LOCUS V023713 9605 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V023713 VERSION V023713 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..3648 /label="Sirt1(NM_001372090)" /note="Sirt1(NM_001372090)" /gene="Sirt1" CDS 3655..3720 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3764..3975 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3988..4256 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4285..4294 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4315..4977 /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 5047..5100 /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 5101..5700 /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 5701..6288 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6362..6595 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6667..6788 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6807..6942 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6980..6996) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7004..7020 /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(7028..7058) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7073..7094 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7382..7970) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8141..9001) /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(9002..9106) /gene="bla" /label="AmpR promoter" primer_bind 9580..9596 /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 gccaccatgt gcctgtgcag tggaaggaaa 2341 gcaattttgg aaatatatcc cggacagttc cagccatctc tgtgtcacaa attcatagct 2401 ttgtcagata aggaaggaaa actacttcga aattatactc aaaatataga taccttggag 2461 caggttgcag gaatccaaag gatcattcag tgtcatggtt cctttgcaac agcatcttgc 2521 ctgatttgta aatacaaagt tgattgtgaa gctgttcgtg gagatatttt taatcaggta 2581 gttcctcggt gtcctaggtg cccagctgat gagccacttg ccatcatgaa gccagagatt 2641 gtcttctttg gtgaaaactt accagaacag tttcatagag ccatgaagta tgacaaagat 2701 gaagttgacc tcctcattgt tattgggtct tctctgaaag taagaccagt agcactaatt 2761 ccaagttcta taccccatga agtgcctcaa atattaataa atagggaacc tctgcctcat 2821 ctacattttg atgtagagct tcttggagac tgcgatgtca taattaatga gttgtgtcat 2881 aggttaggtg gcgagtatgc caaactttgt tgtaaccctg taaagctttc agaaattact 2941 gaaaaacctc cacgaacaca aaaggaattg gttcatttat cagagttgcc accaacacct 3001 cttcatattt cagaagactc aagttcacct gaaagaactg taccacaaga ctcttctgtg 3061 attgctacac ttgtagacca aacaataaag aacaaagttg acgatttaga agtatctgaa 3121 ccaaaaagtt gtgtggaaga aaaatcacag gaagtacaga cttataggaa tgttgagagc 3181 attaatgtgg aaaacccaga tttcaaggct gttggttcca gtactggaga caaaaatgaa 3241 agaacttctg ttgcagaaac agtgagaaaa tgctggccta atagacttgc aaaggagcag 3301 attagtaagc gtcttgacgg taatcaatac ttgtttgtac caccaaatcg ttacatattt 3361 catggtgctg aggtatactc agactctgaa gatgacgcct tatcctctag ttcctgtggc 3421 agtaacagtg acagtggcac atgccagagt ccaagtttag aagaaccctt ggaagatgaa 3481 agtgaaattg aagagttcta caatggcttg gaagatgatg ctgacagacc ggagtgtgct 3541 ggaggatctg gagctgacgg aggggatcaa gaggcagtta atgaagctat agccatgaaa 3601 caggaattaa cagatgtaaa ctgtacacca gacaaatcag agcactatgg atccgactac 3661 aaagaccatg acggtgatta taaagatcat gacatcgact acaaggatga cgatgacaag 3721 tagcggccgc gaaggatctg cgatcgctcc ggtgcccgtc agtgggcaga gcgcacatcg 3781 cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt gaacgggtgc ctagagaagg 3841 tggcgcgggg taaactggga aagtgatgtc gtgtactggc tccgcctttt tcccgagggt 3901 gggggagaac cgtatataag tgcagtagtc gccgtgaacg ttctttttcg caacgggttt 3961 gccgccagaa cacagctgaa gcttcgaggg gctcgcatct ctccttcacg cgcccgccgc 4021 cctacctgag gccgccatcc acgccggttg agtcgcgttc tgccgcctcc cgcctgtggt 4081 gcctcctgaa ctgcgtccgc cgtctaggta agtttaaagc tcaggtcgag accgggcctt 4141 tgtccggcgc tcccttggag cctacctaga ctcagccggc tctccacgct ttgcctgacc 4201 ctgcttgctc aactctacgt ctttgtttcg ttttctgttc tgcgccgtta cagatccaag 4261 ctgtgaccgg cgcctacgct agacgccacc atggagagcg acgagagcgg cctgcccgcc 4321 atggagatcg agtgccgcat caccggcacc ctgaacggcg tggagttcga gctggtgggc 4381 ggcggagagg gcacccccaa gcagggccgc atgaccaaca agatgaagag caccaaaggc 4441 gccctgacct tcagccccta cctgctgagc cacgtgatgg gctacggctt ctaccacttc 4501 ggcacctacc ccagcggcta cgagaacccc ttcctgcacg ccatcaacaa cggcggctac 4561 accaacaccc gcatcgagaa gtacgaggac ggcggcgtgc tgcacgtgag cttcagctac 4621 cgctacgagg ccggccgcgt gatcggcgac ttcaaggtgg tgggcaccgg cttccccgag 4681 gacagcgtga tcttcaccga caagatcatc cgcagcaacg ccaccgtgga gcacctgcac 4741 cccatgggcg ataacgtgct ggtgggcagc ttcgcccgca ccttcagcct gcgcgacggc 4801 ggctactaca gcttcgtggt ggacagccac atgcacttca agagcgccat ccaccccagc 4861 atcctgcaga acgggggccc catgttcgcc ttccgccgcg tggaggagct gcacagcaac 4921 accgagctgg gcatcgtgga gtaccagcac gccttcaaga cccccatcgc cttcgccaga 4981 tcccgcgctc agtcgtccaa ttctgccgtg gacggcaccg ccggacccgg ctccaccgga 5041 tctcgcgagg gcagaggaag tcttctaaca tgcggtgacg tggaggagaa tcccggccct 5101 atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc cagggccgta 5161 cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgatccggac 5221 cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac 5281 atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag 5341 agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt 5401 tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag 5461 cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc 5521 agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg 5581 gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc 5641 gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctga 5701 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct 5761 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt 5821 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg 5881 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact 5941 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct 6001 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg 6061 ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc atggctgctc 6121 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc 6181 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctc 6241 cgccttcgcc ctcagacgag tcggatctcc ctttggccgc ctccccgcct ggtaccttta 6301 agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga 6361 ctggaagggc taattcactc ccaacgaaaa taagatctgc tttttgcttg tactgggtct 6421 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt 6481 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac 6541 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtagt 6601 agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat atcagagagt 6661 gagaggaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat 6721 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat 6781 gtatcttatc atgtctggct ctagctatcc cgcccctaac tccgcccagt tccgcccatt 6841 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct 6901 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctagacttt tgcagagacg 6961 gcccaaattc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa 7021 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga 7081 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt 7141 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct 7201 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat 7261 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga 7321 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt 7381 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt 7441 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc 7501 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa 7561 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct 7621 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta 7681 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg 7741 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc 7801 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta 7861 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg 7921 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt 7981 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg 8041 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta 8101 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg 8161 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg 8221 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc 8281 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg 8341 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg 8401 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag 8461 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat 8521 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc 8581 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc 8641 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa 8701 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac 8761 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt 8821 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc 8881 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa 8941 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca 9001 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat 9061 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa 9121 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc 9181 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca 9241 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc 9301 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag 9361 agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga 9421 gaaaataccg catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat 9481 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat 9541 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgcca 9601 agctg //