LOCUS V030472 9844 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V030472 VERSION V030472 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..4695 /label="USP10(NM_005153)" /note="USP10(NM_005153)" /gene="USP10" CDS 4705..4770 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 4814..5025 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 5038..5306 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 5333..5932 /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 5939..6527 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6601..6834 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6906..7027 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 7046..7181 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(7219..7235) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7243..7259 /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(7267..7297) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7312..7333 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7621..8209) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8380..9240) /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(9241..9345) /gene="bla" /label="AmpR promoter" primer_bind 9819..9835 /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 catggccctc cacagcccgc agtatatttt tggagatttt 2341 agccctgatg aattcaatca attctttgtg actcctcgat cttcagttga gcttcctcca 2401 tacagtggaa cagttctgtg tggcacacag gctgtggata aactacctga tggacaagaa 2461 tatcagagaa ttgagtttgg tgtcgatgaa gtcattgaac ccagtgacac tttgccgaga 2521 acccccagct acagtatttc aagcacactg aaccctcagg cccctgaatt tattctcggt 2581 tgtacagctt ccaaaataac ccctgatggt atcactaaag aagcaagcta tggctccatc 2641 gactgccagt acccaggctc tgccctcgct ttggatggaa gttctaatgt ggaggcggaa 2701 gttttggaaa atgatggtgt ctcaggtggt cttggacaaa gggagcgtaa aaagaagaaa 2761 aagcggccac ctggatatta cagctatttg aaagatggtg gcgatgatag tatctccaca 2821 gaagccctgg tcaatggcca tgccaattca gcagtcccga acagtgtcag tgcagaggat 2881 gcagaattta tgggtgacat gcccccgtca gttacgccca ggacttgtaa cagcccccag 2941 aactccacag actctgtcag tgacattgtg cctgacagtc ctttccccgg agcactcggc 3001 agtgacacca ggactgcagg gcagccagag gggggccccg gggctgattt tggtcagtcc 3061 tgcttccctg cagaggctgg cagagacacc ctgtcaagga cagctggggc tcagccctgc 3121 gttggtaccg atactactga aaaccttgga gttgctaatg gacaaatact tgaatcctcg 3181 ggtgagggca cagctaccaa cggggtggag ttgcacacca cggaaagcat agacttggac 3241 ccaaccaaac ccgagagtgc atcacctcct gctgacggca cgggctctgc atcaggcacc 3301 cttcctgtca gccagcccaa gtcctgggcc agcctctttc atgattctaa gccctcttcc 3361 tcctcgccgg tggcctatgt ggaaactaag tattcccctc ccgccatatc tcccctggtt 3421 tctgaaaagc aggttgaagt caaagaaggg cttgttccgg tttcagagga tcctgtagcc 3481 ataaagattg cagagttgct ggagaatgta accctaatcc ataaaccagt gtcgttgcaa 3541 ccccgtgggc tgatcaataa agggaactgg tgctacatta atgctacact gcaggcattg 3601 gttgcttgcc cgccgatgta ccacctgatg aagttcattc ctctgtattc caaagtgcaa 3661 aggccttgta cgtcaacacc catgatagac agctttgttc ggctaatgaa tgagttcact 3721 aatatgccag tacctccaaa accccgacaa gctcttggag ataaaatcgt gagggatatt 3781 cgccctggag ctgcctttga gcccacatat atttacagac tcctgacagt taacaagtca 3841 agcctgtctg aaaagggtcg acaagaagat gctgaggaat acttaggctt cattctaaat 3901 ggacttcatg aggaaatgtt gaacctaaag aagcttctct caccaagtaa tgaaaaactt 3961 acgatttcca acggccccaa aaaccactcg gtcaatgaag aagagcagga agaacaaggt 4021 gaaggaagcg aggatgaatg ggaacaagtg ggcccccgga acaagacttc cgtcacccgc 4081 caggcggatt ttgttcagac tccaatcacc ggcatttttg gtggacacat caggtctgtg 4141 gtttaccagc agagttcaaa agaatctgcc actttgcagc catttttcac gttgcagttg 4201 gatatccagt cagacaagat acgcacagtc caggatgcac tggagagctt ggtggcaaga 4261 gaatctgtcc aaggttatac cacaaaaacc aaacaagagg ttgagataag tcgaagagtg 4321 actctggaaa aactccctcc tgtcctcgtg ctgcacctga aacgattcgt ttatgagaag 4381 actggtgggt gccagaagct tatcaaaaat attgaatatc ctgtggactt ggaaattagt 4441 aaagaactgc tttctccagg ggttaaaaat aagaatttta aatgccaccg aacctatcgg 4501 ctctttgcag tggtctacca tcacggcaac agtgcgacgg gcggccatta cactacagac 4561 gtcttccaga tcggtctgaa tggctggctg cgcatcgatg accagacagt caaggtgatc 4621 aaccagtacc aggtggtgaa accaactgct gaacgcacag cctacctcct gtattaccgc 4681 cgagtggacc tgctgtaagg atccgactac aaagaccatg acggtgatta taaagatcat 4741 gacatcgact acaaggatga cgatgacaag tagcggccgc gaaggatctg cgatcgctcc 4801 ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg 4861 gtcggcaatt gaacgggtgc ctagagaagg tggcgcgggg taaactggga aagtgatgtc 4921 gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag tgcagtagtc 4981 gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacagctgaa gcttcgaggg 5041 gctcgcatct ctccttcacg cgcccgccgc cctacctgag gccgccatcc acgccggttg 5101 agtcgcgttc tgccgcctcc cgcctgtggt gcctcctgaa ctgcgtccgc cgtctaggta 5161 agtttaaagc tcaggtcgag accgggcctt tgtccggcgc tcccttggag cctacctaga 5221 ctcagccggc tctccacgct ttgcctgacc ctgcttgctc aactctacgt ctttgtttcg 5281 ttttctgttc tgcgccgtta cagatccaag ctgtgaccgg cgcctacgct agatgaccga 5341 gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc ccccgggccg tacgcaccct 5401 cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgacccgg accgccacat 5461 cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa 5521 ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga 5581 agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct 5641 ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg 5701 gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt 5761 cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc 5821 cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga 5881 ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gagtcgacaa 5941 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc 6001 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat 6061 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg 6121 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg 6181 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat 6241 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt 6301 gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc 6361 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa 6421 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg 6481 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg gtacctttaa 6541 gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac 6601 tggaagggct aattcactcc caacgaaaat aagatctgct ttttgcttgt actgggtctc 6661 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta 6721 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact 6781 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtagta 6841 gttcatgtca tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg 6901 agaggaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt 6961 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg 7021 tatcttatca tgtctggctc tagctatccc gcccctaact ccgcccagtt ccgcccattc 7081 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc 7141 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctagactttt gcagagacgg 7201 cccaaattcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat 7261 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag 7321 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg 7381 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc 7441 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc 7501 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa 7561 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt 7621 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg 7681 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg 7741 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag 7801 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc 7861 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa 7921 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg 7981 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc 8041 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac 8101 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg 8161 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt 8221 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt 8281 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa 8341 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga 8401 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt 8461 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg 8521 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga 8581 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga 8641 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg 8701 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc 8761 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc 8821 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca 8881 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac 8941 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg 9001 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc 9061 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg 9121 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac 9181 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat 9241 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata 9301 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa 9361 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg 9421 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat 9481 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg 9541 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga 9601 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag 9661 aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc 9721 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt 9781 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa 9841 gctg //