LOCUS V035300 8227 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V035300 VERSION V035300 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..3081 /label="KLK8(NM_007196)" /note="KLK8(NM_007196)" /gene="KLK8" CDS 3088..3153 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3197..3408 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3421..3689 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 3716..4315 /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 4322..4910 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 4984..5217 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 5289..5410 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 5429..5564 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(5602..5618) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5626..5642 /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(5650..5680) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5695..5716 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6004..6592) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6763..7623) /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(7624..7728) /gene="bla" /label="AmpR promoter" primer_bind 8202..8218 /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 catgggacgc ccccgacctc gtgcggccaa gacgtggatg 2341 ttcctgctct tgctgggggg agcctgggca ggacactcca gggcacagga ggacaaggtg 2401 ctggggggtc atgagtgcca accccattcg cagccttggc aggcggcctt gttccagggc 2461 cagcaactac tctgtggcgg tgtccttgta ggtggcaact gggtccttac agctgcccac 2521 tgtaaaaaac cgaaatacac agtacgcctg ggagaccaca gcctacagaa taaagatggc 2581 ccagagcaag aaatacctgt ggttcagtcc atcccacacc cctgctacaa cagcagcgat 2641 gtggaggacc acaaccatga tctgatgctt cttcaactgc gtgaccaggc atccctgggg 2701 tccaaagtga agcccatcag cctggcagat cattgcaccc agcctggcca gaagtgcacc 2761 gtctcaggct ggggcactgt caccagtccc cgagagaatt ttcctgacac tctcaactgt 2821 gcagaagtaa aaatctttcc ccagaagaag tgtgaggatg cttacccggg gcagatcaca 2881 gatggcatgg tctgtgcagg cagcagcaaa ggggctgaca cgtgccaggg cgattctgga 2941 ggccccctgg tgtgtgatgg tgcactccag ggcatcacat cctggggctc agacccctgt 3001 gggaggtccg acaaacctgg cgtctatacc aacatctgcc gctacctgga ctggatcaag 3061 aagatcatag gcagcaaggg cggatccgac tacaaagacc atgacggtga ttataaagat 3121 catgacatcg actacaagga tgacgatgac aagtagcggc cgcgaaggat ctgcgatcgc 3181 tccggtgccc gtcagtgggc agagcgcaca tcgcccacag tccccgagaa gttgggggga 3241 ggggtcggca attgaacggg tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat 3301 gtcgtgtact ggctccgcct ttttcccgag ggtgggggag aaccgtatat aagtgcagta 3361 gtcgccgtga acgttctttt tcgcaacggg tttgccgcca gaacacagct gaagcttcga 3421 ggggctcgca tctctccttc acgcgcccgc cgccctacct gaggccgcca tccacgccgg 3481 ttgagtcgcg ttctgccgcc tcccgcctgt ggtgcctcct gaactgcgtc cgccgtctag 3541 gtaagtttaa agctcaggtc gagaccgggc ctttgtccgg cgctcccttg gagcctacct 3601 agactcagcc ggctctccac gctttgcctg accctgcttg ctcaactcta cgtctttgtt 3661 tcgttttctg ttctgcgccg ttacagatcc aagctgtgac cggcgcctac gctagatgac 3721 cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtcccccggg ccgtacgcac 3781 cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgacc cggaccgcca 3841 catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg 3901 caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt 3961 cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg 4021 gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc 4081 gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc 4141 cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac 4201 ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt 4261 cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgagtcga 4321 caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc 4381 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg 4441 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt 4501 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac 4561 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc 4621 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct 4681 gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct 4741 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct 4801 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct 4861 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggtacctt 4921 taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg 4981 gactggaagg gctaattcac tcccaacgaa aataagatct gctttttgct tgtactgggt 5041 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc 5101 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg 5161 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta 5221 gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga 5281 gtgagaggaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa 5341 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca 5401 atgtatctta tcatgtctgg ctctagctat cccgccccta actccgccca gttccgccca 5461 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc 5521 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctagact tttgcagaga 5581 cggcccaaat tcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac 5641 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt 5701 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc 5761 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg 5821 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt 5881 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa 5941 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc 6001 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag 6061 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt 6121 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg 6181 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg 6241 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg 6301 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac 6361 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg 6421 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt 6481 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg 6541 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc 6601 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt 6661 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt 6721 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag 6781 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt 6841 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc 6901 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc 6961 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg 7021 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac 7081 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg 7141 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc 7201 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact 7261 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc 7321 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat 7381 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc 7441 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac 7501 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa 7561 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact 7621 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg 7681 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg 7741 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag 7801 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca 7861 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc 7921 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc 7981 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag 8041 gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg 8101 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg 8161 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtgc 8221 caagctg //