LOCUS V028781 6829 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V028781 VERSION V028781 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers enhancer 235..614 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 615..818 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" promoter 863..881 /note="promoter for bacteriophage T7 RNA polymerase" /label="T7 promoter" CDS 964..2319 /label="Idh2(NM_173011)" /note="Idh2(NM_173011)" /gene="Idh2" CDS 2326..2391 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 2429..2653 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2699..3127 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3141..3470 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3537..4331 /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 4505..4626 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4675..4691) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4699..4715 /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(4723..4753) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4768..4789 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5077..5662) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5833..6693) /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(6694..6798) /gene="bla" /label="AmpR promoter" ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg 61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt 241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata 301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc 361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc 421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt 481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt 541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca 601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg 661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc 721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg 781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca 841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc 901 gtttaaactt aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattcgcc 961 accatggccg gctacctgcg ggctgtgagc tcgctctgca gagcctcggg ctcagcgcgg 1021 acctgggcac cggcagcact gactgtcccc agctggccgg agcagccgcg gcgccactat 1081 gctgagaaga ggatcaaggt ggagaagccg gtagtggaga tggacggtga cgagatgacc 1141 cggatcatct ggcagttcat caaggagaag ctcatcctgc ctcacgtgga tgttcagctc 1201 aagtattttg accttgggct tccaaaccgt gaccagacca atgaccaggt caccattgac 1261 tctgctctgg ccacccagaa gtacagtgtg gctgtcaagt gtgccacaat cacccctgat 1321 gaggcccgtg tggaagagtt caagctgaag aaaatgtgga agagccctaa cggaacgatc 1381 cggaacatcc ttgggggaac cgtcttcaga gagccaatca tctgcaaaaa catcccccgc 1441 cttgtccctg gctggaccaa gcccatcacc attggcaggc acgcccatgg cgaccagtac 1501 aaggccacag attttgtggt agatcgagct ggcacgttca agttggtctt caccccaaag 1561 gatggcagca gtgccaagga gtgggaggtg tataacttcc ctgccggagg cgttgggatg 1621 gggatgtaca acaccgacga gtccatttcg ggcttcgcgc acagctgctt ccagtactct 1681 atccagaaga aatggccgct ctacttgagc accaagaaca ccattctgaa ggcctatgac 1741 gggcgtttca aggacatctt ccaggagatc tttgacaagc actataagac tgacttcgac 1801 aagaataaga tctggtatga acatcggctc atcgacgaca tggtggccca ggtgctcaag 1861 tcttccggtg gctttgtgtg ggcttgcaaa aactatgatg gagacgtgca gtctgacatc 1921 ctggctcaag gctttggctc cctcggcctg atgacatctg tgctggtctg ccctgatggg 1981 aagacaattg aggctgaggc tgctcatggg acagtcaccc gccattaccg agaacaccag 2041 aagggccggc ccaccagtac caaccctatt gccagcatct ttgcctggac acggggtctg 2101 gagcatcgtg ggaagctgga tgggaaccag gaccttatca ggtttgcaca gacgctggag 2161 aaggtgtgcg tgcagactgt ggagagcgga gcgatgacca aggacctggc tggctgtatc 2221 catggcctca gcaatgtgaa gctgaacgag cacttcctga acaccacaga cttcctggac 2281 accattaaga gcaacctgga cagagctctg ggcaagcagc tcgaggacta caaagaccat 2341 gacggtgatt ataaagatca tgacatcgac tacaaggatg acgatgacaa gtagtgaggg 2401 cccgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt 2461 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa 2521 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg 2581 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg 2641 gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac 2701 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct 2761 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg 2821 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt 2881 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca 2941 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga 3001 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa 3061 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac 3121 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag 3181 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc 3241 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag 3301 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc 3361 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc 3421 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg 3481 gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga 3541 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct 3601 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc 3661 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg 3721 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg 3781 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc 3841 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc 3901 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg 3961 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc 4021 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg 4081 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc 4141 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag 4201 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg 4261 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg 4321 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc 4381 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt 4441 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca 4501 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt 4561 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt 4621 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata 4681 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag 4741 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg 4801 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca 4861 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc 4921 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg 4981 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa 5041 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga 5101 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag 5161 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct 5221 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg 5281 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc 5341 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt 5401 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta 5461 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac 5521 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc 5581 ttgatccggc aaacaaacca ccgctggtag cggttttttt gtttgcaagc agcagattac 5641 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca 5701 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac 5761 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac 5821 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt 5881 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt 5941 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt 6001 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc 6061 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa 6121 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg 6181 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt 6241 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc 6301 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt 6361 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg 6421 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac 6481 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc 6541 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt 6601 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg 6661 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag 6721 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa 6781 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtc //