LOCUS V038979 9307 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V038979 VERSION V038979 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..3333 /label="LSP1(NM_002339)" /note="LSP1(NM_002339)" /gene="LSP1" CDS 3340..3405 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3449..3660 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3673..3941 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 3970..3979 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4000..4662 /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 4732..4785 /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 4786..5385 /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 5386..5974 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6048..6281 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6353..6474 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6514..6649 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6682..6698) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6706..6722 /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(6730..6760) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6775..6796 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7084..7672) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7843..8703) /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(8704..8808) /gene="bla" /label="AmpR promoter" primer_bind 9282..9298 /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 cggaggcttc gagtgacccg 2341 ggtgccgagg agcgggaaga gttgctgggg cccactgctc agtggagcgt ggaggacgag 2401 gaggaggccg tccacgagca atgccagcat gagagagaca ggcagcttca ggcccaggac 2461 gaggagggag gcggccatgt ccccgagcgg ccgaagcagg agatgctcct cagcctgaag 2521 ccctcggagg cccctgaact ggatgaggac gagggctttg gcgactggtc ccagaggcca 2581 gagcagcggc agcagcacga gggggcgcag ggcgccttgg acagcggaga gcccccccag 2641 tgcaggagtc ctgaggggga gcaagaggac aggcccggcc tgcatgccta cgaaaaggag 2701 gacagtgatg aagtccacct ggaggagttg agtctgagca aggaggggcc aggcccagag 2761 gacactgtcc aggacaacct gggggccgca ggggctgagg aggaacagga ggagcaccag 2821 aaatgtcagc agcccaggac acccagcccc ttggtcttgg aggggaccat cgaacagagc 2881 tcgcctcccc tgagccctac caccaaactc atcgacagga ccgagtccct aaaccgctcc 2941 atagagaaga gtaacagtgt gaagaaatcc cagccagact tgcccatctc caagattgat 3001 cagtggctgg aacaatacac ccaggccatc gagaccgctg gccggacccc caagctagcc 3061 cgccaggcct ccatagagct gcccagcatg gctgtggcca gtaccaagag tcggtgggag 3121 acgggtgagg tacaggctca gtctgcggcc aagactccgt cctgcaagga tattgtggct 3181 ggagacatga gcaagaaaag cctctgggag cagaagggag gctccaagac ctcatcaaca 3241 attaagagca ccccatctgg gaagaggtat aagtttgtgg ccaccgggca tgggaagtat 3301 gagaaggtgc ttgtggaagg gggcccggct cccggatccg actacaaaga ccatgacggt 3361 gattataaag atcatgacat cgactacaag gatgacgatg acaagtagcg gccgcgaagg 3421 atctgcgatc gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag 3481 aagttggggg gaggggtcgg caattgaacg ggtgcctaga gaaggtggcg cggggtaaac 3541 tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat 3601 ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag 3661 ctgaagcttc gaggggctcg catctctcct tcacgcgccc gccgccctac ctgaggccgc 3721 catccacgcc ggttgagtcg cgttctgccg cctcccgcct gtggtgcctc ctgaactgcg 3781 tccgccgtct aggtaagttt aaagctcagg tcgagaccgg gcctttgtcc ggcgctccct 3841 tggagcctac ctagactcag ccggctctcc acgctttgcc tgaccctgct tgctcaactc 3901 tacgtctttg tttcgttttc tgttctgcgc cgttacagat ccaagctgtg accggcgcct 3961 acgctagacg ccaccatgga gagcgacgag agcggcctgc ccgccatgga gatcgagtgc 4021 cgcatcaccg gcaccctgaa cggcgtggag ttcgagctgg tgggcggcgg agagggcacc 4081 cccaagcagg gccgcatgac caacaagatg aagagcacca aaggcgccct gaccttcagc 4141 ccctacctgc tgagccacgt gatgggctac ggcttctacc acttcggcac ctaccccagc 4201 ggctacgaga accccttcct gcacgccatc aacaacggcg gctacaccaa cacccgcatc 4261 gagaagtacg aggacggcgg cgtgctgcac gtgagcttca gctaccgcta cgaggccggc 4321 cgcgtgatcg gcgacttcaa ggtggtgggc accggcttcc ccgaggacag cgtgatcttc 4381 accgacaaga tcatccgcag caacgccacc gtggagcacc tgcaccccat gggcgataac 4441 gtgctggtgg gcagcttcgc ccgcaccttc agcctgcgcg acggcggcta ctacagcttc 4501 gtggtggaca gccacatgca cttcaagagc gccatccacc ccagcatcct gcagaacggg 4561 ggccccatgt tcgccttccg ccgcgtggag gagctgcaca gcaacaccga gctgggcatc 4621 gtggagtacc agcacgcctt caagaccccc atcgccttcg ccagatcccg cgctcagtcg 4681 tccaattctg ccgtggacgg caccgccgga cccggctcca ccggatctcg cgagggcaga 4741 ggaagtcttc taacatgcgg tgacgtggag gagaatcccg gccctatgac cgagtacaag 4801 cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc 4861 gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg 4921 gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg 4981 gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg 5041 gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg 5101 cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg 5161 gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc 5221 cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc 5281 cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc 5341 gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgaaatca acctctggat 5401 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt 5461 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc 5521 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg 5581 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc 5641 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa 5701 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat 5761 tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc 5821 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt 5881 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag 5941 acgagtcgga tctccctttg ggccgcctcc ccgcctggta cctttaagac caatgactta 6001 caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat 6061 tcactcccaa cgaagataag atctgctttt tgcttgtact gggtctctct ggttagacca 6121 gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag 6181 cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag 6241 atccctcaga cccttttagt cagtgtggaa aatctctagc agtagtagtt catgtcatct 6301 tattattcag tatttataac ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt 6361 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc 6421 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt 6481 ctggctctag ctatcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc 6541 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc 6601 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct agacttttgc 6661 agagaccaaa ttcgtaatca tgtcatagct gtttcctgtg tgaaattgtt atccgctcac 6721 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt 6781 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc 6841 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg 6901 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt 6961 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa 7021 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc 7081 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag 7141 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt 7201 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg 7261 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg 7321 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg 7381 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac 7441 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg 7501 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt 7561 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg 7621 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc 7681 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt 7741 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt 7801 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag 7861 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt 7921 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc 7981 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc 8041 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg 8101 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac 8161 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg 8221 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc 8281 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact 8341 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc 8401 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat 8461 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc 8521 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac 8581 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa 8641 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact 8701 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg 8761 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg 8821 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag 8881 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca 8941 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc 9001 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc 9061 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag 9121 gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg 9181 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg 9241 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtgc 9301 caagctg //