LOCUS V043309 9850 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V043309 VERSION V043309 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..3876 /label="USP3(NM_006537)" /note="USP3(NM_006537)" /gene="USP3" CDS 3883..3948 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3992..4203 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4216..4484 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4513..4522 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4543..5205 /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 5275..5328 /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 5329..5928 /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 5929..6517 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6591..6824 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6896..7017 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 7057..7192 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(7225..7241) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7249..7265 /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(7273..7303) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7318..7339 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7627..8215) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8386..9246) /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(9247..9351) /gene="bla" /label="AmpR promoter" primer_bind 9825..9841 /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 agtgtccaca cctgagctcc 2341 agcgtctgca ttgctccgga ctcagccaag ttccccaacg gctccccgtc gtcctggtgc 2401 tgcagcgtgt gccggtccaa caaaagccct tgggtctgtt tgacttgttc aagtgtccac 2461 tgtggaaggt atgtgaatgg ccatgcaaaa aaacattatg aagatgcaca agtaccttta 2521 accaaccata agaaatcaga aaagcaagat aaagttcagc acacagtatg tatggattgc 2581 agtagctaca gtacatactg ttatcgctgt gatgattttg tggttaatga caccaagctg 2641 ggactggtac agaaagtcag agaacactta cagaacttgg aaaactcagc tttcacagct 2701 gacaggcata agaaaagaaa acttttggaa aactcaacac taaacagcaa gttattaaaa 2761 gtaaatggaa gcaccactgc catttgtgcc acaggccttc ggaatttggg gaacacatgt 2821 ttcatgaatg ccatccttca gtcactcagt aacattgagc agttttgctg ttatttcaaa 2881 gaactgcccg ccgtggagtt aaggaatggg aaaacagcag gaaggcggac ataccacacc 2941 aggagccaag gggataacaa tgtgtctttg gtagaagagt ttagaaagac actctgtgct 3001 ttatggcaag gcagccagac tgcatttagc ccagagtcct tattttatgt tgtttggaag 3061 attatgccaa actttagggg ctatcaacag caggacgccc atgaattcat gcgctacctt 3121 ttggaccacc tacacttgga acttcagggc ggtttcaacg gtgtttcccg ctcagcaatt 3181 ctgcaggaga attctactct gtctgcaagt aacaagtgtt gcataaatgg agcatctact 3241 gttgtcacgg ctatattcgg aggcattctc caaaatgagg ttaactgcct catatgtggg 3301 acagaatcta gaaagtttga tccattccta gacctttcat tagatattcc aagtcagttc 3361 agaagtaagc gctctaagaa tcaagaaaat ggaccagttt gttcgttacg agattgtctt 3421 cgcagtttta ccgacttaga agaacttgat gagacagagt tatatatgtg ccataaatgc 3481 aaaaagaaac aaaagtccac aaaaaagttt tggattcaaa aactacccaa ggtgctatgc 3541 ttacatttga aaagatttca ttggacagca tatttaagaa ataaagttga tacatacgta 3601 gaatttccac tgagaggcct agacatgaaa tgctacttac tagagcctga gaacagtggc 3661 ccggagagct gcctgtatga cctcgccgct gtggtggtgc accatggttc cggggttggt 3721 tctggacatt acacagcata cgcaactcac gaaggccgct ggttccactt caatgacagt 3781 actgtaacac tgactgacga ggagactgtg gtgaaggcga aggcctacat ccttttctac 3841 gtggaacacc aggccaaagc tggatcggat aaacttggat ccgactacaa agaccatgac 3901 ggtgattata aagatcatga catcgactac aaggatgacg atgacaagta gcggccgcga 3961 aggatctgcg atcgctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc 4021 gagaagttgg ggggaggggt cggcaattga acgggtgcct agagaaggtg gcgcggggta 4081 aactgggaaa gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg 4141 tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca 4201 cagctgaagc ttcgaggggc tcgcatctct ccttcacgcg cccgccgccc tacctgaggc 4261 cgccatccac gccggttgag tcgcgttctg ccgcctcccg cctgtggtgc ctcctgaact 4321 gcgtccgccg tctaggtaag tttaaagctc aggtcgagac cgggcctttg tccggcgctc 4381 ccttggagcc tacctagact cagccggctc tccacgcttt gcctgaccct gcttgctcaa 4441 ctctacgtct ttgtttcgtt ttctgttctg cgccgttaca gatccaagct gtgaccggcg 4501 cctacgctag acgccaccat ggagagcgac gagagcggcc tgcccgccat ggagatcgag 4561 tgccgcatca ccggcaccct gaacggcgtg gagttcgagc tggtgggcgg cggagagggc 4621 acccccaagc agggccgcat gaccaacaag atgaagagca ccaaaggcgc cctgaccttc 4681 agcccctacc tgctgagcca cgtgatgggc tacggcttct accacttcgg cacctacccc 4741 agcggctacg agaacccctt cctgcacgcc atcaacaacg gcggctacac caacacccgc 4801 atcgagaagt acgaggacgg cggcgtgctg cacgtgagct tcagctaccg ctacgaggcc 4861 ggccgcgtga tcggcgactt caaggtggtg ggcaccggct tccccgagga cagcgtgatc 4921 ttcaccgaca agatcatccg cagcaacgcc accgtggagc acctgcaccc catgggcgat 4981 aacgtgctgg tgggcagctt cgcccgcacc ttcagcctgc gcgacggcgg ctactacagc 5041 ttcgtggtgg acagccacat gcacttcaag agcgccatcc accccagcat cctgcagaac 5101 gggggcccca tgttcgcctt ccgccgcgtg gaggagctgc acagcaacac cgagctgggc 5161 atcgtggagt accagcacgc cttcaagacc cccatcgcct tcgccagatc ccgcgctcag 5221 tcgtccaatt ctgccgtgga cggcaccgcc ggacccggct ccaccggatc tcgcgagggc 5281 agaggaagtc ttctaacatg cggtgacgtg gaggagaatc ccggccctat gaccgagtac 5341 aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg caccctcgcc 5401 gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg ccacatcgag 5461 cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg 5521 tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg 5581 ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc 5641 gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc 5701 ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg 5761 ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg 5821 ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg 5881 cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgaaa tcaacctctg 5941 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta 6001 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt 6061 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc 6121 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt 6181 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg 6241 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac 6301 aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc 6361 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac 6421 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct 6481 cagacgagtc ggatctccct ttgggccgcc tccccgcctg gtacctttaa gaccaatgac 6541 ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct 6601 aattcactcc caacgaagat aagatctgct ttttgcttgt actgggtctc tctggttaga 6661 ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata 6721 aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta 6781 gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtagta gttcatgtca 6841 tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggaactt 6901 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa 6961 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca 7021 tgtctggctc tagctatccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt 7081 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc 7141 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctagacttt 7201 tgcagagacc aaattcgtaa tcatgtcata gctgtttcct gtgtgaaatt gttatccgct 7261 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg 7321 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct 7381 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg 7441 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc 7501 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg 7561 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct 7621 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca 7681 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct 7741 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc 7801 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt 7861 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc 7921 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc 7981 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg 8041 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc 8101 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag 8161 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga 8221 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat 8281 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag 8341 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat 8401 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc 8461 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat 8521 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag 8581 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg 8641 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc 8701 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca 8761 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg 8821 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc 8881 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta 8941 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc 9001 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg 9061 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc 9121 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc 9181 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat 9241 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag 9301 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc 9361 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa 9421 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg 9481 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca 9541 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc 9601 atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt 9661 aaggagaaaa taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg 9721 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag 9781 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag 9841 tgccaagctg //