LOCUS V020899 8947 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V020899 VERSION V020899 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 2293..3801 /label="CYP11B1(NM_000497)" /note="CYP11B1(NM_000497)" /gene="CYP11B1" CDS 3808..3873 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3917..4128 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4141..4409 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 4436..5035 /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 5042..5630 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5704..5937 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6009..6130 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6149..6284 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6322..6338) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6346..6362 /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(6370..6400) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6415..6436 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6724..7312) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7483..8343) /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(8344..8448) /gene="bla" /label="AmpR promoter" primer_bind 8922..8938 /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 tgaccgtcag atcgcctgga gacgccatcc acgctgtttt gacctccata gaagattcta 2281 gagctagcca ccatggcact cagggcaaag gcagaggtgt gcatggcagt gccctggctg 2341 tccctgcaaa gggcacaggc actgggcacg agagccgccc gggtccccag gacagtgctg 2401 ccctttgaag ccatgccccg gcgtccaggc aacaggtggc tgaggctgct gcagatctgg 2461 agggagcagg gttatgagga cctgcacctg gaagtacacc agaccttcca ggaactgggg 2521 cccattttca ggtacgattt gggaggagca ggcatggtgt gtgtgatgct gccggaggac 2581 gtggagaagc tgcaacaggt ggacagcctg catccccaca ggatgagcct ggagccctgg 2641 gtggcctaca gacaacatcg tgggcacaaa tgtggcgtgt tcttgctgaa tgggcctgaa 2701 tggcgcttca accgattgcg gctgaatcca gaagtgctgt cgcccaacgc tgtgcagagg 2761 ttcctcccga tggtggatgc agtggccagg gacttctccc aggccctgaa gaagaaggtg 2821 ctgcagaacg cccgggggag cctgaccctg gacgtccagc ccagcatctt ccactacacc 2881 atagaagcca gcaacttggc tctttttgga gagcggctgg gcctggttgg ccacagcccc 2941 agttctgcca gcctgaactt cctccatgcc ctggaggtca tgttcaaatc caccgtccag 3001 ctcatgttca tgcccaggag cctgtctcgc tggaccagcc ccaaggtgtg gaaggagcac 3061 tttgaggcct gggactgcat cttccagtac ggcgacaact gtatccagaa aatctatcag 3121 gaactggcct tcagccgccc tcaacagtac accagcatcg tggcggagct cctgttgaat 3181 gcggaactgt cgccagatgc catcaaggcc aactctatgg aactcactgc agggagcgtg 3241 gacacgacgg tgtttccctt gctgatgacg ctctttgagc tggctcggaa ccccaacgtg 3301 cagcaggccc tgcgccagga gagcctggcc gccgcagcca gcatcagtga acatccccag 3361 aaggcaacca ccgagctgcc cttgctgcgt gcggccctca aggagacctt gcggctctac 3421 cctgtgggtc tgtttctgga gcgagtggcg agctcagact tggtgcttca gaactaccac 3481 atcccagctg ggacattggt gcgcgtgttc ctctactctc tgggtcgcaa ccccgccttg 3541 ttcccgaggc ctgagcgcta taacccccag cgctggctag acatcagggg ctccggcagg 3601 aacttctacc acgtgccctt tggctttggc atgcgccagt gccttgggcg gcgcctggca 3661 gaggcagaga tgctgctgct gctgcaccat gtgctgaaac acctccaggt ggagacacta 3721 acccaagagg acataaagat ggtctacagc ttcatattga ggcccagcat gttccccctc 3781 ctcaccttca gagccatcaa cggatccgac tacaaagacc atgacggtga ttataaagat 3841 catgacatcg actacaagga tgacgatgac aagtagcggc cgcgaaggat ctgcgatcgc 3901 tccggtgccc gtcagtgggc agagcgcaca tcgcccacag tccccgagaa gttgggggga 3961 ggggtcggca attgaacggg tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat 4021 gtcgtgtact ggctccgcct ttttcccgag ggtgggggag aaccgtatat aagtgcagta 4081 gtcgccgtga acgttctttt tcgcaacggg tttgccgcca gaacacagct gaagcttcga 4141 ggggctcgca tctctccttc acgcgcccgc cgccctacct gaggccgcca tccacgccgg 4201 ttgagtcgcg ttctgccgcc tcccgcctgt ggtgcctcct gaactgcgtc cgccgtctag 4261 gtaagtttaa agctcaggtc gagaccgggc ctttgtccgg cgctcccttg gagcctacct 4321 agactcagcc ggctctccac gctttgcctg accctgcttg ctcaactcta cgtctttgtt 4381 tcgttttctg ttctgcgccg ttacagatcc aagctgtgac cggcgcctac gctagatgac 4441 cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtcccccggg ccgtacgcac 4501 cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgacc cggaccgcca 4561 catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg 4621 caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt 4681 cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg 4741 gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc 4801 gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc 4861 cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac 4921 ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt 4981 cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgagtcga 5041 caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc 5101 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg 5161 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt 5221 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac 5281 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc 5341 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct 5401 gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct 5461 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct 5521 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct 5581 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggtacctt 5641 taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg 5701 gactggaagg gctaattcac tcccaacgaa aataagatct gctttttgct tgtactgggt 5761 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc 5821 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg 5881 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta 5941 gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga 6001 gtgagaggaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa 6061 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca 6121 atgtatctta tcatgtctgg ctctagctat cccgccccta actccgccca gttccgccca 6181 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc 6241 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctagact tttgcagaga 6301 cggcccaaat tcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac 6361 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt 6421 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc 6481 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg 6541 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt 6601 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa 6661 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc 6721 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag 6781 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt 6841 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg 6901 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg 6961 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg 7021 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac 7081 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg 7141 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt 7201 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg 7261 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc 7321 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt 7381 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt 7441 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag 7501 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt 7561 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc 7621 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc 7681 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg 7741 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac 7801 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg 7861 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc 7921 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact 7981 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc 8041 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat 8101 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc 8161 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac 8221 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa 8281 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact 8341 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg 8401 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg 8461 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag 8521 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca 8581 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc 8641 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc 8701 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag 8761 gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg 8821 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg 8881 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtgc 8941 caagctg //