LOCUS V025410 10114 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V025410 VERSION V025410 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 2815..4734 /label="UBA2(NM_005499)" /note="UBA2(NM_005499)" /gene="UBA2" CDS 4741..4806 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 4817..5403 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" /label="IRES2" CDS 5423..6022 /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 6036..6624 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6831..7464 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(7592..7608) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7616..7632 /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(7640..7670) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7685..7706 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7994..8582) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8753..9613) /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(9614..9718) /gene="bla" /label="AmpR promoter" polyA_signal 9766..9900 /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 caccatggca 2821 ctgtcgcggg ggctgccccg ggagctggct gaggcggtgg ccgggggccg ggtgctggtg 2881 gtgggggcgg gcggcatcgg ctgcgagctc ctcaagaatc tcgtgctcac cggtttctcc 2941 cacatcgacc tgattgatct ggatactatt gatgtaagca acctcaacag acagtttttg 3001 tttcaaaaga aacatgttgg aagatcaaag gcacaggttg ccaaggaaag tgtactgcag 3061 ttttacccga aagctaatat cgttgcctac catgacagca tcatgaaccc tgactataat 3121 gtggaatttt tccgacagtt tatactggtt atgaatgctt tagataacag agctgcccga 3181 aaccatgtta atagaatgtg cctggcagct gatgttcctc ttattgaaag tggaacagct 3241 gggtatcttg gacaagtaac tactatcaaa aagggtgtga ccgagtgtta tgagtgtcat 3301 cctaagccga cccagagaac ctttcctggc tgtacaattc gtaacacacc ttcagaacct 3361 atacattgca tcgtttgggc aaagtacttg ttcaaccagt tgtttgggga agaagatgct 3421 gatcaagaag tatctcctga cagagctgac cctgaagctg cctgggaacc aacggaagcc 3481 gaagccagag ctagagcatc taatgaagat ggtgacatta aacgtatttc tactaaggaa 3541 tgggctaaat caactggata tgatccagtt aaacttttta ccaagctttt taaagatgac 3601 atcaggtatc tgttgacaat ggacaaacta tggcggaaaa ggaaacctcc agttccgttg 3661 gactgggctg aagtacaaag tcaaggagaa gaaacgaatg catcagatca acagaatgaa 3721 ccccagttag gcctgaaaga ccagcaggtt ctagatgtaa agagctatgc acgtcttttt 3781 tcaaagagca tcgagacttt gagagttcat ttagcagaaa agggggatgg agctgagctc 3841 atatgggata aggatgaccc atctgcaatg gattttgtca cctctgctgc aaacctcagg 3901 atgcatattt tcagtatgaa tatgaagagt agatttgata tcaaatcaat ggcagggaac 3961 attattcctg ctattgctac tactaatgca gtaattgctg ggttgatagt attggaagga 4021 ttgaagattt tatcaggaaa aatagaccag tgcagaacaa tttttttgaa taaacaacca 4081 aacccaagaa agaagcttct tgtgccttgt gcactggatc ctcccaaccc caattgttat 4141 gtatgtgcca gcaagccaga ggtgactgtg cggctgaatg tccataaagt gactgttctc 4201 accttacaag acaagatagt gaaagaaaaa tttgctatgg tagcaccaga tgtccaaatt 4261 gaagatggga aaggaacaat cctaatatct tccgaagagg gagagacgga agctaataat 4321 cacaagaagt tgtcagaatt tggaattaga aatggcagcc ggcttcaagc agatgacttc 4381 ctccaggact atactttatt gatcaacatc cttcatagtg aagacctagg aaaggacgtt 4441 gaatttgaag ttgttggtga tgccccggaa aaagtggggc ccaaacaagc tgaagatgct 4501 gccaaaagca taaccaatgg cagtgatgat ggagctcagc cctccacctc cacagctcaa 4561 gagcaagatg acgttctcat agttgattca gatgaagaag attcttcaaa taatgccgac 4621 gtcagtgaag aagagagaag ccgcaagagg aaattagatg agaaagagaa tctcagtgca 4681 aagaggtcac gtatagaaca gaaggaagag cttgatgatg tcatagcatt agatctcgag 4741 gactacaaag accatgacgg tgattataaa gatcatgaca tcgactacaa ggatgacgat 4801 gacaagtagg atcccgcccc tctccctccc ccccccctaa cgttactggc cgaagccgct 4861 tggaataagg ccggtgtgcg tttgtctata tgttattttc caccatattg ccgtcttttg 4921 gcaatgtgag ggcccggaaa cctggccctg tcttcttgac gagcattcct aggggtcttt 4981 cccctctcgc caaaggaatg caaggtctgt tgaatgtcgt gaaggaagca gttcctctgg 5041 aagcttcttg aagacaaaca acgtctgtag cgaccctttg caggcagcgg aaccccccac 5101 ctggcgacag gtgcctctgc ggccaaaagc cacgtgtata agatacacct gcaaaggcgg 5161 cacaacccca gtgccacgtt gtgagttgga tagttgtgga aagagtcaaa tggctctcct 5221 caagcgtatt caacaagggg ctgaaggatg cccagaaggt accccattgt atgggatctg 5281 atctggggcc tcggtgcaca tgctttacat gtgtttagtc gaggttaaaa aaacgtctag 5341 gccccccgaa ccacggggac gtggttttcc tttgaaaaac acgatgataa gcttgccaca 5401 acccacaagg agacgacctt ccatgaccga gtacaagccc acggtgcgcc tcgccacccg 5461 cgacgacgtc ccccgggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac 5521 gcgccacacc gtcgacccgg accgccacat cgagcgggtc accgagctgc aagaactctt 5581 cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt 5641 ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc 5701 gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct 5761 ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga 5821 ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg 5881 cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg 5941 gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat 6001 gacccgcaag cccggtgcct agacgcgtct ggaacaatca acctctggat tacaaaattt 6061 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg 6121 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt 6181 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg 6241 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc 6301 agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg 6361 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt 6421 tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg tgttgccacc tggattctgc 6481 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg 6541 gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga 6601 tctccctttg ggccgcctcc ccgcctggaa ttaattctgc agtcgagacc tagaaaaaca 6661 tggagcaatc acaagtagca atacagcagc taccaatgct gattgtgcct ggctagaagc 6721 acaagaggag gaggaggtgg gttttccagt cacacctcag gtacctttaa gaccaatgac 6781 ttacaaggca gctgtagatc ttagccactt tttaaaagaa aagaggggac tggaagggct 6841 aattcactcc caacgaagac aagatatcct tgatctgtgg atctaccaca cacaaggcta 6901 cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac tgacctttgg 6961 atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca ataaaggaga 7021 gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg agagagaagt 7081 gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag agctgcatcc 7141 ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg ctggggactt 7201 tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat cctgcatata 7261 agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga gcctgggagc 7321 tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc 7381 aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt 7441 agtcagtgtg gaaaatctct agcagtagta gttcatgtca tcttattatt cagtatttat 7501 aacttgcaaa gaaatgaata tcagagagtg agaggccttg acattgctag cgtttaccgt 7561 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt 7621 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg 7681 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg 7741 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc 7801 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc 7861 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata 7921 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg 7981 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct 8041 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa 8101 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc 8161 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt 8221 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg 8281 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg 8341 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct 8401 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc 8461 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg 8521 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc 8581 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt 8641 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa 8701 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat 8761 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct 8821 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg 8881 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag 8941 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta 9001 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg 9061 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg 9121 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct 9181 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta 9241 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg 9301 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc 9361 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg 9421 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga 9481 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg 9541 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat 9601 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc 9661 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca 9721 catttccccg aaaagtgcca cctgacgtcg acggatcggg agatcaactt gtttattgca 9781 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt 9841 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc 9901 aactggataa ctcaagctaa ccaaaatcat cccaaacttc ccaccccata ccctattacc 9961 actgccaatt acctgtggtt tcatttactc taaacctgtg attcctctga attattttca 10021 ttttaaagaa attgtatttg ttaaatatgt actacaaact tagtagtttt taaagaaatt 10081 gtatttgtta aatatgtact acaaacttag tagt //