LOCUS V025013 9725 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V025013 VERSION V025013 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers LTR 234..414 /note="truncated 5' long terminal repeat (LTR) from HIV-1" /label="5' LTR (truncated)" misc_feature 458..583 /note="packaging signal of human immunodeficiency virus type 1" /label="HIV-1 Ψ" misc_feature 1076..1309 /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." /label="RRE" CDS 1493..1537 /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 1804..1921 /note="central polypurine tract and central termination sequence of HIV-1" /label="cPPT/CTS" promoter 2011..2214 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 2317..3768 /label="GSDMD(NM_001166237)" /note="GSDMD(NM_001166237)" /gene="GSDMD" CDS 3775..3840 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3884..4095 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4108..4376 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4405..4414 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4435..5097 /codon_start=1 /product="green fluorescent protein 2 from Pontellina plumata, also known as ppluGFP2 (Shagin et al., 2004)" /transl_table=1 /translation="PAMEIECRITGTLNGVEFELVGGGEGTPKQGRMTNKMKSTKGALT FSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVLHVSFSYRY EAGRVIGDFKVVGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNVLVGSFARTFSLRDGG YYSFVVDSHMHFKSAIHPSILQNGGPMFAFRRVEELHSNTELGIVEYQHAFKTPIAFA" /label="CopGFP" CDS 5167..5220 /codon_start=1 /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /product="2A peptide from Thosea asigna virus capsid protein" /protein_id="" /transl_table=1 /translation="EGRGSLLTCGDVEENPGP" /label="T2A" CDS 5221..5820 /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 5821..6408 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6482..6715 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6787..6908 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6927..7062 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(7100..7116) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7124..7140 /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(7148..7178) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7193..7214 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7502..8090) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8261..9121) /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(9122..9226) /gene="bla" /label="AmpR promoter" primer_bind 9700..9716 /note="common sequencing primer, one of multiple similar variants" /label="M13 fwd" ORIGIN 1 acgcgtgtag tcttatgcaa tactcttgta gtcttgcaac atggtaacga tgagttagca 61 acatgcctta caaggagaga aaaagcaccg tgcatgccga ttggtggaag taaggtggta 121 cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca tggattggac gaaccactga 181 attgccgcat tgcagagata ttgtatttaa gtgcctagct cgatacaata aacgggtctc 241 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta 301 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact 361 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg 421 cccgaacagg gacctgaaag cgaaagggaa accagagctc tctcgacgca ggactcggct 481 tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt 541 gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag 601 aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt 661 aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt 721 agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg 781 atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag 841 gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag 901 taagaccacc gcacagcaag cggccactga tcttcagacc tggaggagga gatatgaggg 961 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag 1021 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag 1081 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcc tcaatgacgc 1141 tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga 1201 gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc 1261 aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg 1321 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata 1381 aatctctgga acagattgga atcacacgac ctggatggag tgggacagag aaattaacaa 1441 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga 1501 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa 1561 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat 1621 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt 1681 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg 1741 tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac ggtatcggtt 1801 aacttttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat 1861 aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt 1921 tatcgatact agtattatgc ccagtacatg accttatggg actttcctac ttggcagtac 1981 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg 2041 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg 2101 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca 2161 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggttt atataagcag agctcgttta 2221 gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagattc 2281 tagagctagc ctgatacgaa ctcggaattc gccaccatgg ggtcggcctt tgagcgggta 2341 gtccggagag tggtccagga gctggaccat ggtggggagt tcatccctgt gaccagcctg 2401 cagagctcca ctggcttcca gccctactgc ctggtggtta ggaagccctc aagctcatgg 2461 ttctggaaac cccgttataa gtgtgtcaac ctgtctatca aggacatcct ggagccggat 2521 gccgcggaac cagacgtgca gcgtggcagg agcttccact tctacgatgc catggatggg 2581 cagatacagg gcagcgtgga gctggcagcc ccaggacagg caaagatcgc aggcggggcc 2641 gcggtgtctg acagctccag cacctcaatg aatgtgtact cgctgagtgt ggaccctaac 2701 acctggcaga ctctgctcca tgagaggcac ctgcggcagc cagaacacaa agtcctgcag 2761 cagctgcgca gccgcgggga caacgtgtac gtggtgactg aggtgctgca gacacagaag 2821 gaggtggaag tcacgcgcac ccacaagcgg gagggctcgg gccggttttc cctgcccgga 2881 gccacgtgct tgcagggtga gggccagggc catctgagcc agaagaagac ggtcaccatc 2941 ccctcaggca gcaccctcgc attccgggtg gcccagctgg ttattgactc tgacttggac 3001 gtccttctct tcccggataa gaagcagagg accttccagc cacccgcgac aggccacaag 3061 cgttccacga gcgaaggcgc ctggccacag ctgccctctg gcctctccat gatgaggtgc 3121 ctccacaact tcctgacaga tggggtccct gcggaggggg cgttcactga agacttccag 3181 ggcctacggg cagaggtgga gaccatctcc aaggaactgg agcttttgga cagagagctg 3241 tgccagctgc tgctggaggg cctggagggg gtgctgcggg accagctggc cctgcgagcc 3301 ttggaggagg cgctggagca gggccagagc cttgggccgg tggagcccct ggacggtcca 3361 gcaggtgctg tcctggagtg cctggtgttg tcctccggaa tgctggtgcc ggaactcgct 3421 atccctgttg tctacctgct gggggcactg accatgctga gtgaaacgca gcacaagctg 3481 ctggcggagg cgctggagtc gcagaccctg ttggggccgc tcgagctggt gggcagcctc 3541 ttggagcaga gtgccccgtg gcaggagcgc agcaccatgt ccctgccccc cgggctcctg 3601 gggaacagct ggggcgaagg agcaccggcc tgggtcttgc tggacgagtg tggcctagag 3661 ctgggggagg acactcccca cgtgtgctgg gagccgcagg cccagggccg catgtgtgca 3721 ctctacgcct ccctggcact gctatcagga ctgagccagg agccccacgg atccgactac 3781 aaagaccatg acggtgatta taaagatcat gacatcgact acaaggatga cgatgacaag 3841 tagcggccgc gaaggatctg cgatcgctcc ggtgcccgtc agtgggcaga gcgcacatcg 3901 cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt gaacgggtgc ctagagaagg 3961 tggcgcgggg taaactggga aagtgatgtc gtgtactggc tccgcctttt tcccgagggt 4021 gggggagaac cgtatataag tgcagtagtc gccgtgaacg ttctttttcg caacgggttt 4081 gccgccagaa cacagctgaa gcttcgaggg gctcgcatct ctccttcacg cgcccgccgc 4141 cctacctgag gccgccatcc acgccggttg agtcgcgttc tgccgcctcc cgcctgtggt 4201 gcctcctgaa ctgcgtccgc cgtctaggta agtttaaagc tcaggtcgag accgggcctt 4261 tgtccggcgc tcccttggag cctacctaga ctcagccggc tctccacgct ttgcctgacc 4321 ctgcttgctc aactctacgt ctttgtttcg ttttctgttc tgcgccgtta cagatccaag 4381 ctgtgaccgg cgcctacgct agacgccacc atggagagcg acgagagcgg cctgcccgcc 4441 atggagatcg agtgccgcat caccggcacc ctgaacggcg tggagttcga gctggtgggc 4501 ggcggagagg gcacccccaa gcagggccgc atgaccaaca agatgaagag caccaaaggc 4561 gccctgacct tcagccccta cctgctgagc cacgtgatgg gctacggctt ctaccacttc 4621 ggcacctacc ccagcggcta cgagaacccc ttcctgcacg ccatcaacaa cggcggctac 4681 accaacaccc gcatcgagaa gtacgaggac ggcggcgtgc tgcacgtgag cttcagctac 4741 cgctacgagg ccggccgcgt gatcggcgac ttcaaggtgg tgggcaccgg cttccccgag 4801 gacagcgtga tcttcaccga caagatcatc cgcagcaacg ccaccgtgga gcacctgcac 4861 cccatgggcg ataacgtgct ggtgggcagc ttcgcccgca ccttcagcct gcgcgacggc 4921 ggctactaca gcttcgtggt ggacagccac atgcacttca agagcgccat ccaccccagc 4981 atcctgcaga acgggggccc catgttcgcc ttccgccgcg tggaggagct gcacagcaac 5041 accgagctgg gcatcgtgga gtaccagcac gccttcaaga cccccatcgc cttcgccaga 5101 tcccgcgctc agtcgtccaa ttctgccgtg gacggcaccg ccggacccgg ctccaccgga 5161 tctcgcgagg gcagaggaag tcttctaaca tgcggtgacg tggaggagaa tcccggccct 5221 atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc cagggccgta 5281 cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgatccggac 5341 cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac 5401 atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag 5461 agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt 5521 tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag 5581 cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc 5641 agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg 5701 gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc 5761 gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctga 5821 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct 5881 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt 5941 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg 6001 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact 6061 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct 6121 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg 6181 ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc atggctgctc 6241 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc 6301 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctc 6361 cgccttcgcc ctcagacgag tcggatctcc ctttggccgc ctccccgcct ggtaccttta 6421 agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga 6481 ctggaagggc taattcactc ccaacgaaaa taagatctgc tttttgcttg tactgggtct 6541 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt 6601 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac 6661 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtagt 6721 agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat atcagagagt 6781 gagaggaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat 6841 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat 6901 gtatcttatc atgtctggct ctagctatcc cgcccctaac tccgcccagt tccgcccatt 6961 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct 7021 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctagacttt tgcagagacg 7081 gcccaaattc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa 7141 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga 7201 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt 7261 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct 7321 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat 7381 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga 7441 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt 7501 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt 7561 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc 7621 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa 7681 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct 7741 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta 7801 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg 7861 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc 7921 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta 7981 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg 8041 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt 8101 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg 8161 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta 8221 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg 8281 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg 8341 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc 8401 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg 8461 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg 8521 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag 8581 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat 8641 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc 8701 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc 8761 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa 8821 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac 8881 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt 8941 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc 9001 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa 9061 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca 9121 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat 9181 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa 9241 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc 9301 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca 9361 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc 9421 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag 9481 agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga 9541 gaaaataccg catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat 9601 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat 9661 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgcca 9721 agctg //