LOCUS V029828 8767 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V029828 VERSION V029828 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..2793 /label="RPL29(NM_000992)" /note="RPL29(NM_000992)" /gene="RPL29" CDS 2800..2865 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 2909..3120 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3133..3401 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 3430..3439 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 3460..4122 /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 4192..4245 /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 4246..4845 /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 4846..5434 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5508..5741 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 5813..5934 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 5974..6109 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6142..6158) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6166..6182 /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(6190..6220) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6235..6256 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6544..7132) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7303..8163) /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(8164..8268) /gene="bla" /label="AmpR promoter" primer_bind 8742..8758 /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 ccaagtccaa gaaccacacc 2341 acacacaacc agtcccgaaa atggcacaga aatggtatca agaaaccccg atcacaaaga 2401 tacgaatctc ttaagggggt ggaccccaag ttcctgagga acatgcgctt tgccaagaag 2461 cacaacaaaa agggcctaaa gaagatgcag gccaacaatg ccaaggccat gagtgcacgt 2521 gccgaggcta tcaaggccct cgtaaagccc aaggaggtta agcccaagat cccaaagggt 2581 gtcagccgca agctcgatcg acttgcctac attgcccacc ccaagcttgg gaagcgtgct 2641 cgtgcccgta ttgccaaggg gctcaggctg tgccggccaa aggccaaggc caaggccaag 2701 gccaaggatc aaaccaaggc ccaggctgca gccccagctt cagttccagc tcaggctccc 2761 aaacgtaccc aggcccctac aaaggcttca gagggatccg actacaaaga ccatgacggt 2821 gattataaag atcatgacat cgactacaag gatgacgatg acaagtagcg gccgcgaagg 2881 atctgcgatc gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag 2941 aagttggggg gaggggtcgg caattgaacg ggtgcctaga gaaggtggcg cggggtaaac 3001 tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat 3061 ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag 3121 ctgaagcttc gaggggctcg catctctcct tcacgcgccc gccgccctac ctgaggccgc 3181 catccacgcc ggttgagtcg cgttctgccg cctcccgcct gtggtgcctc ctgaactgcg 3241 tccgccgtct aggtaagttt aaagctcagg tcgagaccgg gcctttgtcc ggcgctccct 3301 tggagcctac ctagactcag ccggctctcc acgctttgcc tgaccctgct tgctcaactc 3361 tacgtctttg tttcgttttc tgttctgcgc cgttacagat ccaagctgtg accggcgcct 3421 acgctagacg ccaccatgga gagcgacgag agcggcctgc ccgccatgga gatcgagtgc 3481 cgcatcaccg gcaccctgaa cggcgtggag ttcgagctgg tgggcggcgg agagggcacc 3541 cccaagcagg gccgcatgac caacaagatg aagagcacca aaggcgccct gaccttcagc 3601 ccctacctgc tgagccacgt gatgggctac ggcttctacc acttcggcac ctaccccagc 3661 ggctacgaga accccttcct gcacgccatc aacaacggcg gctacaccaa cacccgcatc 3721 gagaagtacg aggacggcgg cgtgctgcac gtgagcttca gctaccgcta cgaggccggc 3781 cgcgtgatcg gcgacttcaa ggtggtgggc accggcttcc ccgaggacag cgtgatcttc 3841 accgacaaga tcatccgcag caacgccacc gtggagcacc tgcaccccat gggcgataac 3901 gtgctggtgg gcagcttcgc ccgcaccttc agcctgcgcg acggcggcta ctacagcttc 3961 gtggtggaca gccacatgca cttcaagagc gccatccacc ccagcatcct gcagaacggg 4021 ggccccatgt tcgccttccg ccgcgtggag gagctgcaca gcaacaccga gctgggcatc 4081 gtggagtacc agcacgcctt caagaccccc atcgccttcg ccagatcccg cgctcagtcg 4141 tccaattctg ccgtggacgg caccgccgga cccggctcca ccggatctcg cgagggcaga 4201 ggaagtcttc taacatgcgg tgacgtggag gagaatcccg gccctatgac cgagtacaag 4261 cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc 4321 gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg 4381 gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg 4441 gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg 4501 gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg 4561 cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg 4621 gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc 4681 cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc 4741 cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc 4801 gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgaaatca acctctggat 4861 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt 4921 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc 4981 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg 5041 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc 5101 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa 5161 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat 5221 tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc 5281 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt 5341 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag 5401 acgagtcgga tctccctttg ggccgcctcc ccgcctggta cctttaagac caatgactta 5461 caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat 5521 tcactcccaa cgaagataag atctgctttt tgcttgtact gggtctctct ggttagacca 5581 gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag 5641 cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag 5701 atccctcaga cccttttagt cagtgtggaa aatctctagc agtagtagtt catgtcatct 5761 tattattcag tatttataac ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt 5821 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc 5881 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt 5941 ctggctctag ctatcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc 6001 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc 6061 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct agacttttgc 6121 agagaccaaa ttcgtaatca tgtcatagct gtttcctgtg tgaaattgtt atccgctcac 6181 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt 6241 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc 6301 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg 6361 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt 6421 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa 6481 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc 6541 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag 6601 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt 6661 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg 6721 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg 6781 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg 6841 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac 6901 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg 6961 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt 7021 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg 7081 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc 7141 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt 7201 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt 7261 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag 7321 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt 7381 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc 7441 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc 7501 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg 7561 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac 7621 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg 7681 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc 7741 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact 7801 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc 7861 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat 7921 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc 7981 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac 8041 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa 8101 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact 8161 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg 8221 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg 8281 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag 8341 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca 8401 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc 8461 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc 8521 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag 8581 gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg 8641 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg 8701 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtgc 8761 caagctg //