LOCUS V043068 9496 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V043068 VERSION V043068 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..3522 /label="Casp1(NM_009807)" /note="Casp1(NM_009807)" /gene="Casp1" CDS 3529..3594 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3638..3849 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3862..4130 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4159..4168 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4189..4851 /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 4921..4974 /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 4975..5574 /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 5575..6163 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6237..6470 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6542..6663 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6703..6838 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6871..6887) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6895..6911 /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(6919..6949) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6964..6985 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7273..7861) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8032..8892) /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(8893..8997) /gene="bla" /label="AmpR promoter" primer_bind 9471..9487 /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 ctgacaagat cctgagggca 2341 aagaggaagc aatttatcaa ctcagtgagt atagggacaa taaatggatt gttggatgaa 2401 cttttagaga agagagtgct gaatcaggaa gaaatggata aaataaaact tgcaaacatt 2461 actgctatgg acaaggcacg ggacctatgt gatcatgtct ctaaaaaagg gccccaggca 2521 agccaaatct ttatcactta catttgtaat gaagactgct acctggcagg aattctggag 2581 cttcaatcag ctccatcagc tgaaacattt gttgctacag aagattctaa aggaggacat 2641 ccttcatcct cagaaacaaa ggaagaacag aacaaagaag atggcacatt tccaggactg 2701 actgggaccc tcaagttttg ccctttagaa aaagcccaga agttatggaa agaaaatcct 2761 tcagagattt atccaataat gaatacaacc actcgtacac gtcttgccct cattatctgc 2821 aacacagagt ttcaacatct ttctccgagg gttggagctc aagttgacct cagagaaatg 2881 aagttgctgc tggaggatct ggggtatacc gtgaaagtga aagaaaatct cacagctctg 2941 gagatggtga aagaggtgaa agaatttgct gcctgcccag agcacaagac ttctgacagt 3001 actttccttg tattcatgtc tcatggtatc caggagggaa tatgtgggac cacatactct 3061 aatgaagttt cagatatttt aaaggttgac acaatctttc agatgatgaa cactttgaag 3121 tgcccaagct tgaaagacaa gcccaaggtg atcattattc aggcatgccg tggagagaaa 3181 caaggagtgg tgttgttaaa agattcagta agagactctg aagaggattt cttaacggat 3241 gcaatttttg aagatgatgg cattaagaag gcccatatag agaaagattt tattgctttc 3301 tgctcttcaa caccagataa tgtgtcttgg agacatcctg tcaggggctc acttttcatt 3361 gagtcactca tcaaacacat gaaagaatat gcctggtctt gtgacttgga ggacattttc 3421 agaaaggttc gattttcatt tgaacaacca gaatttaggc tacagatgcc cactgctgat 3481 agggtgaccc tgacaaaacg tttctacctc ttcccgggac atggatccga ctacaaagac 3541 catgacggtg attataaaga tcatgacatc gactacaagg atgacgatga caagtagcgg 3601 ccgcgaagga tctgcgatcg ctccggtgcc cgtcagtggg cagagcgcac atcgcccaca 3661 gtccccgaga agttgggggg aggggtcggc aattgaacgg gtgcctagag aaggtggcgc 3721 ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc tttttcccga gggtggggga 3781 gaaccgtata taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg gtttgccgcc 3841 agaacacagc tgaagcttcg aggggctcgc atctctcctt cacgcgcccg ccgccctacc 3901 tgaggccgcc atccacgccg gttgagtcgc gttctgccgc ctcccgcctg tggtgcctcc 3961 tgaactgcgt ccgccgtcta ggtaagttta aagctcaggt cgagaccggg cctttgtccg 4021 gcgctccctt ggagcctacc tagactcagc cggctctcca cgctttgcct gaccctgctt 4081 gctcaactct acgtctttgt ttcgttttct gttctgcgcc gttacagatc caagctgtga 4141 ccggcgccta cgctagacgc caccatggag agcgacgaga gcggcctgcc cgccatggag 4201 atcgagtgcc gcatcaccgg caccctgaac ggcgtggagt tcgagctggt gggcggcgga 4261 gagggcaccc ccaagcaggg ccgcatgacc aacaagatga agagcaccaa aggcgccctg 4321 accttcagcc cctacctgct gagccacgtg atgggctacg gcttctacca cttcggcacc 4381 taccccagcg gctacgagaa ccccttcctg cacgccatca acaacggcgg ctacaccaac 4441 acccgcatcg agaagtacga ggacggcggc gtgctgcacg tgagcttcag ctaccgctac 4501 gaggccggcc gcgtgatcgg cgacttcaag gtggtgggca ccggcttccc cgaggacagc 4561 gtgatcttca ccgacaagat catccgcagc aacgccaccg tggagcacct gcaccccatg 4621 ggcgataacg tgctggtggg cagcttcgcc cgcaccttca gcctgcgcga cggcggctac 4681 tacagcttcg tggtggacag ccacatgcac ttcaagagcg ccatccaccc cagcatcctg 4741 cagaacgggg gccccatgtt cgccttccgc cgcgtggagg agctgcacag caacaccgag 4801 ctgggcatcg tggagtacca gcacgccttc aagaccccca tcgccttcgc cagatcccgc 4861 gctcagtcgt ccaattctgc cgtggacggc accgccggac ccggctccac cggatctcgc 4921 gagggcagag gaagtcttct aacatgcggt gacgtggagg agaatcccgg ccctatgacc 4981 gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tccccagggc cgtacgcacc 5041 ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac 5101 atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc 5161 aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc 5221 gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg 5281 ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg 5341 tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc 5401 gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc 5461 tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc 5521 gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctgaaatcaa 5581 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt 5641 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct 5701 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc 5761 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg 5821 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc 5881 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc 5941 actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt 6001 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca 6061 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt 6121 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggtac ctttaagacc 6181 aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg ggggactgga 6241 agggctaatt cactcccaac gaagataaga tctgcttttt gcttgtactg ggtctctctg 6301 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc 6361 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg 6421 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gtagtagttc 6481 atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag agagtgagag 6541 gaacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac 6601 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc 6661 ttatcatgtc tggctctagc tatcccgccc ctaactccgc ccatcccgcc cctaactccg 6721 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc 6781 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta 6841 gacttttgca gagaccaaat tcgtaatcat gtcatagctg tttcctgtgt gaaattgtta 6901 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc 6961 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg 7021 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg 7081 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg 7141 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa 7201 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc 7261 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc 7321 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag 7381 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct 7441 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta 7501 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc 7561 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc 7621 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt 7681 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct 7741 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc 7801 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca 7861 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta 7921 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa 7981 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg 8041 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg 8101 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc 8161 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc 8221 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa 8281 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc 8341 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg 8401 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc 8461 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat 8521 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg 8581 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc 8641 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg 8701 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat 8761 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg 8821 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg 8881 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct 8941 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac 9001 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta 9061 taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa 9121 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag 9181 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta 9241 tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag 9301 atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg 9361 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc 9421 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac 9481 ggccagtgcc aagctg //