcagaagaag accatgggct gggaggcctc ctccgagcgg 3001 atgtaccccg aggacggcgc cctgaagggc gagatcaagc agaggctgaa gctgaaggac 3061 ggcggccact acgacgctga ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg 3121 cccggcgcct acaacgtcaa catcaagttg gacatcacct cccacaacga ggactacacc 3181 atcgtggaac agtacgaacg cgccgagggc cgccactcca ccggcggcat ggacgagctg 3241 tacaagtaac tcgagtctag agggcccctt aagggttcga tccctaccgg ttagtaatga 3301 gtttgatatc tcgacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt 3361 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat 3421 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct 3481 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct 3541 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc 3601 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg 3661 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc 3721 tttccatggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac 3781 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg 3841 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc 3901 ccgcctggaa acgggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc 3961 gcgctatgac ggcaataaaa agacagaata aaacgcacgg gtgttgggtc gtttgttcat 4021 aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg 4081 ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc 4141 ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcagatct gcgcagctgg 4201 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg 4261 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc 4321 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcggggcatc 4381 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt 4441 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag 4501 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg 4561 gtctattctt ttgatttata agggattttg gggatttcgg cctattggtt aaaaaatgag 4621 ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg 4681 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag 4741 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc 4801 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc 4861 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg 4921 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag 4981 gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag 5041 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt 5101 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg 5161 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg 5221 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg 5281 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg 5341 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca 5401 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc 5461 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc 5521 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca 5581 aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga 5641 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg 5701 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg 5761 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg 5821 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg cgaaatgacc 5881 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa 5941 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat 6001 ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa 6061 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt 6121 ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag 6181 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt 6241 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc 6301 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc 6361 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct 6421 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca 6481 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac 6541 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt 6601 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg 6661 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc 6721 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc 6781 gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc 6841 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac 6901 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt 6961 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct 7021 aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc 7081 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt 7141 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg 7201 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc 7261 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa 7321 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag 7381 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg 7441 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga 7501 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag 7561 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa 7621 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc 7681 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca 7741 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg 7801 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat 7861 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc 7921 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg 7981 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg 8041 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt 8101 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca 8161 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata 8221 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac 8281 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa 8341 gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtcg actctcagta 8401 caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg 8461 tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt 8521 gcatgaagaa tctgcttagg //