LOCUS V021923 10124 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V021923 VERSION V021923 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" regulatory 2809..2818 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 2818..2883 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" CDS 2890..4815 /label="Trim47(NM_001109585)" /note="Trim47(NM_001109585)" /gene="Trim47" misc_feature 4827..5413 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" /label="IRES2" CDS 5433..6032 /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 6046..6634 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6841..7474 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(7602..7618) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7626..7642 /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(7650..7680) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7695..7716 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(8004..8592) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8763..9623) /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(9624..9728) /gene="bla" /label="AmpR promoter" polyA_signal 9776..9910 /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 tctatttccg gtgaattcgc caccatggac 2821 tacaaagacc atgacggtga ttataaagat catgacatcg actacaagga tgacgatgac 2881 aagctcgaga tggacggcag cgggcctttc agctgcccca tctgcctgga accgctccgg 2941 gagccggtga cactaccctg cggccacaac ttctgtctcg cttgcctggg cgcgctctgg 3001 ccgcaccgga gcgcgggcgg caccggtggt tccggaggtc cggctcgctg cccactttgc 3061 caggagcctt ttcccgacgg cctgcagctc cgcaagaacc acacgctgtc cgagttgctg 3121 cagctccgcc agggctcgag ccccggaccc atgtccgccc cggctccagg ctcggcccgg 3181 ggcgcgacgc ccgagccctc cgcacccagc gcaccacctc cggctccgga gccctccgct 3241 ccctgcgcgc ccgagccgtg gcccgcgggt gaggagccgg tgcgctgcga cgcgtgcccc 3301 gagggcgccg ccctgcccgc cgcgctctcc tgcctctcct gcctcgcctc cttctgctcc 3361 gcgcacctgg ccccgcacga acgcagcccc gccctgcgcg gccaccgcct ggtgcccccg 3421 ctgcgccggc tggaggagag cctgtgcccg cgccacctgc ggccgctgga gcgctactgc 3481 cgcgtggagc gtgtgtgcct ttgcgaggcc tgcgccactc aggagcatcg gggccacgag 3541 ctcgtgccgc tggagcagga gcgcgcgctt caggaggttg aacagtccaa agtcctgagc 3601 gctgtggagg accgcatgga tgagctgggt gccggcattg cacaatcccg ccgtactgtg 3661 gccctcatca agagtgcagc tgtggcagag cgggagaggg tgagccagat gtttgccgag 3721 gccacagcct ccctacagag tttccagact gaggtgctgg ggttcatcga ggagggggag 3781 gccgccatgt tgggccgctc tcaaggtgac ctgcgcagac aagaggaaca gcgcagccgc 3841 ctaagccggg ctcgccacaa cctcagtcag gttccagagg cggattcagt cagctttctc 3901 caggagctcc tggcactgcg gctggccctg gaggaggggt gtggccctgg gcctggaccc 3961 cccagggaac tcagcttcac caagtcctcc caagcggtcc gagccgtgag ggacatcctt 4021 gtctcagcct gtgccagcca gtgggagcag ctgcgggggc tgggcagtgg tgaggatgag 4081 ctacagaaac ttggctcaga agctgatgtg gagtcccagg accctgacag caccaacctg 4141 ctggagagcg aggctcccag ggattatttc ctcaagtttg cttacattgt ggacttggac 4201 agcgacaccg cggacaagtt cctgcagctt tttggaacta aaggtgtcaa gagagtactg 4261 tgtcccatca actacccgga gtcacccacc cgattcacac actgcgaaca ggtgctagga 4321 gaaggtgccc tggaccgggg cacctactac tgggaggtgg agatcatcga aggatgggtc 4381 agtgtgggtg tcatggccga gggcttctcc ccgcaagagc cctatgaccg gggccggctg 4441 ggccgaaacg cacactcatg ctgtctgcag tggaatgggc gtgggtactc agtctggttc 4501 tgcgggctgg aagccccact accccatgcc ttttcgccta cggttggggt ctgcctggag 4561 tacgctgacc atgccctggc cttctacgct gttcgagagg ggaagctgag ccttcttcgg 4621 aggctcaaag cctcccggcc tcgccgcagc ggtgccctgg cctcccccac tgaccctttc 4681 cagagtcgcc tggacagcca tttttcaggg ctcttcaact acaggctcaa gcctgccttc 4741 ttcctggaga gtgtggatgc ccacctgcag attggacccc tcaagaaatc gtgcatatct 4801 gtgctaaaga ggaggtgagg atcccgcccc tctccctccc ccccccctaa cgttactggc 4861 cgaagccgct tggaataagg ccggtgtgcg tttgtctata tgttattttc caccatattg 4921 ccgtcttttg gcaatgtgag ggcccggaaa cctggccctg tcttcttgac gagcattcct 4981 aggggtcttt cccctctcgc caaaggaatg caaggtctgt tgaatgtcgt gaaggaagca 5041 gttcctctgg aagcttcttg aagacaaaca acgtctgtag cgaccctttg caggcagcgg 5101 aaccccccac ctggcgacag gtgcctctgc ggccaaaagc cacgtgtata agatacacct 5161 gcaaaggcgg cacaacccca gtgccacgtt gtgagttgga tagttgtgga aagagtcaaa 5221 tggctctcct caagcgtatt caacaagggg ctgaaggatg cccagaaggt accccattgt 5281 atgggatctg atctggggcc tcggtgcaca tgctttacat gtgtttagtc gaggttaaaa 5341 aaacgtctag gccccccgaa ccacggggac gtggttttcc tttgaaaaac acgatgataa 5401 gcttgccaca acccacaagg agacgacctt ccatgaccga gtacaagccc acggtgcgcc 5461 tcgccacccg cgacgacgtc ccccgggccg tacgcaccct cgccgccgcg ttcgccgact 5521 accccgccac gcgccacacc gtcgacccgg accgccacat cgagcgggtc accgagctgc 5581 aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg 5641 gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg 5701 agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg 5761 aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg 5821 tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg 5881 cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct 5941 tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca 6001 cctggtgcat gacccgcaag cccggtgcct agacgcgtct ggaacaatca acctctggat 6061 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt 6121 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc 6181 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg 6241 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc 6301 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa 6361 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat 6421 tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg tgttgccacc 6481 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt 6541 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag 6601 acgagtcgga tctccctttg ggccgcctcc ccgcctggaa ttaattctgc agtcgagacc 6661 tagaaaaaca tggagcaatc acaagtagca atacagcagc taccaatgct gattgtgcct 6721 ggctagaagc acaagaggag gaggaggtgg gttttccagt cacacctcag gtacctttaa 6781 gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa aagaggggac 6841 tggaagggct aattcactcc caacgaagac aagatatcct tgatctgtgg atctaccaca 6901 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac 6961 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca 7021 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg 7081 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag 7141 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg 7201 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat 7261 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga 7321 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct 7381 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc 7441 agaccctttt agtcagtgtg gaaaatctct agcagtagta gttcatgtca tcttattatt 7501 cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggccttg acattgctag 7561 cgtttaccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg 7621 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa 7681 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct 7741 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga 7801 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc 7861 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa 7921 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt 7981 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa 8041 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt 8101 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg 8161 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc 8221 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc 8281 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta 8341 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct 8401 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc 8461 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa 8521 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa 8581 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa 8641 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt 8701 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac 8761 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc 8821 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc 8881 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata 8941 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc 9001 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc 9061 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca 9121 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa 9181 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca 9241 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt 9301 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt 9361 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg 9421 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga 9481 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc 9541 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg 9601 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag 9661 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg 9721 gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg agatcaactt 9781 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa 9841 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca 9901 tgtctggatc aactggataa ctcaagctaa ccaaaatcat cccaaacttc ccaccccata 9961 ccctattacc actgccaatt acctgtggtt tcatttactc taaacctgtg attcctctga 10021 attattttca ttttaaagaa attgtatttg ttaaatatgt actacaaact tagtagtttt 10081 taaagaaatt gtatttgtta aatatgtact acaaacttag tagt //