LOCUS V043770 7927 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V043770 VERSION V043770 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..2781 /label="Ier3(NM_133662)" /note="Ier3(NM_133662)" /gene="Ier3" CDS 2788..2853 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 2897..3108 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3121..3389 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 3416..4015 /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 4022..4610 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 4684..4917 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 4989..5110 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 5129..5264 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(5302..5318) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5326..5342 /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(5350..5380) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5395..5416 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5704..6292) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6463..7323) /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(7324..7428) /gene="bla" /label="AmpR promoter" primer_bind 7902..7918 /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 catgtgccac tcgcgcaacc atctccacac catgactggc 2341 ctgagggccc cttctccagc tccctccacc ggcccggaac tccggcgggg ctctggtccc 2401 gaaattttca ccttcgaccc tctcccggag cgggccgtgg tgtccaccgc gcgtttgaac 2461 acttctcgcg ggcaccgaaa acgcagccga agggtgctct accctcgagt ggtccggcgc 2521 cagctaccaa ccgaggaacc caacattgcc aagagggtcc tctttctcct gttcgccatc 2581 atcttctgcc agattttgat ggctgaagag ggtgtgtcgc agcccctggc tccggaggat 2641 gctaccagcg ccgtgacacc tgagcccatt tctgcgccca ttactgcgcc cccggtcctc 2701 gagcctttga acctgacctc ggagtcctcg gactatgcgc tggatcttaa agcttttctc 2761 cagcaacatc cggcggcctt cggatccgac tacaaagacc atgacggtga ttataaagat 2821 catgacatcg actacaagga tgacgatgac aagtagcggc cgcgaaggat ctgcgatcgc 2881 tccggtgccc gtcagtgggc agagcgcaca tcgcccacag tccccgagaa gttgggggga 2941 ggggtcggca attgaacggg tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat 3001 gtcgtgtact ggctccgcct ttttcccgag ggtgggggag aaccgtatat aagtgcagta 3061 gtcgccgtga acgttctttt tcgcaacggg tttgccgcca gaacacagct gaagcttcga 3121 ggggctcgca tctctccttc acgcgcccgc cgccctacct gaggccgcca tccacgccgg 3181 ttgagtcgcg ttctgccgcc tcccgcctgt ggtgcctcct gaactgcgtc cgccgtctag 3241 gtaagtttaa agctcaggtc gagaccgggc ctttgtccgg cgctcccttg gagcctacct 3301 agactcagcc ggctctccac gctttgcctg accctgcttg ctcaactcta cgtctttgtt 3361 tcgttttctg ttctgcgccg ttacagatcc aagctgtgac cggcgcctac gctagatgac 3421 cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtcccccggg ccgtacgcac 3481 cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgacc cggaccgcca 3541 catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg 3601 caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt 3661 cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg 3721 gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc 3781 gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc 3841 cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac 3901 ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt 3961 cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgagtcga 4021 caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc 4081 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg 4141 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt 4201 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac 4261 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc 4321 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct 4381 gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct 4441 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct 4501 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct 4561 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggtacctt 4621 taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg 4681 gactggaagg gctaattcac tcccaacgaa aataagatct gctttttgct tgtactgggt 4741 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc 4801 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg 4861 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta 4921 gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga 4981 gtgagaggaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa 5041 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca 5101 atgtatctta tcatgtctgg ctctagctat cccgccccta actccgccca gttccgccca 5161 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc 5221 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctagact tttgcagaga 5281 cggcccaaat tcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac 5341 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt 5401 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc 5461 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg 5521 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt 5581 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa 5641 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc 5701 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag 5761 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt 5821 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg 5881 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg 5941 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg 6001 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac 6061 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg 6121 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt 6181 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg 6241 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc 6301 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt 6361 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt 6421 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag 6481 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt 6541 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc 6601 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc 6661 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg 6721 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac 6781 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg 6841 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc 6901 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact 6961 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc 7021 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat 7081 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc 7141 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac 7201 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa 7261 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact 7321 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg 7381 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg 7441 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag 7501 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca 7561 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc 7621 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc 7681 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag 7741 gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg 7801 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg 7861 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtgc 7921 caagctg //