LOCUS V030490 9744 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V030490 VERSION V030490 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" regulatory 2288..2297 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 2297..2362 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" CDS 2369..4669 /label="NSUN2(NM_017755)" /note="NSUN2(NM_017755)" /gene="NSUN2" promoter 4714..4925 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4938..5206 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 5233..5832 /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 5839..6427 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6501..6734 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6806..6927 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6946..7081 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(7119..7135) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7143..7159 /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(7167..7197) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7212..7233 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7521..8109) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8280..9140) /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(9141..9245) /gene="bla" /label="AmpR promoter" primer_bind 9719..9735 /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 agagctagcc accatggact acaaagacca tgacggtgat tataaagatc atgacatcga 2341 ctacaaggat gacgatgaca aggaattcat ggggcggcgg tcgcggggtc ggcggctcca 2401 gcaacagcag cggccggagg acgcggagga tggcgccgag ggtggtggaa agcgcggcga 2461 ggcgggctgg gaaggaggct accccgagat cgtcaaggag aacaagctgt tcgagcacta 2521 ctaccaggag ctcaagatcg tgcccgaggg cgagtggggc cagttcatgg acgctctcag 2581 ggagccgctc ccggccactt taagaattac tggttacaaa agccacgcaa aagagattct 2641 ccattgctta aagaacaaat attttaagga attggaggac ctggaggtgg acggtcagaa 2701 agttgaagtt ccacagccac tgagttggta tcctgaagaa cttgcctggc acacaaattt 2761 aagtcgaaaa atcttgagaa aatcgccaca cttggaaaag tttcatcagt ttctagttag 2821 tgaaacagaa tctggaaata ttagtcgtca agaagctgtt agcatgatcc caccactgct 2881 cctcaacgtg cggcctcatc ataagatctt agatatgtgt gcagcacctg gctcaaagac 2941 cacacagtta attgaaatgc tacatgccga catgaatgtc ccctttccag agggatttgt 3001 tattgcgaat gatgtggaca acaagcgctg ctacctgctc gtccatcaag ccaagaggct 3061 gagcagcccc tgcatcatgg tggtcaacca tgatgcctcc agcataccca ggctccagat 3121 agatgtggac ggcaggaaag agatcctctt ctatgatcga attttatgtg atgtcccttg 3181 cagtggagac ggcactatga gaaaaaacat tgatgtttgg aaaaagtgga ccaccttaaa 3241 tagcttgcag ctacatggct tacagctgcg gattgcaaca cgcggggctg aacagctggc 3301 tgaaggtgga aggatggtgt attccacgtg ttcactaaac cctattgagg atgaagcagt 3361 catagcatct ttactggaaa aaagtgaagg tgctttggag cttgctgatg tgtctaatga 3421 actgccaggg ctgaagtgga tgcctggaat cacacagtgg aaggtaatga cgaaagatgg 3481 gcagtggttt acagactggg acgctgttcc tcacagcaga cacacccaga tccgacctac 3541 catgttccct ccgaaggacc cagaaaagct gcaggccatg cacctggagc gatgccttag 3601 gatattaccc catcatcaga atactggagg gttttttgtg gcagtattgg tgaaaaaatc 3661 ttcaatgccg tggaataaac gtcagccaaa gcttcagggt aaatctgcag agaccagaga 3721 aagcacacag ctgagccctg cagatctcac agaagggaaa cccacagatc cctctaagct 3781 ggaaagtccg tcattcacag gaactggtga cacagaaata gctcatgcaa ctgaggattt 3841 agagaataat ggcagtaaga aagatggcgt gtgtggtcct cctccatcaa agaaaatgaa 3901 gttatttgga tttaaagaag atccatttgt atttattcct gaagatgacc cattatttcc 3961 acctattgag aaattttatg ctttggatcc ttcattccca aggatgaatt tgttaactcg 4021 gactacagaa gggaagaaaa ggcagctcta catggtttct aaggagttgc ggaatgtgct 4081 gctgaataac agtgagaaga tgaaggttat taacacgggg atcaaagtct ggtgtagaaa 4141 taacagcggt gaagagtttg actgtgcttt ccggctggca caggagggaa tatatacatt 4201 gtatccattt attaactcaa gaattattac tgtatcaatg gaagatgtta agatactgtt 4261 gacccaggaa aatccctttt ttagaaaact cagcagtgag acctacagtc aagcaaagga 4321 cctggcaaag ggaagcatcg tgctgaagta tgaaccagat tctgcgaatc cagacgctct 4381 gcagtgtccc atcgtcttat gcggatggcg gggaaaggcc tccattcgaa cttttgtgcc 4441 caagaatgaa cggcttcatt atctcaggat gatggggctg gaggtattgg gagaaaagaa 4501 gaaggaaggg gttatcctca caaatgagag tgcagccagc accggacagc cagacaatga 4561 cgtgactgag ggacagagag caggagagcc caacagccca gatgcagaag aggccaacag 4621 tccagacgtg acagcaggct gtgacccggc gggggtccat ccaccccggt gagcggccgc 4681 gaaggatctg cgatcgctcc ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc 4741 ccgagaagtt ggggggaggg gtcggcaatt gaacgggtgc ctagagaagg tggcgcgggg 4801 taaactggga aagtgatgtc gtgtactggc tccgcctttt tcccgagggt gggggagaac 4861 cgtatataag tgcagtagtc gccgtgaacg ttctttttcg caacgggttt gccgccagaa 4921 cacagctgaa gcttcgaggg gctcgcatct ctccttcacg cgcccgccgc cctacctgag 4981 gccgccatcc acgccggttg agtcgcgttc tgccgcctcc cgcctgtggt gcctcctgaa 5041 ctgcgtccgc cgtctaggta agtttaaagc tcaggtcgag accgggcctt tgtccggcgc 5101 tcccttggag cctacctaga ctcagccggc tctccacgct ttgcctgacc ctgcttgctc 5161 aactctacgt ctttgtttcg ttttctgttc tgcgccgtta cagatccaag ctgtgaccgg 5221 cgcctacgct agatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc 5281 ccccgggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc 5341 gtcgacccgg accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc 5401 gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg 5461 accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc 5521 gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac 5581 cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc 5641 aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg 5701 cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc 5761 accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag 5821 cccggtgcct gagtcgacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt 5881 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat 5941 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg 6001 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt 6061 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact 6121 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc 6181 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg 6241 tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc 6301 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg 6361 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc 6421 tccccgcctg gtacctttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt 6481 tttaaaagaa aaggggggac tggaagggct aattcactcc caacgaaaat aagatctgct 6541 ttttgcttgt actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta 6601 actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg 6661 tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg 6721 gaaaatctct agcagtagta gttcatgtca tcttattatt cagtatttat aacttgcaaa 6781 gaaatgaata tcagagagtg agaggaactt gtttattgca gcttataatg gttacaaata 6841 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg 6901 tttgtccaaa ctcatcaatg tatcttatca tgtctggctc tagctatccc gcccctaact 6961 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag 7021 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc 7081 ctagactttt gcagagacgg cccaaattcg taatcatggt catagctgtt tcctgtgtga 7141 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc 7201 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc 7261 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc 7321 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt 7381 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca 7441 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa 7501 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat 7561 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc 7621 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc 7681 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt 7741 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac 7801 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg 7861 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca 7921 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc 7981 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa 8041 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa 8101 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac 8161 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta 8221 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt 8281 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata 8341 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc 8401 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac 8461 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag 8521 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac 8581 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc 8641 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg 8701 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc 8761 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct 8821 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc 8881 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc 8941 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc 9001 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc 9061 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca 9121 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt 9181 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt 9241 ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca 9301 ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac 9361 ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat 9421 gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg 9481 cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata 9541 ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc attcgccatt caggctgcgc 9601 aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg 9661 ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt 9721 aaaacgacgg ccagtgccaa gctg //