LOCUS V035970 8707 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V035970 VERSION V035970 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..3561 /label="Cpa4(NM_027926)" /note="Cpa4(NM_027926)" /gene="Cpa4" CDS 3568..3633 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3677..3888 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3901..4169 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 4196..4795 /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 4802..5390 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5464..5697 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 5769..5890 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 5909..6044 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6082..6098) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6106..6122 /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(6130..6160) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6175..6196 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6484..7072) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7243..8103) /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(8104..8208) /gene="bla" /label="AmpR promoter" primer_bind 8682..8698 /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 catgaagtgg ctactgttct ttggggccct gattggggca 2341 ggcatctgcg gccgagataa attctttggg gaccaagttt ttaggattaa tgtcagaaat 2401 ggagacgaga tcagaaaact gactgagcta gtgaattctg accacttaaa gctcagtgtc 2461 tggaaatctc cctccacttt tgatcggccc gtggatattc tcgtcccttc tgtcagcttg 2521 ctcccagtca agtccttcct gaagtcccag ggtttagact actccgtgac aattgaggac 2581 ctccaggccc ttttagataa cgaagatgaa gaaatgcagc acaatgaagg gatagagcgg 2641 agtggtgact tcaactacgg agcttaccat cccctggagg ctatttacca cgagatggac 2701 agtattgcca cagactttcc tgagctggtg agcagagtga agattggaga gacatttgaa 2761 aagcggccca tgtatgttct aaagttcagc acgggaggag gcaagaagcg gccagccatt 2821 tggttgaatg caggcatcca tgcccgtgag tggatctcac aggccacagc catctggaca 2881 gcgaggaaga ttgtaactga ttataaaaag gacccggcta tcacctccat cttgaagaaa 2941 gtggatattt tcttgttgcc cgtggccaat cctgatggat atgtgtacac gcaaagccag 3001 aaccgattat ggagaaagac gcggtcccgg aacccaggaa gccgctgtgt tggagccgat 3061 ccaaacagaa actggaatgc tagttttgca ggagagggga ccagtgataa cccttgctct 3121 gaagtgtacc atggctccca ccccaattct gaagtggagg tgaaatcggt ggtggatttc 3181 attcaaaagc atgggaactt caaatgcttc attgacctgc atagctactc acagctgctg 3241 atgtatccct atgggtacac agtcaagaag gccccggatg ctgaggagct ggacgatgtg 3301 gcgaggaatg cagcccaagc cctggcttct ctctcgggca ctaagtaccg agtgggccca 3361 acttgcacca ccgtctatcc agctagtggg agcagcgttg actgggcata cgacaatggc 3421 atcaagtatg ctttcacatt tgagctgaga gacactgggt actacggctt cctcttgcca 3481 gccagccaga tcatccccac cgcggaggag acctggctag ggctgaagac catcatggaa 3541 catgttcggg accacctcta cggatccgac tacaaagacc atgacggtga ttataaagat 3601 catgacatcg actacaagga tgacgatgac aagtagcggc cgcgaaggat ctgcgatcgc 3661 tccggtgccc gtcagtgggc agagcgcaca tcgcccacag tccccgagaa gttgggggga 3721 ggggtcggca attgaacggg tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat 3781 gtcgtgtact ggctccgcct ttttcccgag ggtgggggag aaccgtatat aagtgcagta 3841 gtcgccgtga acgttctttt tcgcaacggg tttgccgcca gaacacagct gaagcttcga 3901 ggggctcgca tctctccttc acgcgcccgc cgccctacct gaggccgcca tccacgccgg 3961 ttgagtcgcg ttctgccgcc tcccgcctgt ggtgcctcct gaactgcgtc cgccgtctag 4021 gtaagtttaa agctcaggtc gagaccgggc ctttgtccgg cgctcccttg gagcctacct 4081 agactcagcc ggctctccac gctttgcctg accctgcttg ctcaactcta cgtctttgtt 4141 tcgttttctg ttctgcgccg ttacagatcc aagctgtgac cggcgcctac gctagatgac 4201 cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtcccccggg ccgtacgcac 4261 cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgacc cggaccgcca 4321 catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg 4381 caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt 4441 cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg 4501 gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc 4561 gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc 4621 cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac 4681 ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt 4741 cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgagtcga 4801 caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc 4861 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg 4921 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt 4981 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac 5041 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc 5101 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct 5161 gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct 5221 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct 5281 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct 5341 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggtacctt 5401 taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg 5461 gactggaagg gctaattcac tcccaacgaa aataagatct gctttttgct tgtactgggt 5521 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc 5581 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg 5641 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta 5701 gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga 5761 gtgagaggaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa 5821 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca 5881 atgtatctta tcatgtctgg ctctagctat cccgccccta actccgccca gttccgccca 5941 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc 6001 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctagact tttgcagaga 6061 cggcccaaat tcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac 6121 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt 6181 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc 6241 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg 6301 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt 6361 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa 6421 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc 6481 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag 6541 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt 6601 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg 6661 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg 6721 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg 6781 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac 6841 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg 6901 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt 6961 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg 7021 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc 7081 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt 7141 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt 7201 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag 7261 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt 7321 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc 7381 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc 7441 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg 7501 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac 7561 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg 7621 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc 7681 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact 7741 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc 7801 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat 7861 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc 7921 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac 7981 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa 8041 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact 8101 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg 8161 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg 8221 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag 8281 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca 8341 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc 8401 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc 8461 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag 8521 gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg 8581 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg 8641 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtgc 8701 caagctg //