LOCUS V018335 11206 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V018335 VERSION V018335 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 2827..5925 /label="Nlrp3(NM_145827)" /note="Nlrp3(NM_145827)" /gene="Nlrp3" CDS 5926..5955 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /transl_table=1 /translation="EQKLISEEDL" /label="Myc" promoter 5994..6493 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 6514..7113 /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 7127..7715 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 7922..8555 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(8684..8700) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 8708..8724 /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(8732..8762) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 8777..8798 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(9086..9674) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(9845..10705) /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(10706..10810) /gene="bla" /label="AmpR promoter" polyA_signal 10858..10992 /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 gccaccatga cgagtgtccg ttgcaagctg gctcagtatc tagaggacct tgaagatgtg 2881 gacctcaaga aattcaaaat gcatttggaa gattacccgc ccgagaaagg ctgtatccca 2941 gtccccaggg gccagatgga gaaggcagat cacttggatc tagccacact catgattgac 3001 ttcaatggcg aggagaaggc ctgggccatg gctgtgtgga tctttgctgc gatcaacagg 3061 cgagacctct gggaaaaagc taagaaggac cagccagagt ggaatgacac gtgtacatca 3121 cattcctcta tggtatgcca ggaggacagc cttgaagaag agtggatggg tttgctggga 3181 tatctctccc gcatctccat ttgtaaaaag aagaaagatt actgtaagat gtacagacga 3241 catgtgagaa gcaggttcta ctctatcaag gacaggaacg cgcgtctagg tgagagtgtg 3301 gacctcaaca gtcgctacac gcagctccaa ctggtcaagg agcatccaag caagcaggag 3361 cgggagcatg aactcctgac catcggccgg actaaaatgc gggacagccc catgagttcc 3421 cttaagctgg agctgctgtt tgagcccgag gacgggcact cggagcctgt gcacacagtg 3481 gtgttccagg gagcagcagg catcgggaaa accatcctag ccaggaagat tatgttggac 3541 tgggcactgg gaaagctctt caaagacaaa tttgactatt tgttctttat ccactgccga 3601 gaggtgagcc tcaggacgcc aaggagtcta gcagacctga ttgtcagctg ctggcctgac 3661 ccaaacccac cagtgtgcaa gatcctgcgc aagccttcca ggatcctctt cctcatggat 3721 ggctttgatg agctacaagg ggcctttgac gagcacattg gggaggtctg cacagactgg 3781 caaaaggctg tgcggggaga cattctgcta agcagcctca tccgaaagaa actgctgccc 3841 aaggcctctc tgctcataac gacgaggccg gtagccttgg agaaactgca gcatctcctg 3901 gaccaccccc gccatgtgga gatcctaggt ttctctgagg ccaaaaggaa ggagtatttc 3961 tttaagtatt tctccaacga gctgcaggcc cgggaggcct tcaggctgat ccaagagaat 4021 gaggtcctct ttaccatgtg cttcatcccc ctggtctgct ggattgtgtg cacggggcta 4081 aagcaacaga tggagaccgg gaagagcctg gcccagacct ccaagaccac tacggccgtc 4141 tacgtcttct tcctttccag cctgctgcaa tcccgggggg gcattgagga gcatctcttc 4201 tctgactacc tacaggggct ctgttcactg gctgcggatg gaatttggaa ccagaaaatc 4261 ctatttgagg agtgtgatct gcggaagcac ggcctgcaga agactgacgt ctccgctttc 4321 ctgaggatga acgtgttcca gaaggaagtg gactgcgaga gattctacag cttcagccac 4381 atgactttcc aggagttctt cgctgctatg tactatttgc tggaagagga ggcagagggg 4441 gagaccgtga ggaaaggacc aggaggttgt tcagatcttc tgaaccgaga cgtgaaggtc 4501 ctactagaaa attacggcaa gtttgaaaaa ggctatctga tttttgttgt ccgattcctc 4561 tttggccttg taaaccagga gagaacctct tatttggaga agaaactaag ttgcaagatc 4621 tctcagcaag tcagactgga actactgaag tggattgaag tgaaagccaa ggccaagaag 4681 ctgcagtggc agcccagcca actggaactg ttctactgcc tgtacgagat gcaggaggaa 4741 gactttgtgc agagtgccat ggaccacttt cccaaaattg agatcaacct ctctaccaga 4801 atggaccacg tggtttcctc cttttgtatt aagaactgtc atagggtcaa aacgctttcc 4861 ctgggttttt ttcacaactc gcccaaggag gaagaagaag agaggagagg aggtcgaccc 4921 ttggaccagg ttcagtgtgt tttcccagac actcatgttg cctgttcttc cagactggtg 4981 aactgctgcc tcacttctag cttctgccgt ggtctcttct caagtctaag caccaaccgg 5041 agcctcactg aactggacct cagtgacaat actctgggag acccgggcat gagggtgctg 5101 tgtgaggcac tccagcaccc aggctgtaac attcagagac tgtggttggg gcgctgcgga 5161 ctgtcccatc aatgctgctt cgacatctcc tctgtcctga gcagcagcca gaagctggtg 5221 gagctggacc tcagtgacaa tgccctgggg gactttggaa tcagattgct gtgtgtggga 5281 ctgaagcacc tgctctgcaa cctccagaaa ctgtggttgg tgagctgctg tctcacatcc 5341 gcgtgttgtc aggatctcgc attggttctg agctccaacc attctctgac cagactgtac 5401 attggagaaa atgccttggg agactcagga gtccaagttt tgtgtgaaaa gatgaaggac 5461 ccacagtgta acttgcagaa gctggggttg gtgaattccg gccttacttc aatctgttgt 5521 tcagctctga cctctgtgct caaaaccaac cagaacttca cacacctcta tctacgaagc 5581 aatgcccttg gagacacagg actcaggctc ctctgtgagg ggcttctgca cccggactgt 5641 aaactacaga tgctggaatt agacaactgc agcctcacct cacacagctg ctggaatctc 5701 tccacaattc tgacccacaa ccacagcctt cggaagctga acctgggcaa caatgatctt 5761 ggcgatctgt gcgtggtgac cctctgtgag gtgctgaaac agcagggctg cctcctgcag 5821 agcctacagt tgggtgaaat gtacttaaat cgtgaaacaa aacgtgcctt agaagcgctc 5881 caggaagaaa agcctgagct gactatagtc ttcgagattt cctgggaaca aaaactcatc 5941 tcagaagagg atctgtgagg gcccgggatc ccgcgactct agataattct accgggtagg 6001 ggaggcgctt ttcccaaggc agtctggagc atgcgcttta gcagccccgc tgggcacttg 6061 gcgctacaca agtggcctct ggcctcgcac acattccaca tccaccggta ggcgccaacc 6121 ggctccgttc tttggtggcc ccttcgcgcc accttctact cctcccctag tcaggaagtt 6181 cccccccgcc ccgcagctcg cgtcgtgcag gacgtgacaa atggaagtag cacgtctcac 6241 tagtctcgtg cagatggaca gcaccgctga gcaatggaag cgggtaggcc tttggggcag 6301 cggccaatag cagctttgct ccttcgcttt ctgggctcag aggctgggaa ggggtgggtc 6361 cgggggcggg ctcaggggcg ggctcagggg cggggcgggc gcccgaaggt cctccggagg 6421 cccggcattc tgcacgcttc aaaagcgcac gtctgccgcg ctgttctcct cttcctcatc 6481 tccgggcctt tcgacctgca gcccaagctt accatgaccg agtacaagcc cacggtgcgc 6541 ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac 6601 taccccgcca cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt caccgagctg 6661 caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt cgcggacgac 6721 ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc 6781 gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca gcaacagatg 6841 gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc caccgtcggc 6901 gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc cggagtggag 6961 gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg caacctcccc 7021 ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc 7081 acctggtgca tgacccgcaa gcccggtgcc tgaccgcgtc tggaacaatc aacctctgga 7141 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg 7201 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt 7261 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag 7321 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc 7381 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga 7441 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa 7501 ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac 7561 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct 7621 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca 7681 gacgagtcgg atctcccttt gggccgcctc cccgcctgga attaattctg cagtcgagac 7741 ctagaaaaac atggagcaat cacaagtagc aatacagcag ctaccaatgc tgattgtgcc 7801 tggctagaag cacaagagga ggaggaggtg ggttttccag tcacacctca ggtaccttta 7861 agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga aaagagggga 7921 ctggaagggc taattcactc ccaacgaaga caagatatcc ttgatctgtg gatctaccac 7981 acacaaggct acttccctga ttagcagaac tacacaccag ggccaggggt cagatatcca 8041 ctgacctttg gatggtgcta caagctagta ccagttgagc cagataaggt agaagaggcc 8101 aataaaggag agaacaccag cttgttacac cctgtgagcc tgcatgggat ggatgacccg 8161 gagagagaag tgttagagtg gaggtttgac agccgcctag catttcatca cgtggcccga 8221 gagctgcatc cggagtactt caagaactgc tgatatcgag cttgctacaa gggactttcc 8281 gctggggact ttccagggag gcgtggcctg ggcgggactg gggagtggcg agccctcaga 8341 tcctgcatat aagcagctgc tttttgcctg tactgggtct ctctggttag accagatctg 8401 agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc 8461 ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct 8521 cagacccttt tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc atcttattat 8581 tcagtattta taacttgcaa agaaatgaat atcagagagt gagaggcctt gacattgcta 8641 gcgttttacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg 8701 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta 8761 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg 8821 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga 8881 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg 8941 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag 9001 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc 9061 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca 9121 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt 9181 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc 9241 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc 9301 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc 9361 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact 9421 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg 9481 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta 9541 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca 9601 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa 9661 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg 9721 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc 9781 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg 9841 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat 9901 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg 9961 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa 10021 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca 10081 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc 10141 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt 10201 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa 10261 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat 10321 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct 10381 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga 10441 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag 10501 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga 10561 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca 10621 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg 10681 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc 10741 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag 10801 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatcaac 10861 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat 10921 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat 10981 catgtctgga tcaactggat aactcaagct aaccaaaatc atcccaaact tcccacccca 11041 taccctatta ccactgccaa ttacctgtgg tttcatttac tctaaacctg tgattcctct 11101 gaattatttt cattttaaag aaattgtatt tgttaaatat gtactacaaa cttagtagtt 11161 tttaaagaaa ttgtatttgt taaatatgta ctacaaactt agtagt //