LOCUS V034177 8984 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V034177 VERSION V034177 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 2323..2985 /label="BAX(NM_001291428)" /note="BAX(NM_001291428)" /gene="BAX" CDS 2992..3018 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" CDS 3022..3048 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" CDS 3052..3078 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" promoter 3143..3354 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3367..3635 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 3664..3673 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 3694..4356 /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 4426..4479 /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 4480..5079 /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 5080..5667 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5741..5974 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6046..6167 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6186..6321 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6359..6375) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6383..6399 /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(6407..6437) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6452..6473 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6761..7349) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7520..8380) /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(8381..8485) /gene="bla" /label="AmpR promoter" primer_bind 8959..8975 /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 ggatccgcca ccatggacgg gtccggggag 2341 cagcccagag gcggggggcc caccagctct gagcagatca tgaagacagg ggcccttttg 2401 cttcagggtt tcatccagga tcgagcaggg cgaatggggg gggaggcacc cgagctggcc 2461 ctggacccgg tgcctcagga tgcgtccacc aagaagctga gcgagtgtct caagcgcatc 2521 ggggacgaac tggacagtaa catggagctg cagaggatga ttgccgccgt ggacacagac 2581 tccccccgag aggtcttttt ccgagtggca gctgacatgt tttctgacgg caacttcaac 2641 tggggccggg ttgtcgccct tttctacttt gccagcaaac tggtgctcaa ggccctgtgc 2701 accaaggtgc cggaactgat cagaaccatc atgggctgga cattggactt cctccgggag 2761 cggctgttgg gctggatcca agaccagggt ggttgggggc tgcccctggc cgagtcactg 2821 aagcgactga tgtccctgtc tccaggacgg cctcctctcc tactttggga cgcccacgtg 2881 gcagaccgtg accatctttg tggcgggagt gctcaccgcc tcactcacca tctggaagaa 2941 gatgggctga ggcccccagc tgccttggac tgtgtttttc ctccactcga gtacccatac 3001 gacgtacctg attatgcaac ttatccttat gatgtacctg attatgctag ctatccttat 3061 gatgtcccag actacgcata gggatccctg cgatatccag cgcggccgcg aaggatctgc 3121 gatcgctccg gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc cgagaagttg 3181 gggggagggg tcggcaattg aacgggtgcc tagagaaggt ggcgcggggt aaactgggaa 3241 agtgatgtcg tgtactggct ccgccttttt cccgagggtg ggggagaacc gtatataagt 3301 gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg ccgccagaac acagctgaag 3361 cttcgagggg ctcgcatctc tccttcacgc gcccgccgcc ctacctgagg ccgccatcca 3421 cgccggttga gtcgcgttct gccgcctccc gcctgtggtg cctcctgaac tgcgtccgcc 3481 gtctaggtaa gtttaaagct caggtcgaga ccgggccttt gtccggcgct cccttggagc 3541 ctacctagac tcagccggct ctccacgctt tgcctgaccc tgcttgctca actctacgtc 3601 tttgtttcgt tttctgttct gcgccgttac agatccaagc tgtgaccggc gcctacgcta 3661 gacgccacca tggagagcga cgagagcggc ctgcccgcca tggagatcga gtgccgcatc 3721 accggcaccc tgaacggcgt ggagttcgag ctggtgggcg gcggagaggg cacccccaag 3781 cagggccgca tgaccaacaa gatgaagagc accaaaggcg ccctgacctt cagcccctac 3841 ctgctgagcc acgtgatggg ctacggcttc taccacttcg gcacctaccc cagcggctac 3901 gagaacccct tcctgcacgc catcaacaac ggcggctaca ccaacacccg catcgagaag 3961 tacgaggacg gcggcgtgct gcacgtgagc ttcagctacc gctacgaggc cggccgcgtg 4021 atcggcgact tcaaggtggt gggcaccggc ttccccgagg acagcgtgat cttcaccgac 4081 aagatcatcc gcagcaacgc caccgtggag cacctgcacc ccatgggcga taacgtgctg 4141 gtgggcagct tcgcccgcac cttcagcctg cgcgacggcg gctactacag cttcgtggtg 4201 gacagccaca tgcacttcaa gagcgccatc caccccagca tcctgcagaa cgggggcccc 4261 atgttcgcct tccgccgcgt ggaggagctg cacagcaaca ccgagctggg catcgtggag 4321 taccagcacg ccttcaagac ccccatcgcc ttcgccagat cccgcgctca gtcgtccaat 4381 tctgccgtgg acggcaccgc cggacccggc tccaccggat ctcgcgaggg cagaggaagt 4441 cttctaacat gcggtgacgt ggaggagaat cccggcccta tgaccgagta caagcccacg 4501 gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc 4561 gccgactacc ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc 4621 gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg 4681 gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg 4741 ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa 4801 cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc 4861 gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga 4921 gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac 4981 ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga 5041 ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgaa atcaacctct ggattacaaa 5101 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac 5161 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc 5221 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt 5281 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc 5341 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc 5401 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg 5461 gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt 5521 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc 5581 cgcggcctgc tgccggctct gcggcctctt ccgcgtctcc gccttcgccc tcagacgagt 5641 cggatctccc tttggccgcc tccccgcctg gtacctttaa gaccaatgac ttacaaggca 5701 gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc 5761 caacgaaaat aagatctgct ttttgcttgt actgggtctc tctggttaga ccagatctga 5821 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct 5881 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc 5941 agaccctttt agtcagtgtg gaaaatctct agcagtagta gttcatgtca tcttattatt 6001 cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggaactt gtttattgca 6061 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt 6121 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggctc 6181 tagctatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa 6241 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt 6301 gaggaggctt ttttggaggc ctagactttt gcagagacgg cccaaattcg taatcatggt 6361 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg 6421 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt 6481 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg 6541 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg 6601 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa 6661 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc 6721 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc 6781 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat 6841 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc 6901 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct 6961 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg 7021 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc 7081 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga 7141 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa 7201 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta 7261 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc 7321 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg 7381 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga 7441 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg 7501 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct 7561 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg 7621 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc 7681 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa 7741 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc 7801 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt 7861 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc 7921 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt 7981 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc 8041 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt 8101 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata 8161 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga 8221 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag 8281 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa 8341 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt 8401 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga 8461 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag 8521 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc 8581 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca 8641 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg 8701 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc 8761 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc 8821 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat 8881 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt 8941 tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa gctg //