LOCUS V030402 9007 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V030402 VERSION V030402 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..1920 /note="central polypurine tract and central termination sequence of HIV-1" /label="cPPT/CTS" promoter 2010..2213 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 2302..3861 /label="USP3(NM_006537)" /note="USP3(NM_006537)" /gene="USP3" CDS 3868..3933 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3977..4188 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4201..4469 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 4496..5095 /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 5102..5690 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5764..5997 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6069..6190 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6209..6344 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6382..6398) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6406..6422 /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(6430..6460) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6475..6496 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6784..7372) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7543..8403) /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(8404..8508) /gene="bla" /label="AmpR promoter" primer_bind 8982..8998 /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 attacaaaat tcaaaatttt 1921 atcgatacta gtattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca 1981 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc 2041 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga 2101 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat 2161 tgacgcaaat gggcggtagg cgtgtacggt gggaggttta tataagcaga gctcgtttag 2221 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagattct 2281 agagctagcg aattcgccac catggagtgt ccacacctga gctccagcgt ctgcattgct 2341 ccggactcag ccaagttccc caacggctcc ccgtcgtcct ggtgctgcag cgtgtgccgg 2401 tccaacaaaa gcccttgggt ctgtttgact tgttcaagtg tccactgtgg aaggtatgtg 2461 aatggccatg caaaaaaaca ttatgaagat gcacaagtac ctttaaccaa ccataagaaa 2521 tcagaaaagc aagataaagt tcagcacaca gtatgtatgg attgcagtag ctacagtaca 2581 tactgttatc gctgtgatga ttttgtggtt aatgacacca agctgggact ggtacagaaa 2641 gtcagagaac acttacagaa cttggaaaac tcagctttca cagctgacag gcataagaaa 2701 agaaaacttt tggaaaactc aacactaaac agcaagttat taaaagtaaa tggaagcacc 2761 actgccattt gtgccacagg ccttcggaat ttggggaaca catgtttcat gaatgccatc 2821 cttcagtcac tcagtaacat tgagcagttt tgctgttatt tcaaagaact gcccgccgtg 2881 gagttaagga atgggaaaac agcaggaagg cggacatacc acaccaggag ccaaggggat 2941 aacaatgtgt ctttggtaga agagtttaga aagacactct gtgctttatg gcaaggcagc 3001 cagactgcat ttagcccaga gtccttattt tatgttgttt ggaagattat gccaaacttt 3061 aggggctatc aacagcagga cgcccatgaa ttcatgcgct accttttgga ccacctacac 3121 ttggaacttc agggcggttt caacggtgtt tcccgctcag caattctgca ggagaattct 3181 actctgtctg caagtaacaa gtgttgcata aatggagcat ctactgttgt cacggctata 3241 ttcggaggca ttctccaaaa tgaggttaac tgcctcatat gtgggacaga atctagaaag 3301 tttgatccat tcctagacct ttcattagat attccaagtc agttcagaag taagcgctct 3361 aagaatcaag aaaatggacc agtttgttcg ttacgagatt gtcttcgcag ttttaccgac 3421 ttagaagaac ttgatgagac agagttatat atgtgccata aatgcaaaaa gaaacaaaag 3481 tccacaaaaa agttttggat tcaaaaacta cccaaggtgc tatgcttaca tttgaaaaga 3541 tttcattgga cagcatattt aagaaataaa gttgatacat acgtagaatt tccactgaga 3601 ggcctagaca tgaaatgcta cttactagag cctgagaaca gtggcccgga gagctgcctg 3661 tatgacctcg ccgctgtggt ggtgcaccat ggttccgggg ttggttctgg acattacaca 3721 gcatacgcaa ctcacgaagg ccgctggttc cacttcaatg acagtactgt aacactgact 3781 gacgaggaga ctgtggtgaa ggcgaaggcc tacatccttt tctacgtgga acaccaggcc 3841 aaagctggat cggataaact tggatccgac tacaaagacc atgacggtga ttataaagat 3901 catgacatcg actacaagga tgacgatgac aagtagcggc cgcgaaggat ctgcgatcgc 3961 tccggtgccc gtcagtgggc agagcgcaca tcgcccacag tccccgagaa gttgggggga 4021 ggggtcggca attgaacggg tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat 4081 gtcgtgtact ggctccgcct ttttcccgag ggtgggggag aaccgtatat aagtgcagta 4141 gtcgccgtga acgttctttt tcgcaacggg tttgccgcca gaacacagct gaagcttcga 4201 ggggctcgca tctctccttc acgcgcccgc cgccctacct gaggccgcca tccacgccgg 4261 ttgagtcgcg ttctgccgcc tcccgcctgt ggtgcctcct gaactgcgtc cgccgtctag 4321 gtaagtttaa agctcaggtc gagaccgggc ctttgtccgg cgctcccttg gagcctacct 4381 agactcagcc ggctctccac gctttgcctg accctgcttg ctcaactcta cgtctttgtt 4441 tcgttttctg ttctgcgccg ttacagatcc aagctgtgac cggcgcctac gctagatgac 4501 cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtcccccggg ccgtacgcac 4561 cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgacc cggaccgcca 4621 catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg 4681 caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt 4741 cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg 4801 gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc 4861 gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc 4921 cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac 4981 ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt 5041 cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgagtcga 5101 caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc 5161 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg 5221 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt 5281 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac 5341 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc 5401 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct 5461 gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct 5521 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct 5581 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct 5641 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggtacctt 5701 taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg 5761 gactggaagg gctaattcac tcccaacgaa aataagatct gctttttgct tgtactgggt 5821 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc 5881 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg 5941 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta 6001 gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga 6061 gtgagaggaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa 6121 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca 6181 atgtatctta tcatgtctgg ctctagctat cccgccccta actccgccca gttccgccca 6241 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc 6301 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctagact tttgcagaga 6361 cggcccaaat tcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac 6421 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt 6481 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc 6541 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg 6601 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt 6661 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa 6721 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc 6781 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag 6841 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt 6901 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg 6961 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg 7021 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg 7081 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac 7141 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg 7201 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt 7261 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg 7321 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc 7381 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt 7441 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt 7501 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag 7561 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt 7621 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc 7681 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc 7741 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg 7801 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac 7861 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg 7921 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc 7981 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact 8041 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc 8101 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat 8161 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc 8221 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac 8281 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa 8341 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact 8401 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg 8461 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg 8521 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag 8581 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca 8641 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc 8701 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc 8761 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag 8821 gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg 8881 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg 8941 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtgc 9001 caagctg //