LOCUS V018529 7586 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V018529 VERSION V018529 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..3076 /label="USP44(NM_001042403)" /note="USP44(NM_001042403)" /gene="USP44" CDS 3083..3148 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 3186..3410 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 3456..3884 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3898..4227 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4294..5088 /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 5262..5383 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5432..5448) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5456..5472 /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(5480..5510) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5525..5546 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5834..6419) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6590..7450) /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(7451..7555) /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 atgctagcaa tggatacgtg 961 caaacatgtt gggcagctgc agcttgctca agaccattcc agcctcaacc ctcagaaatg 1021 gcactgtgtg gactgcaaca cgaccgagtc catttgggct tgccttagct gctcccatgt 1081 tgcctgtgga agatatattg aagagcatgc actcaagcac tttcaagaaa gcagtcatcc 1141 tgttgcattg gaggtgaatg agatgtacgt tttttgttac ctttgtgatg attatgttct 1201 gaatgataac acaactggag acctgaagtt actacgacgt acattaagtg ccatcaaaag 1261 tcaaaattat cactgcacaa ctcgtagtgg gaggttttta cggtccatgg gtacaggtga 1321 tgattcttat ttcttacatg acggtgccca atctctgctt caaagtgaag atcaactgta 1381 tactgctctt tggcacagga gaaggatact aatgggtaaa atctttcgaa catggtttga 1441 acaatcaccc attggaagaa aaaagcaaga agaaccattt caggaaaaaa tagtagtaaa 1501 aagagaagta aagaaaagac ggcaggaatt ggagtatcaa gttaaagcag aattggaaag 1561 tatgcctcca agaaagagtt tacgtttaca agggctcgct cagtcgacca taatagaaat 1621 agtttctgtt caggtgccag cacaaacgcc agcatcacca gcaaaagata aagtactctc 1681 tacctcagaa aatgaaatat ctcaaaaagt cagtgactcc tcagttaaac gaaggccaat 1741 agtaactcct ggtgtaacag gattgagaaa tttgggaaat acttgctata tgaattctgt 1801 tcttcaggtg ttgagtcatt tacttatttt tcgacaatgt tttttaaagc ttgatctgaa 1861 ccaatggctg gctatgactg ctagcgagaa gacaagatct tgtaagcatc caccagtcac 1921 agatacagta gtatatcaaa tgaatgaatg tcaggaaaaa gatacaggtt ttgtttgctc 1981 cagacaatca agtctgtcat caggactaag tggtggagca tcaaaaggta gaaagatgga 2041 acttattcag ccaaaggagc caacttcaca gtacatttct ctttgtcatg aattgcatac 2101 tttgttccaa gtcatgtggt ctggaaagtg ggcgttggtc tcaccatttg ctatgctaca 2161 ctcagtgtgg agactcattc ctgcctttcg tggttacgcc caacaagacg ctcaggaatt 2221 tctttgtgaa cttttagata aaatacaacg tgaattagag acaactggta ccagtttacc 2281 agctcttatc cccacttctc aaaggaaact catcaaacaa gttctgaatg ttgtaaataa 2341 catttttcat ggacaacttc ttagtcaggt tacatgtctt gcatgtgaca acaaatcaaa 2401 taccatagaa cctttctggg acttgtcatt ggagtttcca gaaaggtatc aatgcagtgg 2461 aaaagatatt gcttcccagc catgtctggt tactgaaatg ttggccaaat ttacagaaac 2521 tgaagcttta gaaggaaaaa tctacgtatg tgaccagtgt aactcaaagc gtagaaggtt 2581 ttcctccaaa ccagttgtac tcacagaagc ccagaaacaa cttatgatat gccacctacc 2641 tcaggttctc agactgcacc tcaaacgatt caggtggtca ggacgtaata accgagagaa 2701 gattggtgtt catgttggct ttgaggaaat cttaaacatg gagccctatt gctgcaggga 2761 gaccctgaaa tccctcagac cagaatgctt tatctatgac ttgtccgcgg tggtgatgca 2821 ccatgggaaa ggatttggct cagggcacta cactgcctac tgctataatt ctgaaggagg 2881 gttctgggta cactgcaatg attccaaact aagcatgtgc actatggatg aagtatgcaa 2941 ggctcaagct tatatcttgt tttataccca acgagttact gagaatggac attctaaact 3001 tttgcctcca gagctcctgt tggggagcca acatcccaat gaagacgctg atacctcgtc 3061 taatgaaatc cttagcctcg aggactacaa agaccatgac ggtgattata aagatcatga 3121 catcgactac aaggatgacg atgacaagta gtgagggccc gtttaaaccc gctgatcagc 3181 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt 3241 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca 3301 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga 3361 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc 3421 ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag 3481 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc 3541 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc 3601 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa 3661 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg 3721 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac 3781 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta 3841 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg 3901 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg 3961 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt 4021 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc 4081 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt 4141 atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc 4201 ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga 4261 tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca 4321 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc 4381 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc 4441 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg 4501 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg 4561 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct 4621 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct 4681 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa 4741 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa 4801 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc 4861 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt 4921 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct 4981 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc 5041 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg 5101 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg 5161 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc 5221 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt 5281 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac 5341 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt 5401 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt 5461 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg 5521 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg 5581 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc 5641 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc 5701 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata 5761 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg 5821 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct 5881 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa 5941 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc 6001 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt 6061 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg 6121 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg 6181 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct 6241 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc 6301 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg 6361 ctggtagcgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag 6421 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag 6481 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat 6541 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct 6601 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac 6661 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa 6721 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg 6781 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt 6841 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca 6901 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt 6961 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct 7021 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg 7081 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg 7141 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg 7201 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa 7261 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt 7321 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt 7381 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt 7441 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca 7501 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat 7561 ttccccgaaa agtgccacct gacgtc //