LOCUS V035252 9727 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V035252 VERSION V035252 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..3753 /label="Kmo(NM_133809)" /note="Kmo(NM_133809)" /gene="Kmo" CDS 3760..3825 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3869..4080 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4093..4361 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4390..4399 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4420..5082 /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 5152..5205 /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 5206..5805 /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 5806..6394 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6468..6701 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6773..6894 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6934..7069 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(7102..7118) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7126..7142 /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(7150..7180) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7195..7216 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7504..8092) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8263..9123) /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(9124..9228) /gene="bla" /label="AmpR promoter" primer_bind 9702..9718 /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 catcgtctga tactcagggg 2341 aaaagagtgg ctgttattgg cggtggtttg gttggagcac tgaatgcctg ctttcttgca 2401 aagaggaatt ttcaagttga tgtgtacgaa gctagggaag atattcgcgt ggctaaatct 2461 gcacgtggaa ggagcattaa cttggccctt tcttatagag gacggcaggc cttgaaagcc 2521 attggtctgg aagatcagat cgtttccaaa ggtgtgccca tgaaagccag aatgatccac 2581 tctctttcgg gaaagaagtc tgcaattccc tatgggaaca agtcacagta tatcctttca 2641 ataagcagag aaaacttaaa caaggacctg ctgactgccg tggagtccta tgccaatgcg 2701 aaggtgcact ttggccacaa gctgtcgaaa tgcattccgg aggaaggggt actcacagtg 2761 ctcggacctg acaaggttcc ccgagatgtc acatgtgacc ttgttgtagg gtgtgatgga 2821 gcctattcaa ctgtcagagc ccacctcatg aagaagcccc gctttgatta cactcagcaa 2881 tatatccctc atggatacat ggagttgaca attccaccta agaatgggga gtacgccatg 2941 gaacctaact gtcttcacat ttggcctaga aatgcctata tgatgatcgc ccttccaaac 3001 atggacaaat ctttcacatg caccttgttc atgccctttg aggagtttga aagacttcca 3061 acgcgcagcg atgtgctgga cttcttccag aagaactttc cagatgctat ccctctgatg 3121 ggagagcaag ccctcatgag agatttcttt ctgttgcctg cccagcccat gatatcagtg 3181 aagtgctctc ccttccacct gaagtcacac tgtgtgctga tgggagatgc cgctcatgcc 3241 attgtcccat tttttgggca aggaatgaat gcgggctttg aagactgctt ggtgtttgat 3301 gaattgatgg acaaattcaa taataatctt agtatgtgcc ttcctgaatt ctcaagattt 3361 aggatcccag atgaccatgc aatttcagac ctatctatgt acaattacat agagatgcga 3421 gcgcatgtca actctaggtg gttcctgttc caaaagctcc tggataaatt tcttcacgcg 3481 atcatgccct ctacctttat ccctctctat accatggtcg ccttcaccag aataagatac 3541 cacgaggcag tgctgcgttg gcattggcaa aaaaaggtga taaacagagg actctttgtc 3601 cttgggtccc tgatagccat tggaggcacc tacctacttg tgcaccatct gtccctgaga 3661 cctctggagt tcttgagaag acctgcctgg atgggaacca ctggctactg gactaggagt 3721 acagacattt ctctgcaagt tccatggagt tacggatccg actacaaaga ccatgacggt 3781 gattataaag atcatgacat cgactacaag gatgacgatg acaagtagcg gccgcgaagg 3841 atctgcgatc gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag 3901 aagttggggg gaggggtcgg caattgaacg ggtgcctaga gaaggtggcg cggggtaaac 3961 tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat 4021 ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag 4081 ctgaagcttc gaggggctcg catctctcct tcacgcgccc gccgccctac ctgaggccgc 4141 catccacgcc ggttgagtcg cgttctgccg cctcccgcct gtggtgcctc ctgaactgcg 4201 tccgccgtct aggtaagttt aaagctcagg tcgagaccgg gcctttgtcc ggcgctccct 4261 tggagcctac ctagactcag ccggctctcc acgctttgcc tgaccctgct tgctcaactc 4321 tacgtctttg tttcgttttc tgttctgcgc cgttacagat ccaagctgtg accggcgcct 4381 acgctagacg ccaccatgga gagcgacgag agcggcctgc ccgccatgga gatcgagtgc 4441 cgcatcaccg gcaccctgaa cggcgtggag ttcgagctgg tgggcggcgg agagggcacc 4501 cccaagcagg gccgcatgac caacaagatg aagagcacca aaggcgccct gaccttcagc 4561 ccctacctgc tgagccacgt gatgggctac ggcttctacc acttcggcac ctaccccagc 4621 ggctacgaga accccttcct gcacgccatc aacaacggcg gctacaccaa cacccgcatc 4681 gagaagtacg aggacggcgg cgtgctgcac gtgagcttca gctaccgcta cgaggccggc 4741 cgcgtgatcg gcgacttcaa ggtggtgggc accggcttcc ccgaggacag cgtgatcttc 4801 accgacaaga tcatccgcag caacgccacc gtggagcacc tgcaccccat gggcgataac 4861 gtgctggtgg gcagcttcgc ccgcaccttc agcctgcgcg acggcggcta ctacagcttc 4921 gtggtggaca gccacatgca cttcaagagc gccatccacc ccagcatcct gcagaacggg 4981 ggccccatgt tcgccttccg ccgcgtggag gagctgcaca gcaacaccga gctgggcatc 5041 gtggagtacc agcacgcctt caagaccccc atcgccttcg ccagatcccg cgctcagtcg 5101 tccaattctg ccgtggacgg caccgccgga cccggctcca ccggatctcg cgagggcaga 5161 ggaagtcttc taacatgcgg tgacgtggag gagaatcccg gccctatgac cgagtacaag 5221 cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc 5281 gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg 5341 gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg 5401 gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg 5461 gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg 5521 cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg 5581 gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc 5641 cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc 5701 cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc 5761 gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgaaatca acctctggat 5821 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt 5881 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc 5941 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg 6001 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc 6061 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa 6121 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat 6181 tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc 6241 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt 6301 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag 6361 acgagtcgga tctccctttg ggccgcctcc ccgcctggta cctttaagac caatgactta 6421 caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat 6481 tcactcccaa cgaagataag atctgctttt tgcttgtact gggtctctct ggttagacca 6541 gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag 6601 cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag 6661 atccctcaga cccttttagt cagtgtggaa aatctctagc agtagtagtt catgtcatct 6721 tattattcag tatttataac ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt 6781 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc 6841 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt 6901 ctggctctag ctatcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc 6961 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc 7021 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct agacttttgc 7081 agagaccaaa ttcgtaatca tgtcatagct gtttcctgtg tgaaattgtt atccgctcac 7141 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt 7201 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc 7261 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg 7321 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt 7381 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa 7441 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc 7501 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag 7561 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt 7621 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg 7681 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg 7741 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg 7801 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac 7861 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg 7921 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt 7981 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg 8041 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc 8101 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt 8161 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt 8221 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag 8281 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt 8341 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc 8401 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc 8461 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg 8521 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac 8581 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg 8641 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc 8701 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact 8761 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc 8821 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat 8881 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc 8941 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac 9001 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa 9061 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact 9121 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg 9181 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg 9241 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag 9301 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca 9361 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc 9421 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc 9481 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag 9541 gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg 9601 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg 9661 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtgc 9721 caagctg //