LOCUS V026012 7924 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V026012 VERSION V026012 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..2973 /label="CHMP4B(NM_176812)" /note="CHMP4B(NM_176812)" /gene="CHMP4B" CDS 2980..3045 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3089..3300 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3313..3581 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 3614..4012 /codon_start=1 /gene="Aspergillus terreus BSD" /note="confers resistance to blasticidin" /product="blasticidin S deaminase" /transl_table=1 /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI KAIVKDSDGQPTAVGIRELLPSGYVWEG*" /label="BSD" misc_feature 4019..4607 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 4681..4914 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 4986..5107 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 5126..5261 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(5299..5315) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5323..5339 /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(5347..5377) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5392..5413 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5701..6289) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6460..7320) /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(7321..7425) /gene="bla" /label="AmpR promoter" primer_bind 7899..7915 /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 catgtcggtg ttcgggaagc tgttcggggc tggagggggt 2341 aaggccggca agggcggccc gaccccccag gaggccatcc agcggctgcg ggacacggaa 2401 gagatgttaa gcaagaaaca ggagttcctg gagaagaaaa tcgagcagga gctgacggcc 2461 gccaagaagc acggcaccaa aaacaagcgc gcggccctcc aggcactgaa gcgtaagaag 2521 aggtatgaga agcagctggc gcagatcgac ggcacattat caaccatcga gttccagcgg 2581 gaggccctgg agaatgccaa caccaacacc gaggtgctca agaacatggg ctatgccgcc 2641 aaggccatga aggcggccca tgacaacatg gacatcgata aagttgatga gttaatgcag 2701 gacattgctg accagcaaga acttgcagag gagatttcaa cagcaatttc gaaacctgta 2761 gggtttggag aagagtttga cgaggatgag ctcatggcgg aattagaaga actagaacag 2821 gaggaactag acaagaattt gctggaaatc agtggacccg aaacagtccc tctaccaaat 2881 gttccctcta tagccctacc atcaaaaccc gccaagaaga aagaagagga ggacgacgac 2941 atgaaggaat tggagaactg ggctggatcc atgggatccg actacaaaga ccatgacggt 3001 gattataaag atcatgacat cgactacaag gatgacgatg acaagtagcg gccgcgaagg 3061 atctgcgatc gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag 3121 aagttggggg gaggggtcgg caattgaacg ggtgcctaga gaaggtggcg cggggtaaac 3181 tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat 3241 ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag 3301 ctgaagcttc gaggggctcg catctctcct tcacgcgccc gccgccctac ctgaggccgc 3361 catccacgcc ggttgagtcg cgttctgccg cctcccgcct gtggtgcctc ctgaactgcg 3421 tccgccgtct aggtaagttt aaagctcagg tcgagaccgg gcctttgtcc ggcgctccct 3481 tggagcctac ctagactcag ccggctctcc acgctttgcc tgaccctgct tgctcaactc 3541 tacgtctttg tttcgttttc tgttctgcgc cgttacagat ccaagctgtg accggcgcct 3601 acgctaggcc accatggcca agcctttgtc tcaagaagaa tccaccctca ttgaaagagc 3661 aacggctaca atcaacagca tccccatctc tgaagactac agcgtcgcca gcgcagctct 3721 ctctagcgac ggccgcatct tcactggtgt caatgtatat cattttactg ggggaccttg 3781 tgcagaactc gtggtgctgg gcactgctgc tgctgcggca gctggcaacc tgacttgtat 3841 cgtcgcgatc ggaaatgaga acaggggcat cttgagcccc tgcggacggt gccgacaggt 3901 gcttctcgat ctgcatcctg ggatcaaagc catagtgaag gacagtgatg gacagccgac 3961 ggcagttggg attcgtgaat tgctgccctc tggttatgtg tgggagggct aagtcgacaa 4021 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc 4081 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat 4141 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg 4201 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg 4261 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat 4321 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt 4381 gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc 4441 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa 4501 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg 4561 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg gtacctttaa 4621 gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac 4681 tggaagggct aattcactcc caacgaaaat aagatctgct ttttgcttgt actgggtctc 4741 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta 4801 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact 4861 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtagta 4921 gttcatgtca tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg 4981 agaggaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt 5041 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg 5101 tatcttatca tgtctggctc tagctatccc gcccctaact ccgcccagtt ccgcccattc 5161 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc 5221 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctagactttt gcagagacgg 5281 cccaaattcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat 5341 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag 5401 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg 5461 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc 5521 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc 5581 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa 5641 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt 5701 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg 5761 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg 5821 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag 5881 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc 5941 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa 6001 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg 6061 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc 6121 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac 6181 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg 6241 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt 6301 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt 6361 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa 6421 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga 6481 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt 6541 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg 6601 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga 6661 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga 6721 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg 6781 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc 6841 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc 6901 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca 6961 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac 7021 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg 7081 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc 7141 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg 7201 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac 7261 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat 7321 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata 7381 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa 7441 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg 7501 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat 7561 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg 7621 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga 7681 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag 7741 aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc 7801 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt 7861 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa 7921 gctg //