LOCUS V040694 9367 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V040694 VERSION V040694 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..3393 /label="PDHB(NM_000925)" /note="PDHB(NM_000925)" /gene="PDHB" CDS 3400..3465 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3509..3720 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3733..4001 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4030..4039 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4060..4722 /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 4792..4845 /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 4846..5445 /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 5446..6034 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6108..6341 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6413..6534 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6574..6709 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6742..6758) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6766..6782 /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(6790..6820) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6835..6856 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7144..7732) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7903..8763) /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(8764..8868) /gene="bla" /label="AmpR promoter" primer_bind 9342..9358 /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 cggcggtgtc tggcttggtg 2341 cggagacccc ttcgggaggt ctccgggctg ctgaagaggc gctttcactg gaccgcgccg 2401 gctgcgctgc aggtgacagt tcgtgatgct ataaatcagg gtatggatga ggagctggaa 2461 agagatgaga aggtatttct gcttggagaa gaagttgccc agtatgatgg ggcatacaag 2521 gttagtcgag ggctgtggaa gaaatatgga gacaagagga ttattgacac tcccatatca 2581 gagatgggct ttgctggaat tgctgtaggt gcagctatgg ctgggttgcg gcccatttgt 2641 gaatttatga ccttcaattt ctccatgcaa gccattgacc aggttataaa ctcagctgcc 2701 aagacctact acatgtctgg tggccttcag cctgtgccta tagtcttcag agggcccaat 2761 ggtgcctcag caggtgtagc tgcccagcac tcacagtgct ttgctgcctg gtatgggcac 2821 tgcccaggct taaaggtggt cagtccctgg aattcagagg atgctaaagg acttattaaa 2881 tcagccattc gggataacaa tccagtggtg gtgctagaga atgaattgat gtatggggtt 2941 ccttttgaat ttcctccgga agctcagtca aaagattttc tgattcctat tggaaaagcc 3001 aaaatagaaa ggcaaggaac acatataact gtggtttccc attcaagacc tgtgggccac 3061 tgcttagaag ctgcagcagt gctatctaaa gaaggagttg aatgtgaggt gataaatatg 3121 cgtaccatta gaccaatgga catggaaacc atagaagcca gtgtcatgaa gacaaatcat 3181 cttgtaactg tggaaggagg ctggccacag tttggagtag gagctgaaat ctgtgccagg 3241 atcatggaag gtcctgcgtt caatttcctg gatgctcctg ctgttcgtgt cactggtgct 3301 gatgtcccta tgccttatgc aaagattcta gaggacaact ctatacctca ggtcaaagac 3361 atcatatttg caataaagaa aacattaaat attggatccg actacaaaga ccatgacggt 3421 gattataaag atcatgacat cgactacaag gatgacgatg acaagtagcg gccgcgaagg 3481 atctgcgatc gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag 3541 aagttggggg gaggggtcgg caattgaacg ggtgcctaga gaaggtggcg cggggtaaac 3601 tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat 3661 ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag 3721 ctgaagcttc gaggggctcg catctctcct tcacgcgccc gccgccctac ctgaggccgc 3781 catccacgcc ggttgagtcg cgttctgccg cctcccgcct gtggtgcctc ctgaactgcg 3841 tccgccgtct aggtaagttt aaagctcagg tcgagaccgg gcctttgtcc ggcgctccct 3901 tggagcctac ctagactcag ccggctctcc acgctttgcc tgaccctgct tgctcaactc 3961 tacgtctttg tttcgttttc tgttctgcgc cgttacagat ccaagctgtg accggcgcct 4021 acgctagacg ccaccatgga gagcgacgag agcggcctgc ccgccatgga gatcgagtgc 4081 cgcatcaccg gcaccctgaa cggcgtggag ttcgagctgg tgggcggcgg agagggcacc 4141 cccaagcagg gccgcatgac caacaagatg aagagcacca aaggcgccct gaccttcagc 4201 ccctacctgc tgagccacgt gatgggctac ggcttctacc acttcggcac ctaccccagc 4261 ggctacgaga accccttcct gcacgccatc aacaacggcg gctacaccaa cacccgcatc 4321 gagaagtacg aggacggcgg cgtgctgcac gtgagcttca gctaccgcta cgaggccggc 4381 cgcgtgatcg gcgacttcaa ggtggtgggc accggcttcc ccgaggacag cgtgatcttc 4441 accgacaaga tcatccgcag caacgccacc gtggagcacc tgcaccccat gggcgataac 4501 gtgctggtgg gcagcttcgc ccgcaccttc agcctgcgcg acggcggcta ctacagcttc 4561 gtggtggaca gccacatgca cttcaagagc gccatccacc ccagcatcct gcagaacggg 4621 ggccccatgt tcgccttccg ccgcgtggag gagctgcaca gcaacaccga gctgggcatc 4681 gtggagtacc agcacgcctt caagaccccc atcgccttcg ccagatcccg cgctcagtcg 4741 tccaattctg ccgtggacgg caccgccgga cccggctcca ccggatctcg cgagggcaga 4801 ggaagtcttc taacatgcgg tgacgtggag gagaatcccg gccctatgac cgagtacaag 4861 cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc 4921 gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg 4981 gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg 5041 gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg 5101 gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg 5161 cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg 5221 gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc 5281 cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc 5341 cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc 5401 gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgaaatca acctctggat 5461 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt 5521 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc 5581 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg 5641 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc 5701 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa 5761 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat 5821 tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc 5881 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt 5941 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag 6001 acgagtcgga tctccctttg ggccgcctcc ccgcctggta cctttaagac caatgactta 6061 caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat 6121 tcactcccaa cgaagataag atctgctttt tgcttgtact gggtctctct ggttagacca 6181 gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag 6241 cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag 6301 atccctcaga cccttttagt cagtgtggaa aatctctagc agtagtagtt catgtcatct 6361 tattattcag tatttataac ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt 6421 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc 6481 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt 6541 ctggctctag ctatcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc 6601 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc 6661 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct agacttttgc 6721 agagaccaaa ttcgtaatca tgtcatagct gtttcctgtg tgaaattgtt atccgctcac 6781 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt 6841 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc 6901 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg 6961 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt 7021 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa 7081 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc 7141 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag 7201 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt 7261 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg 7321 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg 7381 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg 7441 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac 7501 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg 7561 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt 7621 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg 7681 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc 7741 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt 7801 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt 7861 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag 7921 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt 7981 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc 8041 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc 8101 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg 8161 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac 8221 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg 8281 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc 8341 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact 8401 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc 8461 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat 8521 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc 8581 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac 8641 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa 8701 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact 8761 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg 8821 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg 8881 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag 8941 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca 9001 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc 9061 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc 9121 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag 9181 gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg 9241 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg 9301 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtgc 9361 caagctg //