LOCUS V025859 9402 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V025859 VERSION V025859 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..4082 /label="ORC1(NM_004153)" /note="ORC1(NM_004153)" /gene="ORC1" CDS 4089..4154 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 4178..4766 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 4832..4880 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 5082..5510 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 5524..5853 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 5920..6714 /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 6890..7011 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(7060..7076) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7084..7100 /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(7108..7138) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7153..7174 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7462..8050) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8221..9081) /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(9082..9186) /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 tggcacacta ccccacaagg ctgaagacca gaaaaactta ttcatgggtt ggcaggccct 1561 tgttggatcg aaaactgcac taccaaacct atagagaaat gtgtgtgaaa acagaaggtt 1621 gttccaccga gattcacatc cagattggac agtttgtgtt gattgaaggg gatgatgatg 1681 aaaacccgta tgttgctaaa ttgcttgagt tgttcgaaga tgactctgat cctcctccta 1741 agaaacgtgc tcgagtacag tggtttgtcc gattctgtga agtccctgcc tgtaaacggc 1801 atttgttggg ccggaagcct ggtgcacagg aaatattctg gtatgattac ccggcctgtg 1861 acagcaacat taatgcggag accatcattg gccttgttcg ggtgatacct ttagccccaa 1921 aggatgtggt accgacgaat ctgaaaaatg agaagacact ctttgtgaaa ctatcctgga 1981 atgagaagaa attcaggcca ctttcctcag aactatttgc ggagttgaat aaaccacaag 2041 agagtgcagc caagtgccag aaacccgtga gagccaagag taagagtgca gagagccctt 2101 cttggacccc agcagaacat gtggccaaaa ggattgaatc aaggcactcc gcctccaaat 2161 ctcgccaaac tcctacccat cctcttaccc caagagccag aaagaggctg gagcttggca 2221 acttaggtaa ccctcagatg tcccagcaga cttcatgtgc ctccttggat tctccaggaa 2281 gaataaaacg gaaagtggcc ttctcggaga tcacctcacc ttctaagaga tctcagcctg 2341 ataaacttca aaccttgtct ccagctctga aagccccaga gaaaaccaga gagactggac 2401 tctcttatac tgaggatgac aagaaggctt cacctgaaca tcgcataatc ctgagaaccc 2461 gaattgcagc ttcgaaaacc atagacatta gagaggagag aacacttacc cctatcagtg 2521 ggggacagag atcttcagtg gtgccatccg tgattctgaa accagaaaac atcaaaaaga 2581 gggatgcaaa agaagcaaaa gcccagaatg aagcgacctc tactccccat cgtatccgca 2641 gaaagagttc tgtcttgact atgaatcgga ttaggcagca gcttcggttt ctaggtaata 2701 gtaaaagtga ccaagaagag aaagagattc tgccagcagc agagatttca gactctagca 2761 gtgacgaaga agaggcttcc acaccgcccc ttccaaggag agcacccaga actgtgtcca 2821 ggaacctgcg atcttccttg aagtcatcct tacataccct cacgaaggtg ccaaagaaga 2881 gtctcaagcc tagaacgcca cgttgtgccg ctcctcagat ccgtagtcga agcctggctg 2941 cccaggagcc agccagtgtg ctggaggaag cccgactgag gctgcatgtt tctgctgtac 3001 ctgagtctct tccctgtcgg gaacaggaat tccaagacat ctacaatttt gtggaaagca 3061 aactccttga ccataccgga gggtgcatgt acatctccgg tgtccctggg acagggaaga 3121 ctgccactgt tcatgaagtg atacgctgcc tgcagcaggc agcccaagcc aatgatgttc 3181 ctccctttca atacattgag gtcaatggca tgaagctgac ggagccccac caagtctatg 3241 tgcaaatctt gcagaagcta acaggccaaa aagcaacagc caaccatgcg gcagaactgc 3301 tggcaaagca attctgcacc cgagggtcac ctcaggaaac caccgtcctg cttgtggatg 3361 agctcgacct tctgtggact cacaaacaag acataatgta caatctcttt gactggccca 3421 ctcataagga ggcccggctt gtggtcctgg caattgccaa cacaatggac ctgccagagc 3481 gaatcatgat gaaccgggtg tccagccgac tgggtcttac caggatgtgc ttccagccct 3541 atacatatag ccagctgcag cagatcctaa ggtcccggct caagcatcta aaggcctttg 3601 aagatgatgc catccagctg gtagccagga aggtagcagc actgtctgga gatgcacgac 3661 ggtgcctgga catctgcagg cgtgccacag agatctgtga gttctcccag cagaagcctg 3721 actcccctgg cctggtcacc atagcccact caatggaagc tgtggatgag atgttttcat 3781 catcatacat cacggccatc aaaaattcct ctgttctgga acagagcttc ctgagagcca 3841 tcctcgcaga gttccgtcga tcaggactgg aggaagccac gtttcaacag atatatagtc 3901 aacatgtggc actgtgcaga atggagggac tgccgtaccc caccatgtca gagaccatgg 3961 ccgtgtgttc tcacctgggc tcctgtcgcc tcctgcttgt ggagcccagc aggaacgatc 4021 tgctccttcg ggtgcggctc aacgtcagcc aggatgatgt gctgtatgcg ctgaaagacg 4081 agctcgagga ctacaaagac catgacggtg attataaaga tcatgacatc gactacaagg 4141 atgacgatga caagtagtga gggcccgata tctcgacaat caacctctgg attacaaaat 4201 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc 4261 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt 4321 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg 4381 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg 4441 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc 4501 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt 4561 gttgtcgggg aagctgacgt cctttccatg gctgctcgcc tgtgttgcca cctggattct 4621 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg 4681 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg 4741 gatctccctt tgggccgcct ccccgcctgg aaacggggga ggctaactga aacacggaag 4801 gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac 4861 gggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga 4921 taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca 4981 ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc 5041 catagcagat ctgcgcagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc 5101 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct 5161 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg 5221 tcaagctcta aatcggggca tccctttagg gttccgattt agtgctttac ggcacctcga 5281 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt 5341 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg 5401 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tggggatttc 5461 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg 5521 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa 5581 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc 5641 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg 5701 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt 5761 ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga 5821 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt 5881 ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg 5941 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag 6001 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt 6061 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta 6121 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg 6181 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt 6241 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat 6301 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg 6361 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca 6421 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc 6481 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc 6541 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat 6601 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc 6661 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga 6721 ctctggggtt cgcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga 6781 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg 6841 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat 6901 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt 6961 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg 7021 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg 7081 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc 7141 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt 7201 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg 7261 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt 7321 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc 7381 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa 7441 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa 7501 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc 7561 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc 7621 cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag 7681 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga 7741 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc 7801 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac 7861 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg 7921 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca 7981 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa 8041 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa 8101 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt 8161 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag 8221 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat 8281 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc 8341 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa 8401 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca 8461 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa 8521 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt 8581 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc 8641 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact 8701 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc 8761 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg 8821 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct 8881 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc 8941 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag 9001 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac 9061 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg 9121 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt 9181 tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat 9241 cccctatggt cgactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg 9301 ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca 9361 aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gg //