LOCUS V020921 9518 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V020921 VERSION V020921 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 2826..4214 /label="CTSC(NM_001814)" /note="CTSC(NM_001814)" /gene="CTSC" CDS 4221..4286 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 4306..4805 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 4826..5425 /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 5439..6027 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6234..6867 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(6996..7012) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7020..7036 /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(7044..7074) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7089..7110 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7398..7986) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8157..9017) /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(9018..9122) /gene="bla" /label="AmpR promoter" polyA_signal 9170..9304 /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 ccaccatggg tgctgggccc tccttgctgc tcgccgccct cctgctgctt ctctccggcg 2881 acggcgccgt gcgctgcgac acacctgcca actgcaccta tcttgacctg ctgggcacct 2941 gggtcttcca ggtgggctcc agcggttccc agcgcgatgt caactgctcg gttatgggac 3001 cacaagaaaa aaaagtagtg gtgtaccttc agaagctgga tacagcatat gatgaccttg 3061 gcaattctgg ccatttcacc atcatttaca accaaggctt tgagattgtg ttgaatgact 3121 acaagtggtt tgcctttttt aagtataaag aagagggcag caaggtgacc acttactgca 3181 acgagacaat gactgggtgg gtgcatgatg tgttgggccg gaactgggct tgtttcaccg 3241 gaaagaaggt gggaactgcc tctgagaatg tgtatgtcaa catagcacac cttaagaatt 3301 ctcaggaaaa gtattctaat aggctctaca agtatgatca caactttgtg aaagctatca 3361 atgccattca gaagtcttgg actgcaacta catacatgga atatgagact cttaccctgg 3421 gagatatgat taggagaagt ggtggccaca gtcgaaaaat cccaaggccc aaacctgcac 3481 cactgactgc tgaaatacag caaaagattt tgcatttgcc aacatcttgg gactggagaa 3541 atgttcatgg tatcaatttt gtcagtcctg ttcgaaacca agcatcctgt ggcagctgct 3601 actcatttgc ttctatgggt atgctagaag cgagaatccg tatactaacc aacaattctc 3661 agaccccaat cctaagccct caggaggttg tgtcttgtag ccagtatgct caaggctgtg 3721 aaggcggctt cccatacctt attgcaggaa agtacgccca agattttggg ctggtggaag 3781 aagcttgctt cccctacaca ggcactgatt ctccatgcaa aatgaaggaa gactgctttc 3841 gttattactc ctctgagtac cactatgtag gaggtttcta tggaggctgc aatgaagccc 3901 tgatgaagct tgagttggtc catcatgggc ccatggcagt tgcttttgaa gtatatgatg 3961 acttcctcca ctacaaaaag gggatctacc accacactgg tctaagagac cctttcaacc 4021 cctttgagct gactaatcat gctgttctgc ttgtgggcta tggcactgac tcagcctctg 4081 ggatggatta ctggattgtt aaaaacagct ggggcaccgg ctggggtgag aatggctact 4141 tccggatccg cagaggaact gatgagtgtg caattgagag catagcagtg gcagccacac 4201 caattcctaa attgggatcc gactacaaag accatgacgg tgattataaa gatcatgaca 4261 tcgactacaa ggatgacgat gacaagtaat ctagataatt ctaccgggta ggggaggcgc 4321 ttttcccaag gcagtctgga gcatgcgctt tagcagcccc gctgggcact tggcgctaca 4381 caagtggcct ctggcctcgc acacattcca catccaccgg taggcgccaa ccggctccgt 4441 tctttggtgg ccccttcgcg ccaccttcta ctcctcccct agtcaggaag ttcccccccg 4501 ccccgcagct cgcgtcgtgc aggacgtgac aaatggaagt agcacgtctc actagtctcg 4561 tgcagatgga cagcaccgct gagcaatgga agcgggtagg cctttggggc agcggccaat 4621 agcagctttg ctccttcgct ttctgggctc agaggctggg aaggggtggg tccgggggcg 4681 ggctcagggg cgggctcagg ggcggggcgg gcgcccgaag gtcctccgga ggcccggcat 4741 tctgcacgct tcaaaagcgc acgtctgccg cgctgttctc ctcttcctca tctccgggcc 4801 tttcgacctg cagcccaagc ttaccatgac cgagtacaag cccacggtgc gcctcgccac 4861 ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc 4921 cacgcgccac accgtcgatc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact 4981 cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc 5041 ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg 5101 cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct 5161 cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc 5221 cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga 5281 gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga 5341 gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg 5401 catgacccgc aagcccggtg cctgaccgcg tctggaacaa tcaacctctg gattacaaaa 5461 tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg 5521 ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct 5581 tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg 5641 gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct 5701 gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg 5761 ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg 5821 tgttgtcggg gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc acctggattc 5881 tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc 5941 gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc 6001 ggatctccct ttgggccgcc tccccgcctg gaattaattc tgcagtcgag acctagaaaa 6061 acatggagca atcacaagta gcaatacagc agctaccaat gctgattgtg cctggctaga 6121 agcacaagag gaggaggagg tgggttttcc agtcacacct caggtacctt taagaccaat 6181 gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaagaggg gactggaagg 6241 gctaattcac tcccaacgaa gacaagatat ccttgatctg tggatctacc acacacaagg 6301 ctacttccct gattagcaga actacacacc agggccaggg gtcagatatc cactgacctt 6361 tggatggtgc tacaagctag taccagttga gccagataag gtagaagagg ccaataaagg 6421 agagaacacc agcttgttac accctgtgag cctgcatggg atggatgacc cggagagaga 6481 agtgttagag tggaggtttg acagccgcct agcatttcat cacgtggccc gagagctgca 6541 tccggagtac ttcaagaact gctgatatcg agcttgctac aagggacttt ccgctgggga 6601 ctttccaggg aggcgtggcc tgggcgggac tggggagtgg cgagccctca gatcctgcat 6661 ataagcagct gctttttgcc tgtactgggt ctctctggtt agaccagatc tgagcctggg 6721 agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc 6781 ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct 6841 tttagtcagt gtggaaaatc tctagcagta gtagttcatg tcatcttatt attcagtatt 6901 tataacttgc aaagaaatga atatcagaga gtgagaggcc ttgacattgc tagcgtttta 6961 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat 7021 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg 7081 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag 7141 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt 7201 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg 7261 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg 7321 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag 7381 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga 7441 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct 7501 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc 7561 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg 7621 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc 7681 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca 7741 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag 7801 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct 7861 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc 7921 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga 7981 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca 8041 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat 8101 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac 8161 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt 8221 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt 8281 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag 8341 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct 8401 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt 8461 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc 8521 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt 8581 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg 8641 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg 8701 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct 8761 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc 8821 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt 8881 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt 8941 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg 9001 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat 9061 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg 9121 cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatca acttgtttat 9181 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt 9241 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg 9301 gatcaactgg ataactcaag ctaaccaaaa tcatcccaaa cttcccaccc cataccctat 9361 taccactgcc aattacctgt ggtttcattt actctaaacc tgtgattcct ctgaattatt 9421 ttcattttaa agaaattgta tttgttaaat atgtactaca aacttagtag tttttaaaga 9481 aattgtattt gttaaatatg tactacaaac ttagtagt //