LOCUS V026090 5815 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V026090 VERSION V026090 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..1287 /label="RBX1(NM_014248)" /note="RBX1(NM_014248)" /gene="RBX1" CDS 1312..1377 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 1415..1639 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 1685..2113 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2127..2456 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 2523..3317 /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 3491..3612 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(3661..3677) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 3685..3701 /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(3709..3739) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 3754..3775 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4063..4648) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(4819..5679) /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(5680..5784) /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 accatggcgg cagcgatgga tgtggatacc ccgagcggca ccaacagcgg cgcgggcaag 1021 aagcgctttg aagtgaaaaa gtggaatgca gtagccctct gggcctggga tattgtggtt 1081 gataactgtg ccatctgcag gaaccacatt atggatcttt gcatagaatg tcaagctaac 1141 caggcgtccg ctacttcaga agagtgtact gtcgcatggg gagtctgtaa ccatgctttt 1201 cacttccact gcatctctcg ctggctcaaa acacgacagg tgtgtccatt ggacaacaga 1261 gagtgggaat tccaaaagta tgggcacggt agcggcagcg gtagcctcga ggactacaaa 1321 gaccatgacg gtgattataa agatcatgac atcgactaca aggatgacga tgacaagtag 1381 tgagggcccg tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct 1441 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt 1501 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg 1561 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg 1621 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat 1681 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg 1741 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc 1801 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga 1861 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt 1921 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat 1981 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat 2041 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa 2101 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct 2161 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa 2221 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa 2281 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt 2341 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct 2401 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc 2461 tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc 2521 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat 2581 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt 2641 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac 2701 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg 2761 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc 2821 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa 2881 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc 2941 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg 3001 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg 3061 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa 3121 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg 3181 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct 3241 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc 3301 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa 3361 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat 3421 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt 3481 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac 3541 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat 3601 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg 3661 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc 3721 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc 3781 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat 3841 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac 3901 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt 3961 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca 4021 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc 4081 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact 4141 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct 4201 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag 4261 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca 4321 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa 4381 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc 4441 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag 4501 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg 4561 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ttttttgttt gcaagcagca 4621 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga 4681 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat 4741 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga 4801 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg 4861 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga 4921 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc 4981 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac 5041 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc 5101 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc 5161 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc 5221 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt 5281 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc 5341 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg 5401 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag 5461 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat 5521 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc 5581 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa 5641 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta 5701 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa 5761 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtc //