LOCUS V046450 8467 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V046450 VERSION V046450 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..3321 /label="DUSP12(NM_007240)" /note="DUSP12(NM_007240)" /gene="DUSP12" CDS 3328..3393 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3437..3648 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3661..3929 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 3956..4555 /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 4562..5150 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5224..5457 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 5529..5650 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 5669..5804 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(5842..5858) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5866..5882 /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(5890..5920) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5935..5956 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6244..6832) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7003..7863) /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(7864..7968) /gene="bla" /label="AmpR promoter" primer_bind 8442..8458 /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 catgttggag gctccgggcc cgagtgatgg ctgcgagctc 2341 agcaacccca gcgccagcag agtcagctgt gccgggcaga tgctggaagt gcagccagga 2401 ttgtatttcg gtggggccgc ggccgtcgcg gagccagatc acctgaggga agcgggcatc 2461 acggccgtgc taacagtgga ctcggaggag cccagcttca aggcggggcc tggggtcgag 2521 gatctatggc gcctcttcgt gccagcgctg gacaaacccg agacggacct actcagccat 2581 ctggaccggt gcgtggcctt catcggtcag gcccgcgctg agggccgtgc ggtgttggtg 2641 cactgtcatg caggagtcag tcgaagtgtg gccataataa ctgcttttct catgaagact 2701 gaccaacttc cctttgaaaa agcctatgaa aagctccaga ttctcaaacc agaggctaag 2761 atgaatgagg ggtttgagtg gcaactgaaa ttataccagg caatgggata cgaagtggat 2821 acctctagtg caatttataa gcaatatcgt ttacaaaagg ttacagagaa gtatccagaa 2881 ttgcagaatt tacctcaaga actctttgct gttgacccaa ctaccgtttc acaaggattg 2941 aaagatgagg ttctctacaa gtgtagaaag tgcaggcgat cattatttcg aagttctagt 3001 attctggatc accgtgaagg aagtggacct atagcctttg cccacaagag aatgacacca 3061 tcttccatgc ttaccacagg gaggcaagct caatgtacat cttatttcat tgaacctgta 3121 cagtggatgg aatctgcttt gttgggagtg atggatggac agcttctttg cccaaaatgc 3181 agtgccaagt tgggttcctt caactggtat ggtgaacagt gctcttgtgg taggtggata 3241 acacctgctt ttcaaataca taagaataga gtggatgaaa tgaaaatatt gcctgttttg 3301 ggatcacaaa caggaaaaat aggatccgac tacaaagacc atgacggtga ttataaagat 3361 catgacatcg actacaagga tgacgatgac aagtagcggc cgcgaaggat ctgcgatcgc 3421 tccggtgccc gtcagtgggc agagcgcaca tcgcccacag tccccgagaa gttgggggga 3481 ggggtcggca attgaacggg tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat 3541 gtcgtgtact ggctccgcct ttttcccgag ggtgggggag aaccgtatat aagtgcagta 3601 gtcgccgtga acgttctttt tcgcaacggg tttgccgcca gaacacagct gaagcttcga 3661 ggggctcgca tctctccttc acgcgcccgc cgccctacct gaggccgcca tccacgccgg 3721 ttgagtcgcg ttctgccgcc tcccgcctgt ggtgcctcct gaactgcgtc cgccgtctag 3781 gtaagtttaa agctcaggtc gagaccgggc ctttgtccgg cgctcccttg gagcctacct 3841 agactcagcc ggctctccac gctttgcctg accctgcttg ctcaactcta cgtctttgtt 3901 tcgttttctg ttctgcgccg ttacagatcc aagctgtgac cggcgcctac gctagatgac 3961 cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtcccccggg ccgtacgcac 4021 cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgacc cggaccgcca 4081 catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg 4141 caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt 4201 cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg 4261 gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc 4321 gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc 4381 cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac 4441 ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt 4501 cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgagtcga 4561 caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc 4621 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg 4681 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt 4741 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac 4801 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc 4861 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct 4921 gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct 4981 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct 5041 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct 5101 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggtacctt 5161 taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg 5221 gactggaagg gctaattcac tcccaacgaa aataagatct gctttttgct tgtactgggt 5281 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc 5341 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg 5401 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta 5461 gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga 5521 gtgagaggaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa 5581 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca 5641 atgtatctta tcatgtctgg ctctagctat cccgccccta actccgccca gttccgccca 5701 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc 5761 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctagact tttgcagaga 5821 cggcccaaat tcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac 5881 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt 5941 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc 6001 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg 6061 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt 6121 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa 6181 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc 6241 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag 6301 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt 6361 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg 6421 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg 6481 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg 6541 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac 6601 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg 6661 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt 6721 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg 6781 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc 6841 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt 6901 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt 6961 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag 7021 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt 7081 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc 7141 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc 7201 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg 7261 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac 7321 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg 7381 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc 7441 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact 7501 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc 7561 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat 7621 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc 7681 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac 7741 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa 7801 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact 7861 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg 7921 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg 7981 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag 8041 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca 8101 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc 8161 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc 8221 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag 8281 gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg 8341 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg 8401 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtgc 8461 caagctg //