LOCUS V021588 9028 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V021588 VERSION V021588 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers LTR 1..634 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" misc_feature 681..806 /note="packaging signal of human immunodeficiency virus type 1" /label="HIV-1 Ψ" misc_feature 1303..1536 /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." /label="RRE" CDS 1721..1765 /codon_start=1 /note="recognized by the 2H10 single-chain llama nanobody" /product="antigenic peptide corresponding to amino acids 655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik et al., 2013)" /transl_table=1 /translation="KNEQELLELDKWASL" /label="gp41 peptide" misc_feature 2028..2143 /note="central polypurine tract and central termination sequence of HIV-1 (lacking the first T)" /label="cPPT/CTS" enhancer 2201..2504 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 2505..2708 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 2843..3763 /label="Ucp1(NM_009463)" /note="Ucp1(NM_009463)" /gene="Ucp1" CDS 3770..3796 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" promoter 3816..4315 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 4336..4935 /codon_start=1 /gene="pac from Streptomyces alboniger" /note="confers resistance to puromycin" /product="puromycin N-acetyltransferase" /transl_table=1 /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*" /label="PuroR" misc_feature 4949..5537 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5744..6377 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(6506..6522) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6530..6546 /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(6554..6584) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6599..6620 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6908..7496) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7667..8527) /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(8528..8632) /gene="bla" /label="AmpR promoter" polyA_signal 8680..8814 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" ORIGIN 1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca 61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac 121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca 181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg 241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag 301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg 361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat 421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga 481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct 541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc 601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag 661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg 721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga 781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg 841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg 901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct 961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat 1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca 1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc 1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag 1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc 1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg 1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc 1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc 1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct 1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa 1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca 1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt 1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt 1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta 1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt 1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct 1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga 1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccttt aaaagaaaag 2041 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac 2101 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg 2161 acagcagaga tccagtttat cgataagctt gggagttccg cgttacataa cttacggtaa 2221 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg 2281 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt 2341 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg 2401 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc 2461 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc 2521 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca 2581 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta 2641 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa 2701 gcagagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc 2761 tccatagaag acaccgactc tactagagga tcgctagcgc taccggactc agatctcgag 2821 ctcaagcttc gaattcgcca ccatggtgaa cccgacaact tccgaagtgc aacccaccat 2881 gggggtcaag atcttctcag ccggagtttc agcttgcctg gcagatatca tcaccttccc 2941 gctggacact gccaaagtcc gccttcagat ccaaggtgaa ggccaggctt ccagtaccat 3001 taggtataaa ggtgtcctag ggaccatcac caccctggca aaaacagaag gattgccgaa 3061 actgtacagc ggtctgcctg cgggcattca gaggcaaatc agctttgcct cactcaggat 3121 tggcctctac gactcagtcc aagagtactt ctcttcaggg agagaaacac ctgcctctct 3181 cggaaacaag atctcagccg gcttaatgac tggaggtgtg gcagtgttca ttgggcagcc 3241 tacagaggtc gtgaaggtca gaatgcaagc ccagagccat ctgcatggga tcaaaccccg 3301 ctacacgggg acctacaatg cttacagagt tatagccacc acagaaagct tgtcaacact 3361 ttggaaaggg acgaccccta atctaatgag aaatgtcatc atcaattgta cagagctggt 3421 aacatatgac ctcatgaagg gggcccttgt aaacaacaaa atactggcag atgacgtccc 3481 ctgccattta ctgtcagctc ttgttgccgg gttttgcacc acactcctgg cctctccagt 3541 ggatgtggta aaaacaagat tcatcaactc tctgccagga cagtacccaa gcgtaccaag 3601 ctgtgcgatg tccatgtaca ccaaggaagg accgacggcc tttttcaaag ggtttgtggc 3661 ttcttttctg cgactcgggt cctggaacgt catcatgttt gtgtgctttg aacagctgaa 3721 aaaagagctg atgaagtcca gacagacagt ggattgtacc acaggatcct acccatacga 3781 cgtaccagat tacgcttagt ctagataatt ctaccgggta ggggaggcgc ttttcccaag 3841 gcagtctgga gcatgcgctt tagcagcccc gctgggcact tggcgctaca caagtggcct 3901 ctggcctcgc acacattcca catccaccgg taggcgccaa ccggctccgt tctttggtgg 3961 ccccttcgcg ccaccttcta ctcctcccct agtcaggaag ttcccccccg ccccgcagct 4021 cgcgtcgtgc aggacgtgac aaatggaagt agcacgtctc actagtctcg tgcagatgga 4081 cagcaccgct gagcaatgga agcgggtagg cctttggggc agcggccaat agcagctttg 4141 ctccttcgct ttctgggctc agaggctggg aaggggtggg tccgggggcg ggctcagggg 4201 cgggctcagg ggcggggcgg gcgcccgaag gtcctccgga ggcccggcat tctgcacgct 4261 tcaaaagcgc acgtctgccg cgctgttctc ctcttcctca tctccgggcc tttcgacctg 4321 cagcccaagc ttaccatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac 4381 gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac 4441 accgtcgatc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg 4501 cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc 4561 tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg 4621 gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct cctggcgccg 4681 caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag 4741 ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg 4801 gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc 4861 ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg catgacccgc 4921 aagcccggtg cctgaccgcg tctggaacaa tcaacctctg gattacaaaa tttgtgaaag 4981 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat 5041 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc 5101 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg 5161 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct 5221 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct 5281 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg 5341 gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac 5401 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct 5461 gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct 5521 ttgggccgcc tccccgcctg gaattaattc tgcagtcgag acctagaaaa acatggagca 5581 atcacaagta gcaatacagc agctaccaat gctgattgtg cctggctaga agcacaagag 5641 gaggaggagg tgggttttcc agtcacacct caggtacctt taagaccaat gacttacaag 5701 gcagctgtag atcttagcca ctttttaaaa gaaaagaggg gactggaagg gctaattcac 5761 tcccaacgaa gacaagatat ccttgatctg tggatctacc acacacaagg ctacttccct 5821 gattagcaga actacacacc agggccaggg gtcagatatc cactgacctt tggatggtgc 5881 tacaagctag taccagttga gccagataag gtagaagagg ccaataaagg agagaacacc 5941 agcttgttac accctgtgag cctgcatggg atggatgacc cggagagaga agtgttagag 6001 tggaggtttg acagccgcct agcatttcat cacgtggccc gagagctgca tccggagtac 6061 ttcaagaact gctgatatcg agcttgctac aagggacttt ccgctgggga ctttccaggg 6121 aggcgtggcc tgggcgggac tggggagtgg cgagccctca gatcctgcat ataagcagct 6181 gctttttgcc tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg 6241 ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt 6301 gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt 6361 gtggaaaatc tctagcagta gtagttcatg tcatcttatt attcagtatt tataacttgc 6421 aaagaaatga atatcagaga gtgagaggcc ttgacattgc tagcgtttta ccgtcgacct 6481 ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc 6541 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat 6601 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc 6661 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg 6721 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag 6781 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag 6841 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc 6901 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc 6961 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc 7021 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt 7081 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg 7141 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat 7201 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag 7261 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt 7321 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc 7381 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta 7441 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag 7501 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga 7561 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa 7621 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa 7681 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc 7741 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga 7801 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa 7861 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt 7921 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg 7981 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc 8041 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg 8101 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag 8161 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt 8221 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt 8281 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac 8341 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac 8401 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag 8461 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa 8521 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga 8581 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc 8641 cccgaaaagt gccacctgac gtcgacggat cgggagatca acttgtttat tgcagcttat 8701 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg 8761 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatcaactgg 8821 ataactcaag ctaaccaaaa tcatcccaaa cttcccaccc cataccctat taccactgcc 8881 aattacctgt ggtttcattt actctaaacc tgtgattcct ctgaattatt ttcattttaa 8941 agaaattgta tttgttaaat atgtactaca aacttagtag tttttaaaga aattgtattt 9001 gttaaatatg tactacaaac ttagtagt //