LOCUS V046143 8938 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V046143 VERSION V046143 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..3634 /label="Clec9a(NM_001205363)" /note="Clec9a(NM_001205363)" /gene="Clec9a" CDS 3641..3706 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3726..4225 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 4246..4845 /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 4859..5447 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5654..6287 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(6416..6432) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6440..6456 /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(6464..6494) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6509..6530 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6818..7406) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7577..8437) /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(8438..8542) /gene="bla" /label="AmpR promoter" polyA_signal 8590..8724 /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 ccatgcatgc ggaagaaata tatacctctc ttcagtggga 2881 cattcctacc tcagaggcct ctcagaagtg ccaatcccct agcaaatgtt caggagcatg 2941 gtgtgttgtg acgatgattt cctgtgtggt ctgtatgggc ttgttagcaa cgtccatttt 3001 cttgggcatc aagttcttcc aggtatcctc tcttgtcttg gagcagcagg aaagactcat 3061 ccaacaggac acagcattgg tgaaccttac acagtggcag aggaaataca cactggaata 3121 ctgccaagcc ttactgcaga gatctctcca ttcaggcaca gatgcttcta ctggaccagt 3181 tcttctgacc tctccacaga tggttccaca gaccctggac agcaaggaaa caggtagtga 3241 ctgcagccct tgtccacaca actggattca gaatggaaaa agttgttact atgtctttga 3301 acgctgggaa atgtggaaca tcagtaagaa gagctgttta aaagagggcg ctagtctctt 3361 tcaaatagac agcaaagaag aaatggagtt catcagcagt atagggaaac tcaaaggagg 3421 aaataaatat tgggtgggag tgtttcaaga tggaatcagt ggatcttggt tctgggaaga 3481 tggctcttct cctctctctg acttgttgcc agcagaaaga cagcgatcag ccggccagat 3541 ctgtggatac ctcaaagatt ctactctcat ctcagataag tgcgatagct ggaaatattt 3601 tatctgtgag aagaaggcat ttggatcctg catcggatcc gactacaaag accatgacgg 3661 tgattataaa gatcatgaca tcgactacaa ggatgacgat gacaagtagt ctagataatt 3721 ctaccgggta ggggaggcgc ttttcccaag gcagtctgga gcatgcgctt tagcagcccc 3781 gctgggcact tggcgctaca caagtggcct ctggcctcgc acacattcca catccaccgg 3841 taggcgccaa ccggctccgt tctttggtgg ccccttcgcg ccaccttcta ctcctcccct 3901 agtcaggaag ttcccccccg ccccgcagct cgcgtcgtgc aggacgtgac aaatggaagt 3961 agcacgtctc actagtctcg tgcagatgga cagcaccgct gagcaatgga agcgggtagg 4021 cctttggggc agcggccaat agcagctttg ctccttcgct ttctgggctc agaggctggg 4081 aaggggtggg tccgggggcg ggctcagggg cgggctcagg ggcggggcgg gcgcccgaag 4141 gtcctccgga ggcccggcat tctgcacgct tcaaaagcgc acgtctgccg cgctgttctc 4201 ctcttcctca tctccgggcc tttcgacctg cagcccaagc ttaccatgac cgagtacaag 4261 cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc 4321 gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg 4381 gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg 4441 gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg 4501 gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg 4561 cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg 4621 gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc 4681 cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc 4741 cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc 4801 gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgaccgcg tctggaacaa 4861 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc 4921 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat 4981 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg 5041 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg 5101 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat 5161 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt 5221 gggcactgac aattccgtgg tgttgtcggg gaagctgacg tcctttccat ggctgctcgc 5281 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa 5341 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg 5401 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg gaattaattc 5461 tgcagtcgag acctagaaaa acatggagca atcacaagta gcaatacagc agctaccaat 5521 gctgattgtg cctggctaga agcacaagag gaggaggagg tgggttttcc agtcacacct 5581 caggtacctt taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa 5641 gaaaagaggg gactggaagg gctaattcac tcccaacgaa gacaagatat ccttgatctg 5701 tggatctacc acacacaagg ctacttccct gattagcaga actacacacc agggccaggg 5761 gtcagatatc cactgacctt tggatggtgc tacaagctag taccagttga gccagataag 5821 gtagaagagg ccaataaagg agagaacacc agcttgttac accctgtgag cctgcatggg 5881 atggatgacc cggagagaga agtgttagag tggaggtttg acagccgcct agcatttcat 5941 cacgtggccc gagagctgca tccggagtac ttcaagaact gctgatatcg agcttgctac 6001 aagggacttt ccgctgggga ctttccaggg aggcgtggcc tgggcgggac tggggagtgg 6061 cgagccctca gatcctgcat ataagcagct gctttttgcc tgtactgggt ctctctggtt 6121 agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca 6181 ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa 6241 ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta gtagttcatg 6301 tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga gtgagaggcc 6361 ttgacattgc tagcgtttta ccgtcgacct ctagctagag cttggcgtaa tcatggtcat 6421 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa 6481 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc 6541 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc 6601 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact 6661 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac 6721 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa 6781 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg 6841 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa 6901 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc 6961 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac 7021 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac 7081 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg 7141 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt 7201 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa 7261 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct 7321 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga 7381 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg 7441 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct 7501 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt 7561 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc 7621 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg 7681 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag 7741 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt 7801 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag 7861 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt 7921 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca 7981 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg 8041 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat 8101 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta 8161 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca 8221 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct 8281 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat 8341 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa 8401 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt 8461 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa 8521 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat 8581 cgggagatca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca 8641 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc 8701 aatgtatctt atcatgtctg gatcaactgg ataactcaag ctaaccaaaa tcatcccaaa 8761 cttcccaccc cataccctat taccactgcc aattacctgt ggtttcattt actctaaacc 8821 tgtgattcct ctgaattatt ttcattttaa agaaattgta tttgttaaat atgtactaca 8881 aacttagtag tttttaaaga aattgtattt gttaaatatg tactacaaac ttagtagt //