LOCUS V020319 6824 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V020319 VERSION V020319 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 941..2314 /label="KAT8(NM_032188)" /note="KAT8(NM_032188)" /gene="KAT8" CDS 2321..2386 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 2424..2648 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2694..3122 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3136..3465 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3532..4326 /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 4500..4621 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4670..4686) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4694..4710 /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(4718..4748) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4763..4784 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5072..5657) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5828..6688) /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(6689..6793) /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 atccgccacc atggcggcac agggagctgc 961 tgcggcggtt gcggcgggga cttcaggggt cgcgggggag ggcgagcccg ggcccgggga 1021 gaatgcggcc gctgagggga ccgccccatc cccgggccgc gtctctccgc cgaccccggc 1081 gcgcggcgag ccggaagtca cggtggagat cggagaaacg tacctgtgcc ggcgaccgga 1141 tagcacctgg cattctgctg aagtgatcca gtctcgagtg aacgaccagg agggccgaga 1201 ggaattctat gtacactacg tgggctttaa ccggcggctg gacgagtggg tagacaagaa 1261 ccggctggcg ctgaccaaga cagtgaagga tgctgtacag aagaactcag agaagtacct 1321 gagcgagctc gcagagcagc ctgagcgcaa gatcactcgc aaccaaaagc gcaagcatga 1381 tgagatcaac catgtgcaga agacttatgc agagatggac cccaccacag cagccttgga 1441 gaaggagcat gaggcgatca ccaaggtgaa gtatgtggac aagatccaca tcgggaacta 1501 cgaaattgat gcctggtatt tctcaccatt ccccgaagac tatgggaaac agcccaagct 1561 ctggctctgc gagtactgcc tcaagtacat gaaatatgag aagagctacc gcttccactt 1621 gggtcagtgc cagtggcggc agccccccgg gaaagagatc taccgcaaga gcaacatctc 1681 cgtgtacgaa gttgatggca aagaccataa gatttactgt cagaacctgt gtctgctggc 1741 caagcttttc ctggaccata agacactgta ctttgacgtg gagccgttcg tcttttacat 1801 cctgactgag gtggaccggc agggggccca cattgttggc tacttctcca aggagaagga 1861 gtccccggat ggaaacaatg tggcctgcat cctgaccttg cccccctacc aacgccgcgg 1921 ctacgggaag ttcctcatcg ctttcagtta tgagctctcc aagctggaga gcacagtcgg 1981 ctccccggag aagccactgt ctgacctggg caagctcagc taccgcagct actggtcctg 2041 ggtgctgcta gagatcctgc gggacttccg gggcacactg tccatcaagg acctcagcca 2101 gatgaccagt atcacccaaa atgacatcat cagtaccctg caatccctca atatggtcaa 2161 gtactggaag ggccagcacg tgatctgtgt cacacccaag ctggtggagg agcacctcaa 2221 aagtgcccag tataagaaac cacccatcac agtggactcc gtctgcctca agtgggcacc 2281 ccccaagcac aagcaagtca agctctccaa gaagctcgag gactacaaag accatgacgg 2341 tgattataaa gatcatgaca tcgactacaa ggatgacgat gacaagtagt gagggcccgt 2401 ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc 2461 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa 2521 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg 2581 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg 2641 ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc 2701 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact 2761 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc 2821 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt 2881 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc 2941 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt 3001 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat 3061 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa 3121 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc 3181 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc 3241 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg 3301 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat 3361 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc 3421 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct 3481 tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa 3541 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac 3601 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg 3661 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag 3721 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt 3781 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg 3841 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg 3901 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga 3961 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag 4021 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat 4081 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt 4141 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg 4201 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt 4261 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc 4321 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac 4381 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg 4441 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca 4501 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa 4561 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt 4621 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt 4681 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa 4741 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac 4801 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg 4861 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc 4921 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat 4981 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca 5041 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc 5101 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc 5161 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg 5221 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta 5281 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg 5341 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac 5401 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag 5461 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat 5521 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat 5581 ccggcaaaca aaccaccgct ggtagcggtt tttttgtttg caagcagcag attacgcgca 5641 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga 5701 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga 5761 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt 5821 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt 5881 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat 5941 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag 6001 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct 6061 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt 6121 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg 6181 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca 6241 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt 6301 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat 6361 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac 6421 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa 6481 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt 6541 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt 6601 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa 6661 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt 6721 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa 6781 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtc //