LOCUS V035336 8947 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V035336 VERSION V035336 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..2973 /label="Slx9(NM_133998)" /note="Slx9(NM_133998)" /gene="Slx9" CDS 2980..3045 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3089..3300 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3313..3581 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 3610..3619 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 3640..4302 /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 4372..4425 /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 4426..5025 /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 5026..5614 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5688..5921 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 5993..6114 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6154..6289 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6322..6338) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6346..6362 /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(6370..6400) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6415..6436 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6724..7312) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7483..8343) /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(8344..8448) /gene="bla" /label="AmpR promoter" primer_bind 8922..8938 /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 ggaaagtgag ggcgctgcgg 2341 gctcgtgtgc accgggcggc cgtgagaccc gacggggatt ctgcaccggg ccctgtgccc 2401 cgcgccgtgg agccggcgct cccacaatcc ccggcgggcg gagcgggggc gaaggactgg 2461 acattcgttc acaatgacat ctttgccagg acacagatag accctagtgc gttggtgcag 2521 aggctggagc tggaccagag gagtgtggtg tccctaaaga gaggtgcaga gccaaaggcc 2581 attttgccta agaaggaaaa gctgaagctt cgccgtgagc gatggttaca gaaaattgaa 2641 gccataaagc tggcagagca gaagctcagg gaggaacgaa agcgcaaggc catggtggtg 2701 gtaggcgacc tgcaccccct gagggatgcc ctacctgagc tgcaggaact ggaggccggc 2761 cgccagcggc agcaagccag acgcagggtg accagcaagc cccggccagt ggagctcagc 2821 cgaatgacca cagtacagag gcagcagctt cttgaagaag aaaggacccg gtttcagaag 2881 ctgctggcca gcccaaccta cagagccagt cccctgctgg ccatcgggca gcagctggca 2941 caccagatgc agctggaagg tggcaaacag ctcggatccg actacaaaga ccatgacggt 3001 gattataaag atcatgacat cgactacaag gatgacgatg acaagtagcg gccgcgaagg 3061 atctgcgatc gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag 3121 aagttggggg gaggggtcgg caattgaacg ggtgcctaga gaaggtggcg cggggtaaac 3181 tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat 3241 ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag 3301 ctgaagcttc gaggggctcg catctctcct tcacgcgccc gccgccctac ctgaggccgc 3361 catccacgcc ggttgagtcg cgttctgccg cctcccgcct gtggtgcctc ctgaactgcg 3421 tccgccgtct aggtaagttt aaagctcagg tcgagaccgg gcctttgtcc ggcgctccct 3481 tggagcctac ctagactcag ccggctctcc acgctttgcc tgaccctgct tgctcaactc 3541 tacgtctttg tttcgttttc tgttctgcgc cgttacagat ccaagctgtg accggcgcct 3601 acgctagacg ccaccatgga gagcgacgag agcggcctgc ccgccatgga gatcgagtgc 3661 cgcatcaccg gcaccctgaa cggcgtggag ttcgagctgg tgggcggcgg agagggcacc 3721 cccaagcagg gccgcatgac caacaagatg aagagcacca aaggcgccct gaccttcagc 3781 ccctacctgc tgagccacgt gatgggctac ggcttctacc acttcggcac ctaccccagc 3841 ggctacgaga accccttcct gcacgccatc aacaacggcg gctacaccaa cacccgcatc 3901 gagaagtacg aggacggcgg cgtgctgcac gtgagcttca gctaccgcta cgaggccggc 3961 cgcgtgatcg gcgacttcaa ggtggtgggc accggcttcc ccgaggacag cgtgatcttc 4021 accgacaaga tcatccgcag caacgccacc gtggagcacc tgcaccccat gggcgataac 4081 gtgctggtgg gcagcttcgc ccgcaccttc agcctgcgcg acggcggcta ctacagcttc 4141 gtggtggaca gccacatgca cttcaagagc gccatccacc ccagcatcct gcagaacggg 4201 ggccccatgt tcgccttccg ccgcgtggag gagctgcaca gcaacaccga gctgggcatc 4261 gtggagtacc agcacgcctt caagaccccc atcgccttcg ccagatcccg cgctcagtcg 4321 tccaattctg ccgtggacgg caccgccgga cccggctcca ccggatctcg cgagggcaga 4381 ggaagtcttc taacatgcgg tgacgtggag gagaatcccg gccctatgac cgagtacaag 4441 cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc 4501 gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg 4561 gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg 4621 gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg 4681 gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg 4741 cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg 4801 gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc 4861 cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc 4921 cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc 4981 gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgaaatca acctctggat 5041 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt 5101 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc 5161 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg 5221 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc 5281 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa 5341 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat 5401 tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc 5461 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt 5521 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag 5581 acgagtcgga tctccctttg ggccgcctcc ccgcctggta cctttaagac caatgactta 5641 caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat 5701 tcactcccaa cgaagataag atctgctttt tgcttgtact gggtctctct ggttagacca 5761 gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag 5821 cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag 5881 atccctcaga cccttttagt cagtgtggaa aatctctagc agtagtagtt catgtcatct 5941 tattattcag tatttataac ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt 6001 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc 6061 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt 6121 ctggctctag ctatcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc 6181 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc 6241 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct agacttttgc 6301 agagaccaaa ttcgtaatca tgtcatagct gtttcctgtg tgaaattgtt atccgctcac 6361 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt 6421 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc 6481 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg 6541 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt 6601 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa 6661 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc 6721 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag 6781 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt 6841 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg 6901 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg 6961 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg 7021 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac 7081 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg 7141 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt 7201 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg 7261 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc 7321 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt 7381 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt 7441 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag 7501 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt 7561 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc 7621 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc 7681 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg 7741 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac 7801 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg 7861 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc 7921 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact 7981 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc 8041 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat 8101 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc 8161 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac 8221 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa 8281 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact 8341 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg 8401 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg 8461 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag 8521 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca 8581 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc 8641 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc 8701 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag 8761 gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg 8821 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg 8881 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtgc 8941 caagctg //