LOCUS V034381 7804 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V034381 VERSION V034381 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 706..1722 /label="RAD51(NM_002875)" /note="RAD51(NM_002875)" /gene="RAD51" CDS 1753..2460 /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 2467..2556 /codon_start=1 /product="3 tandem Myc epitope tags" /transl_table=1 /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL" /label="3xMyc" misc_feature 2580..3168 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 3234..3282 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 3484..3912 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3926..4255 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4322..5116 /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 5292..5413 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5462..5478) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5486..5502 /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(5510..5540) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5555..5576 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5864..6452) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6623..7483) /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(7484..7588) /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 gctctggcta gcgtttaaac ttaagcttgg 661 taccgagctc ggatccacta gtccagtgtg gtggaattcg ccaccatggc aatgcagatg 721 cagcttgaag caaatgcaga tacttcagtg gaagaagaaa gctttggccc acaacccatt 781 tcacggttag agcagtgtgg cataaatgcc aacgatgtga agaaattgga agaagctgga 841 ttccatactg tggaggctgt tgcctatgcg ccaaagaagg agctaataaa tattaaggga 901 attagtgaag ccaaagctga taaaattctg gctgaggcag ctaaattagt tccaatgggt 961 ttcaccactg caactgaatt ccaccaaagg cggtcagaga tcatacagat tactactggc 1021 tccaaagagc ttgacaaact acttcaaggt ggaattgaga ctggatctat cacagaaatg 1081 tttggagaat tccgaactgg gaagacccag atctgtcata cgctagctgt cacctgccag 1141 cttcccattg accggggtgg aggtgaagga aaggccatgt acattgacac tgagggtacc 1201 tttaggccag aacggctgct ggcagtggct gagaggtatg gtctctctgg cagtgatgtc 1261 ctggataatg tagcatatgc tcgagcgttc aacacagacc accagaccca gctcctttat 1321 caagcatcag ccatgatggt agaatctagg tatgcactgc ttattgtaga cagtgccacc 1381 gccctttaca gaacagacta ctcgggtcga ggtgagcttt cagccaggca gatgcacttg 1441 gccaggtttc tgcggatgct tctgcgactc gctgatgagt ttggtgtagc agtggtaatc 1501 actaatcagg tggtagctca agtggatgga gcagcgatgt ttgctgctga tcccaaaaaa 1561 cctattggag gaaatatcat cgcccatgca tcaacaacca gattgtatct gaggaaagga 1621 agaggggaaa ccagaatctg caaaatctac gactctccct gtcttcctga agctgaagct 1681 atgttcgcca ttaatgcaga tggagtggga gatgccaaag acggtggagg cggatcagga 1741 ggcggtggat ctatggtgag caagggcgag gaggataaca tggccatcat caaggagttc 1801 atgcgcttca aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc 1861 gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt 1921 ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc 1981 tacgtgaagc accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc 2041 aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc 2101 tccctgcagg acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc 2161 gacggccccg taatgcagaa gaagaccatg ggctgggagg cctcctccga gcggatgtac 2221 cccgaggacg gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa ggacggcggc 2281 cactacgacg ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc 2341 gcctacaacg tcaacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg 2401 gaacagtacg aacgcgccga gggccgccac tccaccggcg gcatggacga gctgtacaag 2461 ctcgaggaac aaaaactaat atcagaggaa gatttggaac agaaactaat cagtgaagaa 2521 gatttagagc aaaagttaat ttccgaggag gacttatagt gagggcccga tatctcgaca 2581 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc 2641 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta 2701 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt 2761 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg 2821 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta 2881 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt 2941 tgggcactga caattccgtg gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg 3001 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca 3061 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc 3121 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct ggaaacgggg 3181 gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat 3241 aaaaagacag aataaaacgc acgggtgttg ggtcgtttgt tcataaacgc ggggttcggt 3301 cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg 3361 tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca 3421 acgtcggggc ggcaggccct gccatagcag atctgcgcag ctggggctct agggggtatc 3481 cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga 3541 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg 3601 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg catcccttta gggttccgat 3661 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg 3721 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata 3781 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt 3841 tataagggat tttggggatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat 3901 ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc 3961 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa 4021 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac 4081 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc 4141 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc 4201 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct 4261 cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg 4321 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt 4381 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc 4441 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact 4501 gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt 4561 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca 4621 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat 4681 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg 4741 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga 4801 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga 4861 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa 4921 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga 4981 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt 5041 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct 5101 tgacgagttc ttctgagcgg gactctgggg ttcgcgaaat gaccgaccaa gcgacgccca 5161 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa 5221 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct 5281 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca 5341 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca 5401 tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat 5461 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag 5521 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg 5581 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa 5641 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca 5701 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg 5761 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc 5821 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc 5881 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac 5941 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc 6001 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcaat 6061 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc 6121 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca 6181 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag 6241 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta 6301 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg 6361 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc 6421 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt 6481 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa 6541 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat 6601 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga 6661 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac 6721 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg 6781 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg 6841 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt 6901 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct 6961 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat 7021 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta 7081 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca 7141 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat 7201 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac 7261 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa 7321 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt 7381 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg 7441 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat 7501 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt 7561 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg 7621 acggatcggg agatctcccg atcccctatg gtcgactctc agtacaatct gctctgatgc 7681 cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc 7741 gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct 7801 tagg //