LOCUS V022725 6785 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V022725 VERSION V022725 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 908..2263 /label="IDH2(NM_002168)" /note="IDH2(NM_002168)" /gene="IDH2" CDS 2264..2305 /codon_start=1 /product="epitope tag from simian virus 5" /transl_table=1 /translation="GKPIPNPLLGLDST" /label="V5 tag" polyA_signal 2352..2576 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2622..3050 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3064..3393 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3460..4254 /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 4428..4549 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4598..4614) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4622..4638 /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(4646..4676) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4691..4712 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5000..5585) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5756..6616) /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(6617..6721) /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 cgccaccatg gccggctacc tgcgggtcgt gcgctcgctc tgcagagcct caggctcgcg 961 gccggcctgg gcgccggcgg ccctgacagc ccccacctcg caagagcagc cgcggcgcca 1021 ctatgccgac aaaaggatca aggtggcgaa gcccgtggtg gagatggatg gtgatgagat 1081 gacccgtatt atctggcagt tcatcaagga gaagctcatc ctgccccacg tggacatcca 1141 gctaaagtat tttgacctcg ggctcccaaa ccgtgaccag actgatgacc aggtcaccat 1201 tgactctgca ctggccaccc agaagtacag tgtggctgtc aagtgtgcca ccatcacccc 1261 tgatgaggcc cgtgtggaag agttcaagct gaagaagatg tggaaaagtc ccaatggaac 1321 tatccggaac atcctggggg ggactgtctt ccgggagccc atcatctgca aaaacatccc 1381 acgcctagtc cctggctgga ccaagcccat caccattggc aggcacgccc atggcgacca 1441 gtacaaggcc acagactttg tggcagaccg ggccggcact ttcaaaatgg tcttcacccc 1501 aaaagatggc agtggtgtca aggagtggga agtgtacaac ttccccgcag gcggcgtggg 1561 catgggcatg tacaacaccg acgagtccat ctcaggtttt gcgcacagct gcttccagta 1621 tgccatccag aagaaatggc cgctgtacat gagcaccaag aacaccatac tgaaagccta 1681 cgatgggcgt ttcaaggaca tcttccagga gatctttgac aagcactata agaccgactt 1741 cgacaagaat aagatctggt atgagcaccg gctcattgat gacatggtgg ctcaggtcct 1801 caagtcttcg ggtggctttg tgtgggcctg caagaactat gacggagatg tgcagtcaga 1861 catcctggcc cagggctttg gctcccttgg cctgatgacg tccgtcctgg tctgccctga 1921 tgggaagacg attgaggctg aggccgctca tgggaccgtc acccgccact atcgggagca 1981 ccagaagggc cggcccacca gcaccaaccc catcgccagc atctttgcct ggacacgtgg 2041 cctggagcac cgggggaagc tggatgggaa ccaagacctc atcaggtttg cccagatgct 2101 ggagaaggtg tgcgtggaga cggtggagag tggagccatg accaaggacc tggcgggctg 2161 cattcacggc ctcagcaatg tgaagctgaa cgagcacttc ctgaacacca cggacttcct 2221 cgacaccatc aagagcaacc tggacagagc cctgggcagg cagggtaagc ctatccctaa 2281 ccctctcctc ggtctcgatt ctacgtgact cgagtctaga gggcccgttt aaacccgctg 2341 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc 2401 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc 2461 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa 2521 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc 2581 tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc 2641 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct 2701 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg 2761 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga 2821 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt 2881 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg 2941 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc 3001 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg 3061 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa 3121 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc 3181 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg 3241 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt 3301 ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga 3361 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt 3421 ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg 3481 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag 3541 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt 3601 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta 3661 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg 3721 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt 3781 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat 3841 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg 3901 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca 3961 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc 4021 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc 4081 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat 4141 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc 4201 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga 4261 ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt 4321 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga 4381 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg 4441 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt 4501 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta 4561 taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa 4621 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct 4681 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc 4741 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg 4801 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc 4861 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag 4921 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa 4981 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc 5041 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc 5101 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg 5161 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt 5221 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc 5281 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc 5341 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag 5401 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg 5461 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa 5521 ccaccgctgg tagcggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat 5581 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac 5641 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt 5701 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc 5761 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg 5821 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg 5881 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc 5941 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta 6001 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg 6061 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct 6121 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta 6181 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg 6241 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga 6301 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt 6361 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca 6421 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt 6481 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt 6541 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga 6601 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt 6661 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc 6721 gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct 6781 atggt //