LOCUS V032845 7777 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V032845 VERSION V032845 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 964..3267 /label="CUL3(NM_003590)" /note="CUL3(NM_003590)" /gene="CUL3" CDS 3274..3339 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 3377..3601 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 3647..4075 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 4089..4418 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4485..5279 /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 5453..5574 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5623..5639) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5647..5663 /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(5671..5701) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5716..5737 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6025..6610) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6781..7641) /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(7642..7746) /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 atccactagt ccagtgtggt ggaattcgcc 961 accatgtcga atctgagcaa aggcacgggc agccggaagg acaccaagat gcggatccgg 1021 gcctttccga tgaccatgga tgaaaaatat gtaaacagca tttgggacct tctgaaaaat 1081 gcaattcaag aaatccagcg taagaataac agtggtctta gttttgagga gctctataga 1141 aatgcatata caatggtttt gcataaacat ggagaaaagc tctacactgg actaagagaa 1201 gttgttaccg aacatctcat aaataaggtg cgagaagatg tactaaattc attgaataac 1261 aactttcttc aaacgctaaa tcaagcttgg aatgatcatc aaacagctat ggtgatgatt 1321 agagacatac taatgtacat ggaccgtgtg tatgtacaac aaaataatgt ggagaacgtc 1381 tacaatttgg gattaattat ttttcgagat caagttgtac gttatgggtg tattagggat 1441 catctacggc aaactctatt ggatatgatt gcaagagagc ggaaaggaga agtcgtagac 1501 agaggcgcaa taagaaatgc ttgccagatg ttaatgattt taggtctcga aggaagatca 1561 gtctatgaag aagattttga ggctcctttt ttggaaatgt ctgcagaatt ttttcagatg 1621 gaaagccaga aatttttagc agaaaatagt gcttcagtat atataaagaa agtagaagct 1681 agaattaatg aagaaataga acgagtgatg cactgccttg acaaatcaac ggaagaacca 1741 attgtaaagg tggttgaaag ggaactcatt tccaagcaca tgaagactat agtagaaatg 1801 gagaattctg ggctagtaca tatgttgaaa aatggaaaga cagaagacct tggttgcatg 1861 tacaagttat ttagtcgtgt gccaaatggt ttgaaaacaa tgtgtgagtg tatgagttcc 1921 tatttgaggg agcaaggtaa agctcttgtt tctgaagaag gagaaggaaa gaatcctgtt 1981 gactatatcc agggcttatt ggatctgaag agtaggttcg atcgcttcct cctggaatca 2041 ttcaacaatg accgtctctt taaacaaact attgcgggtg actttgagta ttttctcaac 2101 ctcaactcca ggtctcctga atacctctca ttatttattg atgataagct gaaaaaggga 2161 gtcaaagggc taacagaaca agaagtagaa acaatattgg ataaagcaat ggtccttttt 2221 aggtttatgc aagaaaaaga tgtatttgaa cgttattata aacaacactt ggcaaggaga 2281 cttctcacaa ataaaagtgt ttctgatgac tctgaaaaaa acatgatatc taagttaaag 2341 actgaatgtg gatgtcagtt cacgtcaaaa ctggaaggaa tgtttaggga tatgagcatc 2401 tcaaacacaa cgatggatga attcaggcaa catctacagg caactggtgt atctttaggt 2461 ggtgttgatc ttacagtccg ggtgctcacg acaggatatt ggcccactca gtcagccaca 2521 ccaaagtgca acatcccacc agcaccaaga catgcttttg agatattcag aaggttctac 2581 ttagccaaac acagtggtcg acagctcaca ctccagcatc atatgggttc tgcagatctc 2641 aatgccacat tttatggacc agttaaaaag gaagatggat ctgaagttgg tgttggaggt 2701 gcacaagtaa ctggctctaa tacacggaag cacatattgc aagtttccac tttccagatg 2761 accatattaa tgctctttaa taatagagaa aaatacacat ttgaggaaat tcagcaagag 2821 acagatatcc ctgaaagaga gcttgttaga gccctacagt ccctcgcctg tggtaaacca 2881 acacagcggg ttcttacaaa agaacccaaa tcaaaggaaa tagaaaatgg tcatatattt 2941 acagttaatg atcaattcac atccaaacta cacagagtca agattcaaac agttgctgcc 3001 aaacaaggtg aatccgaccc agagaggaaa gaaacaaggc agaaagtaga cgacgacaga 3061 aaacatgaga tagaagctgc tatagtgcgg ataatgaaat ctagaaagaa gatgcagcac 3121 aatgttctag tagcggaggt aactcagcag ttgaaggcgc gattcttacc aagtccagtt 3181 gttattaaga aacgtattga aggacttatt gagagagaat atttggcacg aacacctgag 3241 gatcgcaaag tatacacata tgtagcactc gaggactaca aagaccatga cggtgattat 3301 aaagatcatg acatcgacta caaggatgac gatgacaagt agtgagggcc cgtttaaacc 3361 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc 3421 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa 3481 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac 3541 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg 3601 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc 3661 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc 3721 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt 3781 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac 3841 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag 3901 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa 3961 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg 4021 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc 4081 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta 4141 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag 4201 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa 4261 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac 4321 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt 4381 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat 4441 ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg 4501 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac 4561 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg 4621 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc 4681 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg 4741 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc 4801 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc 4861 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta 4921 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg 4981 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg 5041 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat 5101 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc 5161 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta 5221 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag 5281 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt 5341 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg 5401 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt 5461 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc 5521 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt 5581 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt 5641 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa 5701 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc 5761 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag 5821 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt 5881 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga 5941 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg 6001 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa 6061 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt 6121 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct 6181 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct 6241 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc 6301 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt 6361 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc 6421 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat 6481 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa 6541 acaaaccacc gctggtagcg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa 6601 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa 6661 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt 6721 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag 6781 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat 6841 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc 6901 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa 6961 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca 7021 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa 7081 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt 7141 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc 7201 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact 7261 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc 7321 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg 7381 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct 7441 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc 7501 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag 7561 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac 7621 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg 7681 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt 7741 tccgcgcaca tttccccgaa aagtgccacc tgacgtc //