LOCUS V032048 7729 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V032048 VERSION V032048 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers enhancer 50..429 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 430..633 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 744..770 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" CDS 774..800 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" CDS 804..830 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" CDS 837..2402 /label="TRAF6(NM_145803)" /note="TRAF6(NM_145803)" /gene="TRAF6" misc_feature 2505..3093 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 3159..3207 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 3409..3837 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3851..4180 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4247..5041 /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 5217..5338 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5387..5403) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5411..5427 /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(5435..5465) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5480..5501 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5789..6377) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6548..7408) /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(7409..7513) /gene="bla" /label="AmpR promoter" ORIGIN 1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta 61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag 121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc 181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga 241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat 301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc 361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct 421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca 481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat 541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg 601 cgtgtacggt gggaggtcta tataagcaga gctcgtttag tgaaccgtca gatcgcctgg 661 agacgccatc cacgctgttt tgacctccat agaagacacc gggaccgatc cagcctccgg 721 actctagagc tagcgccacc atgtacccat acgacgtacc tgattatgca acttatcctt 781 atgatgtacc tgattatgct agctatcctt atgatgtccc agactacgca aagcttatga 841 gtctgctaaa ctgtgaaaac agctgtggat ccagccagtc tgaaagtgac tgctgtgtgg 901 ccatggccag ctcctgtagc gctgtaacaa aagatgatag tgtgggtgga actgccagca 961 cggggaacct ctccagctca tttatggagg agatccaggg atatgatgta gagtttgacc 1021 cacccctgga aagcaagtat gaatgcccca tctgcttgat ggcattacga gaagcagtgc 1081 aaacgccatg cggccatagg ttctgcaaag cctgcatcat aaaatcaata agggatgcag 1141 gtcacaaatg tccagttgac aatgaaatac tgctggaaaa tcaactattt ccagacaatt 1201 ttgcaaaacg tgagattctt tctctgatgg tgaaatgtcc aaatgaaggt tgtttgcaca 1261 agatggaact gagacatctt gaggatcatc aagcacattg tgagtttgct cttatggatt 1321 gtccccaatg ccagcgtccc ttccaaaaat tccatattaa tattcacatt ctgaaggatt 1381 gtccaaggag acaggtttct tgtgacaact gtgctgcatc aatggcattt gaagataaag 1441 agatccatga ccagaactgt cctttggcaa atgtcatctg tgaatactgc aatactatac 1501 tcatcagaga acagatgcct aatcattatg atctagactg ccctacagcc ccaattccat 1561 gcacattcag tacttttggt tgccatgaaa agatgcagag gaatcacttg gcacgccacc 1621 tacaagagaa cacccagtca cacatgagaa tgttggccca ggctgttcat agtttgagcg 1681 ttatacccga ctctgggtat atctcagagg tccggaattt ccaggaaact attcaccagt 1741 tagagggtcg ccttgtaaga caagaccatc aaatccggga gctgactgct aaaatggaaa 1801 ctcagagtat gtatgtaagt gagctcaaac gaaccattcg aacccttgag gacaaagttg 1861 ctgaaatcga agcacagcag tgcaatggaa tttatatttg gaagattggc aactttggaa 1921 tgcatttgaa atgtcaagaa gaggagaaac ctgttgtgat tcatagccct ggattctaca 1981 ctggcaaacc cgggtacaaa ctgtgcatgc gcttgcacct tcagttaccg actgctcagc 2041 gctgtgcaaa ctatatatcc ctttttgtcc acacaatgca aggagaatat gacagccacc 2101 tcccttggcc cttccagggt acaatacgcc ttacaattct tgatcagtct gaagcacctg 2161 taaggcaaaa ccacgaagag ataatggatg ccaaaccaga gctgcttgct ttccagcgac 2221 ccacaatccc acggaaccca aaaggttttg gctatgtaac ttttatgcat ctggaagccc 2281 taagacaaag aactttcatt aaggatgaca cattattagt gcgctgtgag gtctccaccc 2341 gctttgacat gggtagcctt cggagggagg gttttcagcc acgaagtact gatgcagggg 2401 tataggaatt ctgcagatat ccagcacagt ggcggccgct cgagtctaga gggcccctta 2461 agggttcgat ccctaccggt tagtaatgag tttgatatct cgacaatcaa cctctggatt 2521 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg 2581 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct 2641 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc 2701 aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca 2761 ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac 2821 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt 2881 ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct 2941 ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc 3001 cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga 3061 cgagtcggat ctccctttgg gccgcctccc cgcctggaaa cgggggaggc taactgaaac 3121 acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa 3181 aacgcacggg tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact 3241 ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc 3301 caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag 3361 gccctgccat agcagatctg cgcagctggg gctctagggg gtatccccac gcgccctgta 3421 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca 3481 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct 3541 ttccccgtca agctctaaat cggggcatcc ctttagggtt ccgatttagt gctttacggc 3601 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat 3661 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc 3721 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgg 3781 ggatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat 3841 tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag 3901 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc 3961 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct 4021 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg 4081 actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa 4141 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat 4201 atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga 4261 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc 4321 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc 4381 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc 4441 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac 4501 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc 4561 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac 4621 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg 4681 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct 4741 cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt 4801 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg 4861 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac 4921 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg 4981 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg 5041 agcgggactc tggggttcgc gaaatgaccg accaagcgac gcccaacctg ccatcacgag 5101 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg 5161 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact 5221 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata 5281 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc 5341 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc 5401 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt 5461 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc 5521 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg 5581 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct 5641 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca 5701 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga 5761 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc 5821 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg 5881 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat 5941 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt 6001 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc 6061 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg 6121 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg 6181 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg 6241 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg 6301 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca 6361 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga 6421 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga 6481 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt 6541 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt 6601 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat 6661 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag 6721 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct 6781 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt 6841 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg 6901 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca 6961 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt 7021 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat 7081 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac 7141 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa 7201 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt 7261 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt 7321 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa 7381 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt 7441 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa 7501 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc 7561 tcccgatccc ctatggtcga ctctcagtac aatctgctct gatgccgcat agttaagcca 7621 gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc 7681 tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttagg //