LOCUS V035217 9571 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V035217 VERSION V035217 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..4425 /label="FBXO43(NM_001029860)" /note="FBXO43(NM_001029860)" /gene="FBXO43" CDS 4432..4497 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 4541..4752 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4765..5033 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 5060..5659 /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 5666..6254 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6328..6561 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6633..6754 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6773..6908 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6946..6962) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6970..6986 /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(6994..7024) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7039..7060 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7348..7936) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8107..8967) /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(8968..9072) /gene="bla" /label="AmpR promoter" primer_bind 9546..9562 /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 catgagtttt aaagacaaag atgagagaat ttcttgtttg 2341 gaagcctacg taactttgac atctaagagc tcaagattta ctgatgaaac agagattttg 2401 aagatgtcgc aaaggcactc aggtcaagct ggcactgaag caggaaatgg ggcggactct 2461 cctccaattg tcaactccaa gtactccacc ttcagagatt tttgttccac atcttcattt 2521 caagatagtg gctacaatga gttaaaatct tgtagctttg ataatataga taaagaatat 2581 cttggaaaga aagaaaaagg cccaacatta ctctatgagc accctgaaac ttcaggcctg 2641 ggcttaacac atcctttaga atctcccact caaaaaaaga aatgtatctt gcctagaaag 2701 gaaaaggata aaaccccaga actttgtgaa acacctaaaa tcagtgggaa aaaatgttta 2761 cctcgcagaa ggttgaatgt atctttcgct cttctaaaag gggactttga atcacaaaat 2821 agttctttag aaagtagtat aagccaagtt atcaacttag aaaaaaatat tccaagcagt 2881 gcttcaggtt tttccagggc aaataatttt agccctttag ttactagcac tttaaaaaca 2941 gaagaagtga cttcatgcag tcaaaaattg aggcttaatt tttctcagca aaagacttcc 3001 acaattgatg attccaaaga tgattgtagc ctatttgaag ttgaatgtat atctccaatt 3061 cagggcaata attttaaaga ctctatcaca catgacttta gtgatagcag tttatgcatt 3121 aatgatgaga atgcatgtcc agagctcctg ggctcctctg ttagtggaac aacttgtgga 3181 acagatgagg acatatttgt gactccgata agtaatcttg tggcaaacat tagatttaac 3241 gcaagtcaaa tactttctcc ttcacctgaa gtgagaggca gtatttcaac gcctgaagac 3301 agtggtttta actcacttag cttggagaaa tcagaagatt ccctgtctga ccaggagggt 3361 tcttttcaag aactactgca gaaacataag gggactccca aagttgggga caccataaga 3421 aagacaagac atcttggaag gtcgagaaga ctgtccaccc ttcgggaaca aagctcgcag 3481 tcagagacag aagaggaaaa gcagattgtc caccctgact ctgaaaaaag agcagcagct 3541 gcttctgcca tctcagaggg tcagctgagt agtgatgaga gtggggattt gacctttagc 3601 ttaaagaatt tatcaaagac cccagccttg caattggtac atgagctgtt catgaaaagc 3661 aagaggaaaa gattacagga aaatagtgga catgaattct tagagcaagg ggatggggag 3721 aaaatagctg tactgcagtg tatacttgca ggactgatcg gcaagaaaat gggtatagaa 3781 aaactggaca tcttaacaga attaaaatat agaaatttaa agcatattct tgctatggtt 3841 ttagagtcct tgaccgcaga gagcctatgc agtgtttgga aagtaagcag aaattggcgt 3901 gaaattgttg ttcaagataa aaatgcaaat cggaggagga aattttatat cacacaactg 3961 aaaacagatt ctgagggggc tgtattaaat gtcgaggatg ctgccactcg gctccagctt 4021 ttaaatcgct cagctttaag atctgtgcag gcacaggcta ggatacctgg ttctcagaga 4081 gagcaagggt caacattatc tccctgggga gaagttttga cacctctagc aagctcttct 4141 gttactcact taagtagtaa acaggaagaa tatgttaagg ttgccaaaac actttttact 4201 gatgaagcat taaaaccttg cccaaggtgc cagtcccctg ctaagtacca gccatataag 4261 aaaaggggac tgtgtagccg aacagcctgt ggttttgact tttgtgtgtt atgtctgtgt 4321 gcttatcatg ggtctgaaga atgtagtaga ggagcagcaa agccaagaaa tagaaaagat 4381 gctctcccag gaagtgccca gagtaagcgg aatttaaaac gcctcggatc cgactacaaa 4441 gaccatgacg gtgattataa agatcatgac atcgactaca aggatgacga tgacaagtag 4501 cggccgcgaa ggatctgcga tcgctccggt gcccgtcagt gggcagagcg cacatcgccc 4561 acagtccccg agaagttggg gggaggggtc ggcaattgaa cgggtgccta gagaaggtgg 4621 cgcggggtaa actgggaaag tgatgtcgtg tactggctcc gcctttttcc cgagggtggg 4681 ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc 4741 gccagaacac agctgaagct tcgaggggct cgcatctctc cttcacgcgc ccgccgccct 4801 acctgaggcc gccatccacg ccggttgagt cgcgttctgc cgcctcccgc ctgtggtgcc 4861 tcctgaactg cgtccgccgt ctaggtaagt ttaaagctca ggtcgagacc gggcctttgt 4921 ccggcgctcc cttggagcct acctagactc agccggctct ccacgctttg cctgaccctg 4981 cttgctcaac tctacgtctt tgtttcgttt tctgttctgc gccgttacag atccaagctg 5041 tgaccggcgc ctacgctaga tgaccgagta caagcccacg gtgcgcctcg ccacccgcga 5101 cgacgtcccc cgggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg 5161 ccacaccgtc gacccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct 5221 cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc 5281 ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg 5341 catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc 5401 gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca 5461 ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc 5521 cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct 5581 cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac 5641 ccgcaagccc ggtgcctgag tcgacaatca acctctggat tacaaaattt gtgaaagatt 5701 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc 5761 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg 5821 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac 5881 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc 5941 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc 6001 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa 6061 atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc 6121 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc 6181 ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg 6241 ggccgcctcc ccgcctggta cctttaagac caatgactta caaggcagct gtagatctta 6301 gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa cgaaaataag 6361 atctgctttt tgcttgtact gggtctctct ggttagacca gatctgagcc tgggagctct 6421 ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag 6481 tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt 6541 cagtgtggaa aatctctagc agtagtagtt catgtcatct tattattcag tatttataac 6601 ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt tattgcagct tataatggtt 6661 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta 6721 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggctctag ctatcccgcc 6781 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta 6841 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt 6901 tggaggccta gacttttgca gagacggccc aaattcgtaa tcatggtcat agctgtttcc 6961 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg 7021 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc 7081 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg 7141 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc 7201 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac 7261 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa 7321 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca 7381 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc 7441 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata 7501 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta 7561 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca 7621 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga 7681 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg 7741 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg 7801 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg 7861 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag 7921 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa 7981 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat 8041 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc 8101 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc 8161 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc 8221 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc 8281 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc 8341 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt 8401 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc 8461 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa 8521 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt 8581 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg 8641 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc 8701 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa 8761 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt 8821 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt 8881 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag 8941 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta 9001 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat 9061 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat 9121 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg 9181 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta 9241 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg 9301 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg 9361 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag 9421 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc 9481 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg 9541 acgttgtaaa acgacggcca gtgccaagct g //