LOCUS V025014 8272 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V025014 VERSION V025014 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 931..3054 /label="MRE11(NM_005591)" /note="MRE11(NM_005591)" /gene="MRE11" CDS 3091..3801 /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" polyA_signal 3839..4063 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 4109..4537 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 4551..4880 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4947..5741 /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 5915..6036 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(6085..6101) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6109..6125 /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(6133..6163) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6178..6199 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6487..7072) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7243..8103) /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(8104..8208) /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 ggctagcgtt taaacttaag cttggtaccg agctcggatc 901 cactagtcca gtgtggtgga attcgccacc atgagtactg cagatgcact tgatgatgaa 961 aacacattta aaatattagt tgcaacagat attcatcttg gatttatgga gaaagatgca 1021 gtcagaggaa atgatacgtt tgtaacactc gatgaaattt taagacttgc ccaggaaaat 1081 gaagtggatt ttattttgtt aggtggtgat ctttttcatg aaaataagcc ctcaaggaaa 1141 acattacata cctgcctcga gttattaaga aaatattgta tgggtgatcg gcctgtccag 1201 tttgaaattc tcagtgatca gtcagtcaac tttggtttta gtaagtttcc atgggtgaac 1261 tatcaagatg gcaacctcaa catttcaatt ccagtgttta gtattcatgg caatcatgac 1321 gatcccacag gggcagatgc actttgtgcc ttggacattt taagttgtgc tggatttgta 1381 aatcactttg gacgttcaat gtctgtggag aagatagaca ttagtccggt tttgcttcaa 1441 aaaggaagca caaagattgc gctatatggt ttaggatcca ttccagatga aaggctctat 1501 cgaatgtttg tcaataaaaa agtaacaatg ttgagaccaa aggaagatga gaactcttgg 1561 tttaacttat ttgtgattca tcagaacagg agtaaacatg gaagtactaa cttcattcca 1621 gaacaatttt tggatgactt cattgatctt gttatctggg gccatgaaca tgagtgtaaa 1681 atagctccaa ccaaaaatga acaacagctg ttttatatct cacaacctgg aagctcagtg 1741 gttacttctc tttccccagg agaagctgta aagaaacatg ttggtttgct gcgtattaaa 1801 gggaggaaga tgaatatgca taaaattcct cttcacacag tgcggcagtt tttcatggag 1861 gatattgttc tagctaatca tccagacatt tttaacccag ataatcctaa agtaacccaa 1921 gccatacaaa gcttctgttt ggagaagatt gaagaaatgc ttgaaaatgc tgaacgggaa 1981 cgtctgggta attctcacca gccagagaag cctcttgtac gactgcgagt ggactatagt 2041 ggaggttttg aacctttcag tgttcttcgc tttagccaga aatttgtgga tcgggtagct 2101 aatccaaaag acattatcca ttttttcagg catagagaac aaaaggaaaa aacaggagaa 2161 gagatcaact ttgggaaact tatcacaaag ccttcagaag gaacaacttt aagggtagaa 2221 gatcttgtaa aacagtactt tcaaaccgca gagaagaatg tgcagctctc actgctaaca 2281 gaaagaggga tgggtgaagc agtacaagaa tttgtggaca aggaggagaa agatgccatt 2341 gaggaattag tgaaatacca gttggaaaaa acacagcgat ttcttaaaga acgtcatatt 2401 gatgccctcg aagacaaaat cgatgaggag gtacgtcgtt tcagagaaac cagacaaaaa 2461 aatactaatg aagaagatga tgaagtccgt gaggctatga ccagggccag agcactcaga 2521 tctcagtcag aggagtctgc ttctgccttt agtgctgatg accttatgag tatagattta 2581 gcagaacaga tggctaatga ctctgatgat agcatctcag cagcaaccaa caaaggaaga 2641 ggccgaggaa gaggtcgaag aggtggaaga gggcagaatt cagcatcgag aggagggtct 2701 caaagaggaa gagcagacac tggtctggag acttctaccc gtagcaggaa ctcaaagact 2761 gctgtgtcag catctagaaa tatgtctatt atagatgcct ttaaatctac aagacagcag 2821 ccttcccgaa atgtcactac taagaattat tcagaggtga ttgaggtaga tgaatcagat 2881 gtggaagaag acatttttcc taccacttca aagacagatc aaaggtggtc cagcacatca 2941 tccagcaaaa tcatgtccca gagtcaagta tcgaaagggg ttgattttga atcaagtgag 3001 gatgatgatg atgatccttt tatgaacact agttctttaa gaagaaatag aagactcgag 3061 ggtggaggcg gatcaggagg cggtggatct atggtgagca agggcgagga ggataacatg 3121 gccatcatca aggagttcat gcgcttcaag gtgcacatgg agggctccgt gaacggccac 3181 gagttcgaga tcgagggcga gggcgagggc cgcccctacg agggcaccca gaccgccaag 3241 ctgaaggtga ccaagggtgg ccccctgccc ttcgcctggg acatcctgtc ccctcagttc 3301 atgtacggct ccaaggccta cgtgaagcac cccgccgaca tccccgacta cttgaagctg 3361 tccttccccg agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg cggcgtggtg 3421 accgtgaccc aggactcctc cctgcaggac ggcgagttca tctacaaggt gaagctgcgc 3481 ggcaccaact tcccctccga cggccccgta atgcagaaga agaccatggg ctgggaggcc 3541 tcctccgagc ggatgtaccc cgaggacggc gccctgaagg gcgagatcaa gcagaggctg 3601 aagctgaagg acggcggcca ctacgacgct gaggtcaaga ccacctacaa ggccaagaag 3661 cccgtgcagc tgcccggcgc ctacaacgtc aacatcaagt tggacatcac ctcccacaac 3721 gaggactaca ccatcgtgga acagtacgaa cgcgccgagg gccgccactc caccggcggc 3781 atggacgagc tgtacaagta gtctagaggg cccgtttaaa cccgctgatc agcctcgact 3841 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg 3901 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg 3961 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg 4021 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga 4081 accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg 4141 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct 4201 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat 4261 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt 4321 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg 4381 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac 4441 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta 4501 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt 4561 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca 4621 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa 4681 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc 4741 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg 4801 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg 4861 gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc 4921 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc 4981 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct 5041 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg 5101 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca 5161 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc 5221 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga 5281 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc 5341 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc 5401 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg 5461 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct 5521 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc 5581 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc 5641 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc 5701 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga 5761 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt 5821 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg 5881 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg 5941 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc 6001 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc 6061 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct 6121 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg 6181 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct 6241 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg 6301 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc 6361 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg 6421 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct 6481 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca 6541 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct 6601 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc 6661 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt 6721 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc 6781 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc 6841 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg 6901 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc 6961 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag 7021 cggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc 7081 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt 7141 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt 7201 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag 7261 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt 7321 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc 7381 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc 7441 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg 7501 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac 7561 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg 7621 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc 7681 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact 7741 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc 7801 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat 7861 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc 7921 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac 7981 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa 8041 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact 8101 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg 8161 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg 8221 aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg gt //