LOCUS V021489 7430 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V021489 VERSION V021489 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 683..2077 /label="PRKN(NM_004562)" /note="PRKN(NM_004562)" /gene="PRKN" CDS 2117..2182 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 2206..2794 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 2860..2908 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 3110..3538 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3552..3881 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3948..4742 /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 4918..5039 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5088..5104) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5112..5128 /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(5136..5166) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5181..5202 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5490..6078) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6249..7109) /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(7110..7214) /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 ggatccgcca ccatgatagt gtttgtcagg ttcaactcca gccatggttt 721 cccagtggag gtcgattctg acaccagcat cttccagctc aaggaggtgg ttgctaagcg 781 acagggggtt ccggctgacc agttgcgtgt gattttcgca gggaaggagc tgaggaatga 841 ctggactgtg cagaattgtg acctggatca gcagagcatt gttcacattg tgcagagacc 901 gtggagaaaa ggtcaagaaa tgaatgcaac tggaggcgac gaccccagaa acgcggcggg 961 aggctgtgag cgggagcccc agagcttgac tcgggtggac ctcagcagct cagtcctccc 1021 aggagactct gtggggctgg ctgtcattct gcacactgac agcaggaagg actcaccacc 1081 agctggaagt ccagcaggta gatcaatcta caacagcttt tatgtgtatt gcaaaggccc 1141 ctgtcaaaga gtgcagccgg gaaaactcag ggtacagtgc agcacctgca ggcaggcaac 1201 gctcaccttg acccagggtc catcttgctg ggatgatgtt ttaattccaa accggatgag 1261 tggtgaatgc caatccccac actgccctgg gactagtgca gaatttttct ttaaatgtgg 1321 agcacacccc acctctgaca aggaaacatc agtagctttg cacctgatcg caacaaatag 1381 tcggaacatc acttgcatta cgtgcacaga cgtcaggagc cccgtcctgg ttttccagtg 1441 caactcccgc cacgtgattt gcttagactg tttccactta tactgtgtga caagactcaa 1501 tgatcggcag tttgttcacg accctcaact tggctactcc ctgccttgtg tggctggctg 1561 tcccaactcc ttgattaaag agctccatca cttcaggatt ctgggagaag agcagtacaa 1621 ccggtaccag cagtatggtg cagaggagtg tgtcctgcag atggggggcg tgttatgccc 1681 ccgccctggc tgtggagcgg ggctgctgcc ggagcctgac cagaggaaag tcacctgcga 1741 agggggcaat ggcctgggct gtgggtttgc cttctgccgg gaatgtaaag aagcgtacca 1801 tgaaggggag tgcagtgccg tatttgaagc ctcaggaaca actactcagg cctacagagt 1861 cgatgaaaga gccgccgagc aggctcgttg ggaagcagcc tccaaagaaa ccatcaagaa 1921 aaccaccaag ccctgtcccc gctgccatgt accagtggaa aaaaatggag gctgcatgca 1981 catgaagtgt ccgcagcccc agtgcaggct cgagtggtgc tggaactgtg gctgcgagtg 2041 gaaccgcgtc tgcatggggg accactggtt cgacgtggaa ttctgcagat atccagcaca 2101 gtggcggccg ctcgaggact acaaagacca tgacggtgat tataaagatc atgacatcga 2161 ctacaaggat gacgatgaca agtagtgagg gcccgatatc tcgacaatca acctctggat 2221 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt 2281 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc 2341 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg 2401 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc 2461 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa 2521 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat 2581 tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg tgttgccacc 2641 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt 2701 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag 2761 acgagtcgga tctccctttg ggccgcctcc ccgcctggaa acgggggagg ctaactgaaa 2821 cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata 2881 aaacgcacgg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac 2941 tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc 3001 ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca 3061 ggccctgcca tagcagatct gcgcagctgg ggctctaggg ggtatcccca cgcgccctgt 3121 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc 3181 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc 3241 tttccccgtc aagctctaaa tcggggcatc cctttagggt tccgatttag tgctttacgg 3301 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga 3361 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc 3421 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg 3481 gggatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa 3541 ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa 3601 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc 3661 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc 3721 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct 3781 gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga 3841 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta 3901 tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag 3961 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg 4021 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc 4081 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag 4141 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca 4201 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat 4261 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata 4321 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac 4381 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc 4441 tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg 4501 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg 4561 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta 4621 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg 4681 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct 4741 gagcgggact ctggggttcg cgaaatgacc gaccaagcga cgcccaacct gccatcacga 4801 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac 4861 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac 4921 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat 4981 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat 5041 catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt 5101 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag 5161 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg 5221 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg 5281 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc 5341 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc 5401 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg 5461 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat 5521 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag 5581 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga 5641 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg 5701 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt 5761 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac 5821 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc 5881 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt 5941 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc 6001 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc 6061 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg 6121 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag 6181 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg 6241 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt 6301 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca 6361 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca 6421 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc 6481 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt 6541 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg 6601 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc 6661 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg 6721 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga 6781 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga 6841 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta 6901 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg 6961 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact 7021 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata 7081 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt 7141 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa 7201 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat 7261 ctcccgatcc cctatggtcg actctcagta caatctgctc tgatgccgca tagttaagcc 7321 agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag 7381 ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg //