LOCUS V047226 8626 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V047226 VERSION V047226 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..3480 /codon_start=1 /gene="Homo sapiens TP53" /note="responds to cellular stresses to regulate the expression of target genes" /product="human tumor suppressor protein that is often mutated in cancers" /transl_table=1 /translation="MEEPQSDPSVEPPLSQETFSDLWKLLPENNVLSPLPSQAMDDLML SPDDIEQWFTEDPGPDEAPRMPEAAPPVAPAPAAPTPAAPAPAPSWPLSSSVPSQKTYQ GSYGFRLGFLHSGTAKSVTCTYSPALNKMFCQLAKTCPVQLWVDSTPPPGTRVRAMAIY KQSQHMTEVVRHCPHHERCSDSDGLAPPQHLIRVEGNLRVEYLDDRNTFRHSVVVPYEP PEVGSDCTTIHYNYMCNSSCMGGMNRRPILTIITLEDSSGNLLGRNSFEVRVCACPGRD RRTEEENLRKKGEPHHELPPGSTKRALPNNTSSSPQPKKKPLDGEYFTLQIRGRERFEM FRELNEALELKDAQAGKEPGGSRAHSSHLKSKKGQSTSRHKKLMFKTEGPDSD" /label="p53" CDS 3487..3552 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3596..3807 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3820..4088 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 4115..4714 /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 4721..5309 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5383..5616 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 5688..5809 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 5828..5963 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6001..6017) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6025..6041 /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(6049..6079) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6094..6115 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6403..6991) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7162..8022) /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(8023..8127) /gene="bla" /label="AmpR promoter" primer_bind 8601..8617 /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 catggaggag ccgcagtcag atcctagcgt cgagccccct 2341 ctgagtcagg aaacattttc agacctatgg aaactacttc ctgaaaacaa cgttctgtcc 2401 cccttgccgt cccaagcaat ggatgatttg atgctgtccc cggacgatat tgaacaatgg 2461 ttcactgaag acccaggtcc agatgaagct cccagaatgc cagaggctgc tccccccgtg 2521 gcccctgcac cagcagctcc tacaccggcg gcccctgcac cagccccctc ctggcccctg 2581 tcatcttctg tcccttccca gaaaacctac cagggcagct acggtttccg tctgggcttc 2641 ttgcattctg ggacagccaa gtctgtgact tgcacgtact cccctgccct caacaagatg 2701 ttttgccaac tggccaagac ctgccctgtg cagctgtggg ttgattccac acccccgccc 2761 ggcacccgcg tccgcgccat ggccatctac aagcagtcac agcacatgac ggaggttgtg 2821 aggcactgcc cccaccatga gcgctgctca gatagcgatg gtctggcccc tcctcagcat 2881 cttatccgag tggaaggaaa tttgcgtgtg gagtatttgg atgacagaaa cacttttcga 2941 catagtgtgg tggtgcccta tgagccgcct gaggttggct ctgactgtac caccatccac 3001 tacaactaca tgtgtaacag ttcctgcatg ggcggcatga accggaggcc catcctcacc 3061 atcatcacac tggaagactc cagtggtaat ctactgggac ggaacagctt tgaggtgcgt 3121 gtttgtgcct gtcctgggag agaccggcgc acagaggaag agaatctccg caagaaaggg 3181 gagcctcacc acgagctgcc cccagggagc actaagcgag cactgcccaa caacaccagc 3241 tcctctcccc agccaaagaa gaaaccactg gatggagaat atttcaccct tcagatccgt 3301 gggcgtgagc gcttcgagat gttccgagag ctgaatgagg ccttggaact caaggatgcc 3361 caggctggga aggagccagg ggggagcagg gctcactcca gccacctgaa gtccaaaaag 3421 ggtcagtcta cctcccgcca taaaaaactc atgttcaaga cagaagggcc tgactcagac 3481 ggatccgact acaaagacca tgacggtgat tataaagatc atgacatcga ctacaaggat 3541 gacgatgaca agtagcggcc gcgaaggatc tgcgatcgct ccggtgcccg tcagtgggca 3601 gagcgcacat cgcccacagt ccccgagaag ttggggggag gggtcggcaa ttgaacgggt 3661 gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt 3721 tttcccgagg gtgggggaga accgtatata agtgcagtag tcgccgtgaa cgttcttttt 3781 cgcaacgggt ttgccgccag aacacagctg aagcttcgag gggctcgcat ctctccttca 3841 cgcgcccgcc gccctacctg aggccgccat ccacgccggt tgagtcgcgt tctgccgcct 3901 cccgcctgtg gtgcctcctg aactgcgtcc gccgtctagg taagtttaaa gctcaggtcg 3961 agaccgggcc tttgtccggc gctcccttgg agcctaccta gactcagccg gctctccacg 4021 ctttgcctga ccctgcttgc tcaactctac gtctttgttt cgttttctgt tctgcgccgt 4081 tacagatcca agctgtgacc ggcgcctacg ctagatgacc gagtacaagc ccacggtgcg 4141 cctcgccacc cgcgacgacg tcccccgggc cgtacgcacc ctcgccgccg cgttcgccga 4201 ctaccccgcc acgcgccaca ccgtcgaccc ggaccgccac atcgagcggg tcaccgagct 4261 gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga 4321 cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc 4381 cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat 4441 ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg 4501 cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga 4561 ggcggccgag cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc 4621 cttctacgag cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg 4681 cacctggtgc atgacccgca agcccggtgc ctgagtcgac aatcaacctc tggattacaa 4741 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata 4801 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc 4861 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg 4921 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac 4981 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat 5041 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt 5101 ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat 5161 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc 5221 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag 5281 tcggatctcc ctttgggccg cctccccgcc tggtaccttt aagaccaatg acttacaagg 5341 cagctgtaga tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact 5401 cccaacgaaa ataagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct 5461 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc 5521 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc 5581 tcagaccctt ttagtcagtg tggaaaatct ctagcagtag tagttcatgt catcttatta 5641 ttcagtattt ataacttgca aagaaatgaa tatcagagag tgagaggaac ttgtttattg 5701 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt 5761 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctggc 5821 tctagctatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact 5881 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta 5941 gtgaggaggc ttttttggag gcctagactt ttgcagagac ggcccaaatt cgtaatcatg 6001 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc 6061 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc 6121 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat 6181 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac 6241 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt 6301 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca 6361 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc 6421 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact 6481 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct 6541 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag 6601 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca 6661 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa 6721 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc 6781 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag 6841 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg 6901 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca 6961 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc 7021 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag 7081 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata 7141 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat 7201 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg 7261 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc 7321 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc 7381 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc 7441 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc 7501 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc 7561 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa 7621 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat 7681 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata 7741 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca 7801 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag 7861 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc 7921 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc 7981 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata 8041 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta 8101 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta 8161 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg 8221 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt 8281 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg 8341 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt 8401 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg 8461 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct 8521 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg 8581 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctg //