LOCUS V044884 9130 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V044884 VERSION V044884 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" enhancer 1933..2236 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 2237..2440 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 2475..4349 /label="KLHL24(NM_001349413)" /note="KLHL24(NM_001349413)" /gene="KLHL24" CDS 4356..4421 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" CDS 4431..4487 /codon_start=1 /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /product="2A peptide from porcine teschovirus-1 polyprotein" /transl_table=1 /translation="ATNFSLLKQAGDVEENPGP" /label="P2A" CDS 4500..5219 /codon_start=1 /note="mammalian codon-optimized" /product="the original enhanced GFP (Yang et al., 1996)" /transl_table=1 /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL TLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV LLEFVTAAGITLGMDELYK*" /label="EGFP" misc_feature 5226..5813 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5887..6120 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6192..6313 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6332..6467 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6505..6521) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6529..6545 /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(6553..6583) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6598..6619 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6907..7495) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7666..8526) /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(8527..8631) /gene="bla" /label="AmpR promoter" primer_bind 9105..9121 /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 gtcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga 1981 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt 2041 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt 2101 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca 2161 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt 2221 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt 2281 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca 2341 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg 2401 cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat 2461 ccgctagcgc caccatggta ctaatattgg gacgcagact aaacagagag gatcttgggg 2521 tgcgtgattc cccagcaact aagcgaaaag tttttgaaat ggaccccaaa tctctgacag 2581 gtcatgagtt ttttgacttc tcttcaggat catcccatgc cgaaaacata ctccagatat 2641 ttaatgaatt tcgtgatagc cgcttattca cagatgttat catttgtgtg gaaggaaaag 2701 aatttccttg ccatagagct gtgctctcag cctgtagcag ctacttcaga gctatgtttt 2761 gtaatgacca cagggaaagc cgagaaatgt tggttgagat caatggtatt ttagctgaag 2821 ctatggaatg ttttttgcag tatgtttata ctggaaaggt gaagatcact acagagaatg 2881 tacagtatct ctttgagaca tcaagcctct ttcagattag tgttctccgt gatgcatgtg 2941 ccaagttctt ggaggagcaa cttgatcctt gtaattgctt aggaatccag cgctttgctg 3001 atacccattc actcaaaaca ctcttcacaa aatgcaaaaa ttttgcgtta cagacttttg 3061 aggatgtatc ccagcacgaa gaatttcttg agcttgacaa agatgaactt attgattata 3121 tttgtagtga tgaacttgtt attggtaaag aggagatggt ttttgaagcc gtcatgcgtt 3181 gggtctatcg tgccgttgat ctgagaagac cactgttaca cgagctcctg acacatgtga 3241 gactccctct gttgcatccc aactactttg ttcaaacagt tgaagtggac caattgatcc 3301 agaattctcc tgagtgttat cagttgttgc atgaagcaag acggtaccac atacttggga 3361 atgaaatgat gtccccaagg actaggccac gcaggtccac tggctattct gaggtgatag 3421 ttgtcgttgg aggatgtgag cgagttggag gatttaatct tccatacact gagtgctacg 3481 atcctgtaac aggagaatgg aagtctttgg ctaagcttcc agaatttacc aaatcagagt 3541 atgcagtctg tgctctaagg aatgacattc ttgtttcagg tggaagaatc aacagccgtg 3601 atgtctggat ttataactca cagttaaata tttggatcag agttgcctct ctcaataaag 3661 gcagatggcg tcacaaaatg gctgtcctcc ttggtaaagg gtttcttaaa acttctatac 3721 taggccagtc gtggtggctc acacctgtaa tcccagcact ttgggaggcc aaggtatatg 3781 ttgtcggtgg ctatgatggg caaaacagac ttagcagcgt agaatgttat gattcctttt 3841 caaatcgatg gactgaagtt gctcccctta aggaagccgt gagttctcct gcagtgacta 3901 gctgtgtagg caaactgttt gtgattggtg gaggacctga tgataatact tgttctgata 3961 aggttcaatc ttatgatcca gaaaccaatt cttggctact tcgtgcagct atcccaattg 4021 ccaaaaggtg tataacagct gtatccctaa acaacctgat ctatgttgcc ggtggactga 4081 ccaaggcaat atactgttac gatccagttg aagattactg gatgcacgta cagaatacat 4141 tcagccgtca ggaaaactgt ggtatgtctg tgtgtaatgg taaaatatat atcctgggcg 4201 gaagacggga aaatggagaa gccacagaca ctattctctg ttatgatcct gcaacaagta 4261 tcatcacagg ggtagctgca atgcccaggc cagtgtccta tcatggctgt gtgactattc 4321 atagatacaa tgagaaatgc tttaaactcg tcgacgacta caaagaccat gacggtgatt 4381 ataaagatca tgacatcgac tacaaggatg acgatgacaa gggaagcgga gccactaact 4441 tctccctgtt gaaacaagca ggggatgtcg aagagaatcc cgggccagaa ttcgccacca 4501 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg 4561 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg 4621 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc 4681 tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc 4741 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct 4801 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg 4861 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca 4921 agctggagta caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg 4981 gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg 5041 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact 5101 acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc 5161 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaag 5221 gatccaatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg 5281 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt 5341 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg 5401 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc 5461 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc 5521 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc 5581 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc 5641 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg 5701 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc 5761 gtctccgcct tcgccctcag acgagtcgga tctccctttg gccgcctccc cgcctggtac 5821 ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg 5881 ggggactgga agggctaatt cactcccaac gaaaataaga tctgcttttt gcttgtactg 5941 ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac 6001 tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt 6061 gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca 6121 gtagtagttc atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag 6181 agagtgagag gaacttgttt attgcagctt ataatggtta caaataaagc aatagcatca 6241 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca 6301 tcaatgtatc ttatcatgtc tggctctagc tatcccgccc ctaactccgc ccagttccgc 6361 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc 6421 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag acttttgcag 6481 agacggccca aattcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct 6541 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg 6601 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct 6661 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg 6721 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc 6781 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg 6841 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct 6901 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca 6961 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct 7021 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc 7081 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt 7141 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc 7201 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc 7261 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg 7321 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc 7381 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag 7441 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga 7501 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat 7561 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag 7621 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat 7681 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc 7741 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat 7801 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag 7861 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg 7921 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc 7981 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca 8041 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg 8101 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc 8161 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta 8221 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc 8281 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg 8341 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc 8401 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc 8461 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat 8521 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag 8581 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc 8641 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa 8701 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg 8761 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca 8821 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc 8881 atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt 8941 aaggagaaaa taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg 9001 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag 9061 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag 9121 tgccaagctg //