LOCUS V022685 8884 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V022685 VERSION V022685 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..3732 /label="OSGIN1(NM_182981)" /note="OSGIN1(NM_182981)" /gene="OSGIN1" CDS 3739..3804 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3854..4065 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4078..4346 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 4373..4972 /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 4979..5567 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5641..5874 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 5946..6067 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6086..6221 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6259..6275) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6283..6299 /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(6307..6337) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6352..6373 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6661..7249) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7420..8280) /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(8281..8385) /gene="bla" /label="AmpR promoter" primer_bind 8859..8875 /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 catgagctcc tccagaaagg accacctcgg cgccagcagc 2341 tcagagcccc tcccggtcat cattgtgggt aacggcccct ctggtatctg cctgtcctac 2401 ctgctctccg gctacacacc ctacacgaag ccagatgcca tccacccaca ccccctgctg 2461 cagaggaagc tcaccgaggc cccgggggtc tccatcctgg accaggacct ggactacctg 2521 tccgaaggcc tcgaaggccg atcccaaagc cccgtggccc tgctctttga tgcccttcta 2581 cgcccagaca cagactttgg gggaaacatg aagtcggtcc tcacctggaa gcaccggaag 2641 gagcacgcca tcccccacgt ggttctgggc cggaacctcc ccgggggagc ctggcactcc 2701 atcgaaggct ccatggtgat cctgagccaa ggccagtgga tggggctccc ggacctggag 2761 gtcaaggact ggatgcagaa gaagcgaaga ggtcttcgca acagccgggc cactgccggg 2821 gacatcgccc actactacag ggactacgtg gtcaagaagg gtctggggca taactttgtg 2881 tccggtgctg tagtcacagc cgtggagtgg gggacccccg atcccagcag ctgtggggcc 2941 caggactcca gccccctctt ccaggtgagc ggcttcctga ccaggaacca ggcccagcag 3001 cccttctcgc tgtgggcccg caacgtggtc ctcgccacag gcacgttcga cagcccggcc 3061 cggctgggca tccccgggga ggccctgccc ttcatccacc atgagctgtc tgccctggag 3121 gccgccacaa gggtgggtgc ggtgaccccg gcctcagacc ctgtcctcat cattggcgcg 3181 gggctgtcag cggccgacgc ggtcctctac gcccgccact acaacatccc ggtgatccat 3241 gccttccgcc gggccgtgga cgaccctggc ctggtgttca accagctgcc caagatgctg 3301 taccccgagt accacaaggt gcaccagatg atgcgggagc agtccatcct gtcgcccagc 3361 ccctatgagg gttaccgcag cctccccagg caccagctgc tgtgcttcaa ggaagactgc 3421 caggccgtgt tccaggacct cgagggtgtc gagaaggtgt ttggggtctc cctggtgctg 3481 gtcctcatcg gctcccaccc cgacctctcc ttcctgcctg gggcaggggc tgactttgca 3541 gtggatcctg accagccgct gagcgccaag aggaacccca ttgacgtgga ccccttcacc 3601 taccagagca cccgccagga gggcctgtac gccatggggc cgctggccgg ggacaacttc 3661 gtgaggtttg tgcagggggg cgccttggct gtggccagct ccctgctaag gaaggagacc 3721 aggaagccac ccctcgagga ctacaaagac catgacggtg attataaaga tcatgacatc 3781 gactacaagg atgacgatga caagtaagat ccgcggccgc gaaggatctg cgatcgctcc 3841 ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg 3901 gtcggcaatt gaacgggtgc ctagagaagg tggcgcgggg taaactggga aagtgatgtc 3961 gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag tgcagtagtc 4021 gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacagctgaa gcttcgaggg 4081 gctcgcatct ctccttcacg cgcccgccgc cctacctgag gccgccatcc acgccggttg 4141 agtcgcgttc tgccgcctcc cgcctgtggt gcctcctgaa ctgcgtccgc cgtctaggta 4201 agtttaaagc tcaggtcgag accgggcctt tgtccggcgc tcccttggag cctacctaga 4261 ctcagccggc tctccacgct ttgcctgacc ctgcttgctc aactctacgt ctttgtttcg 4321 ttttctgttc tgcgccgtta cagatccaag ctgtgaccgg cgcctacgct agatgaccga 4381 gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc ccccgggccg tacgcaccct 4441 cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgacccgg accgccacat 4501 cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa 4561 ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga 4621 agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct 4681 ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg 4741 gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt 4801 cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc 4861 cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga 4921 ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gagtcgacaa 4981 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc 5041 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat 5101 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg 5161 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg 5221 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat 5281 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt 5341 gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc 5401 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa 5461 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg 5521 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg gtacctttaa 5581 gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac 5641 tggaagggct aattcactcc caacgaaaat aagatctgct ttttgcttgt actgggtctc 5701 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta 5761 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact 5821 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtagta 5881 gttcatgtca tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg 5941 agaggaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt 6001 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg 6061 tatcttatca tgtctggctc tagctatccc gcccctaact ccgcccagtt ccgcccattc 6121 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc 6181 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctagactttt gcagagacgg 6241 cccaaattcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat 6301 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag 6361 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg 6421 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc 6481 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc 6541 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa 6601 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt 6661 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg 6721 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg 6781 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag 6841 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc 6901 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa 6961 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg 7021 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc 7081 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac 7141 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg 7201 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt 7261 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt 7321 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa 7381 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga 7441 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt 7501 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg 7561 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga 7621 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga 7681 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg 7741 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc 7801 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc 7861 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca 7921 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac 7981 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg 8041 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc 8101 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg 8161 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac 8221 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat 8281 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata 8341 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa 8401 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg 8461 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat 8521 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg 8581 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga 8641 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag 8701 aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc 8761 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt 8821 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa 8881 gctg //