LOCUS V025946 6653 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V025946 VERSION V025946 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..2143 /label="TNFRSF11B(NM_002546)" /note="TNFRSF11B(NM_002546)" /gene="TNFRSF11B" CDS 2150..2215 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 2253..2477 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2523..2951 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2965..3294 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3361..4155 /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 4329..4450 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4499..4515) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4523..4539 /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(4547..4577) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4592..4613 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4901..5486) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5657..6517) /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(6518..6622) /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 atgaacaact tgctgtgctg 961 cgcgctcgtg tttctggaca tctccattaa gtggaccacc caggaaacgt ttcctccaaa 1021 gtaccttcat tatgacgaag aaacctctca tcagctgttg tgtgacaaat gtcctcctgg 1081 tacctaccta aaacaacact gtacagcaaa gtggaagacc gtgtgcgccc cttgccctga 1141 ccactactac acagacagct ggcacaccag tgacgagtgt ctatactgca gccccgtgtg 1201 caaggagctg cagtacgtca agcaggagtg caatcgcacc cacaaccgcg tgtgcgaatg 1261 caaggaaggg cgctaccttg agatagagtt ctgcttgaaa cataggagct gccctcctgg 1321 atttggagtg gtgcaagctg gaaccccaga gcgaaataca gtttgcaaaa gatgtccaga 1381 tgggttcttc tcaaatgaga cgtcatctaa agcaccctgt agaaaacaca caaattgcag 1441 tgtctttggt ctcctgctaa ctcagaaagg aaatgcaaca cacgacaaca tatgttccgg 1501 aaacagtgaa tcaactcaaa aatgtggaat agatgttacc ctgtgtgagg aggcattctt 1561 caggtttgct gttcctacaa agtttacgcc taactggctt agtgtcttgg tagacaattt 1621 gcctggcacc aaagtaaacg cagagagtgt agagaggata aaacggcaac acagctcaca 1681 agaacagact ttccagctgc tgaagttatg gaaacatcaa aacaaagacc aagatatagt 1741 caagaagatc atccaagata ttgacctctg tgaaaacagc gtgcagcggc acattggaca 1801 tgctaacctc accttcgagc agcttcgtag cttgatggaa agcttaccgg gaaagaaagt 1861 gggagcagaa gacattgaaa aaacaataaa ggcatgcaaa cccagtgacc agatcctgaa 1921 gctgctcagt ttgtggcgaa taaaaaatgg cgaccaagac accttgaagg gcctaatgca 1981 cgcactaaag cactcaaaga cgtaccactt tcccaaaact gtcactcaga gtctaaagaa 2041 gaccatcagg ttccttcaca gcttcacaat gtacaaattg tatcagaagt tatttttaga 2101 aatgataggt aaccaggtcc aatcagtaaa aataagctgc ttactcgagg actacaaaga 2161 ccatgacggt gattataaag atcatgacat cgactacaag gatgacgatg acaagtagtg 2221 agggcccgtt taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt 2281 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc 2341 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg 2401 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga 2461 tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc 2521 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac 2581 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc 2641 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt 2701 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg 2761 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag 2821 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt 2881 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt 2941 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc 3001 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag 3061 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc 3121 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct 3181 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct 3241 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc 3301 ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc 3361 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc 3421 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca 3481 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg 3541 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg 3601 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag 3661 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg 3721 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc 3781 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa 3841 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac 3901 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat 3961 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac 4021 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc 4081 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt 4141 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc 4201 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg 4261 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg 4321 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa 4381 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca 4441 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt 4501 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg 4561 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt 4621 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg 4681 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg 4741 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa 4801 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc 4861 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc 4921 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat 4981 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc 5041 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct 5101 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg 5161 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc 5221 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga 5281 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa 5341 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta 5401 gctcttgatc cggcaaacaa accaccgctg gtagcggttt ttttgtttgc aagcagcaga 5461 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg 5521 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct 5581 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt 5641 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc 5701 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg 5761 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag 5821 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt 5881 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag 5941 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt 6001 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca 6061 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg 6121 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat 6181 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta 6241 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca 6301 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct 6361 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat 6421 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa 6481 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt 6541 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa 6601 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtc //