LOCUS V032339 7906 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V032339 VERSION V032339 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..2565 /label="KLHL32(NM_052904)" /note="KLHL32(NM_052904)" /gene="KLHL32" CDS 2572..2598 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" CDS 2602..2628 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" CDS 2632..2658 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" misc_feature 2682..3270 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 3336..3384 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 3586..4014 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 4028..4357 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4424..5218 /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 5394..5515 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5564..5580) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5588..5604 /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(5612..5642) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5657..5678 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5966..6554) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6725..7585) /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(7586..7690) /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 ccaccatgcc gtctgaacgc 721 tgcctcagta ttcaagaaat gctgacaggc cagaggctct gccactccga atctcacaat 781 gacagtgtcc tggcagcgct gaatcagcag aggagtgatg gcatcctctg cgacatcacc 841 ctgattgctg aggaacagaa attccatgct cacaaggcag tcctagcagc atgcagtgac 901 tatttccggg caatgttcag tctttgtatg gtggaaagtg gagctgatga ggttaatttg 961 cacggtgtga ccagccttgg cttaaagcag gctctggagt ttgcatacac aggacagatt 1021 ttgctggagc caggtgtgat ccaggatgtg ctagcagcgg gcagtcacct acagctgttg 1081 gagcttctca atttatgctc ccactatctc atccaggaat taaatagctt taattacttg 1141 gatctgtaca gacttgctga cctctttaac ctcactttgt tggagaaggc agtgatcgat 1201 ttcttagtga aacatctctc tgaactcctg aagagccgcc cagaagaagt tctaacgctt 1261 ccctattgcc tgcttcagga ggtgctgaag agcgaccgcc tgacctccct gagtgaagag 1321 cagatctggc agctagctgt gaggtggttg gaacacaact gccactacca gtacatggac 1381 gagctcctgc aatacatccg ctttggccta atggatgtgg atactctcca tacagttgcc 1441 ctgtcccacc cccttgtcca agcaagtgag actgcaacag cccttgtcaa cgaggccctg 1501 gaataccacc agagcatcta tgcacagcct gtctggcaga ctcgcaggac caaaccacga 1561 ttccagtcag acactctgta tatcattggt gggaaaaagc gcgaggtctg caaggtcaag 1621 gaacttcggt acttcaatcc tgttgatcag gagaatgctc tcatagctgc cattgccaac 1681 tggagtgagc tggctcccat gcctgtggga aggagccacc attgtgtggc agtcatgggg 1741 gacttcctgt ttgtggcagg aggggaagtt gagcatgcca gtggccggac gtgtgctgtg 1801 aggactgcct gtcgctatga cccccgcagt aattcctggg cagagatagc acccatgaaa 1861 aactgccggg agcattttgt gctgggtgcc atggaggaat acctctatgc agttgggggc 1921 agaaatgaac tgcgccaggt tctgcctaca gttgagcgat attgccccaa gaagaacaaa 1981 tggacttttg ttcagtcctt tgacagatcc ctttcatgcc atgctggata tgtggctgat 2041 ggtcttcttt ggatatcagg tggagtaact aatacggcac aatatcagaa caggctaatg 2101 gtgtatgaac ctaaccagaa taagtggata agccgtagcc ccatgctgca gagaagggtc 2161 taccattcca tggctgctgt acaaaggaag ctttatgttc ttggaggcaa tgacctagac 2221 tacaataatg accggatcct tgtgcgccat atagattctt acaacataga cactgaccag 2281 tggacacgtt gtaatttcaa cctgctgact ggccaaaatg aatctggagt tgctgtccat 2341 aatgggagaa tatatttagt tggtggatat tcaatttgga caaatgagcc tctggcttgt 2401 atccaggtac tggatgtaag cagagaaggc aaagaagaag tattctatgg gcctacactc 2461 ccttttgctt ccaatggaat agcagcatgc ttccttccag ctccatattt tacatgccct 2521 aaccttcaaa ctcttcaagt gcctcatcac aggattggca ccatcctcga gtacccatac 2581 gacgtacctg attatgcaac ttatccttat gatgtacctg attatgctag ctatccttat 2641 gatgtcccag actacgcata gtgagggccc gatatctcga caatcaacct ctggattaca 2701 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat 2761 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct 2821 ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac 2881 gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca 2941 cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca 3001 tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg 3061 tggtgttgtc ggggaagctg acgtcctttc catggctgct cgcctgtgtt gccacctgga 3121 ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt 3181 cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga 3241 gtcggatctc cctttgggcc gcctccccgc ctggaaacgg gggaggctaa ctgaaacacg 3301 gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac 3361 gcacgggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg 3421 tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac 3481 cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc 3541 ctgccatagc agatctgcgc agctggggct ctagggggta tccccacgcg ccctgtagcg 3601 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg 3661 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc 3721 cccgtcaagc tctaaatcgg ggcatccctt tagggttccg atttagtgct ttacggcacc 3781 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga 3841 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa 3901 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgggga 3961 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct 4021 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat 4081 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc 4141 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac 4201 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact 4261 aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta 4321 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc 4381 cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg 4441 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca 4501 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt 4561 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg 4621 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga 4681 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca 4741 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct 4801 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac 4861 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc 4921 gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt 4981 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt 5041 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg 5101 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat 5161 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc 5221 gggactctgg ggttcgcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt 5281 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg 5341 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt 5401 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag 5461 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg 5521 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg 5581 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta 5641 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg 5701 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga 5761 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg 5821 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag 5881 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc 5941 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca 6001 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt 6061 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc 6121 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc 6181 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc 6241 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact 6301 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg 6361 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta 6421 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca 6481 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa 6541 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg 6601 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc 6661 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg 6721 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat 6781 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg 6841 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa 6901 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca 6961 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc 7021 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt 7081 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa 7141 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat 7201 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct 7261 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga 7321 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag 7381 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga 7441 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca 7501 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg 7561 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc 7621 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag 7681 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc 7741 cgatccccta tggtcgactc tcagtacaat ctgctctgat gccgcatagt taagccagta 7801 tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac 7861 aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagg //