LOCUS V047467 8533 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V047467 VERSION V047467 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..1921 /note="central polypurine tract and central termination sequence of HIV-1" /label="cPPT/CTS" promoter 2011..2214 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 2317..2559 /label="Nedd8(NM_008683)" /note="Nedd8(NM_008683)" /gene="Nedd8" CDS 2566..2631 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 2675..2886 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 2899..3167 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 3196..3205 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 3226..3888 /codon_start=1 /product="green fluorescent protein 2 from Pontellina plumata, also known as ppluGFP2 (Shagin et al., 2004)" /transl_table=1 /translation="PAMEIECRITGTLNGVEFELVGGGEGTPKQGRMTNKMKSTKGALT FSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVLHVSFSYRY EAGRVIGDFKVVGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNVLVGSFARTFSLRDGG YYSFVVDSHMHFKSAIHPSILQNGGPMFAFRRVEELHSNTELGIVEYQHAFKTPIAFA" /label="CopGFP" CDS 3958..4011 /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 Thosea asigna virus capsid protein" /protein_id="" /transl_table=1 /translation="EGRGSLLTCGDVEENPGP" /label="T2A" CDS 4012..4611 /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 4612..5200 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5274..5507 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 5579..5700 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 5740..5875 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(5908..5924) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5932..5948 /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(5956..5986) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6001..6022 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6310..6898) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7069..7929) /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(7930..8034) /gene="bla" /label="AmpR promoter" primer_bind 8508..8524 /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 attacaaaaa ttcaaaattt 1921 tatcgatact agtattatgc ccagtacatg accttatggg actttcctac ttggcagtac 1981 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg 2041 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg 2101 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca 2161 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggttt atataagcag agctcgttta 2221 gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagattc 2281 tagagctagc ctgatacgaa ctcggaattc gccaccatgc taattaaagt gaagacgctg 2341 actgggaagg agattgagat agacatcgaa cccacagaca aggtggagcg aatcaaggag 2401 cgtgtggaag aaaaagaagg gattcccccc cagcagcagc ggctcatcta cagtggcaag 2461 caaatgaatg atgagaagac agcagctgat tacaagattc taggtggttc cgtcctccac 2521 ctggtgttgg ctcttagagg aggaggtggt cttgggcagg gatccgacta caaagaccat 2581 gacggtgatt ataaagatca tgacatcgac tacaaggatg acgatgacaa gtagcggccg 2641 cgaaggatct gcgatcgctc cggtgcccgt cagtgggcag agcgcacatc gcccacagtc 2701 cccgagaagt tggggggagg ggtcggcaat tgaacgggtg cctagagaag gtggcgcggg 2761 gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa 2821 ccgtatataa gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt tgccgccaga 2881 acacagctga agcttcgagg ggctcgcatc tctccttcac gcgcccgccg ccctacctga 2941 ggccgccatc cacgccggtt gagtcgcgtt ctgccgcctc ccgcctgtgg tgcctcctga 3001 actgcgtccg ccgtctaggt aagtttaaag ctcaggtcga gaccgggcct ttgtccggcg 3061 ctcccttgga gcctacctag actcagccgg ctctccacgc tttgcctgac cctgcttgct 3121 caactctacg tctttgtttc gttttctgtt ctgcgccgtt acagatccaa gctgtgaccg 3181 gcgcctacgc tagacgccac catggagagc gacgagagcg gcctgcccgc catggagatc 3241 gagtgccgca tcaccggcac cctgaacggc gtggagttcg agctggtggg cggcggagag 3301 ggcaccccca agcagggccg catgaccaac aagatgaaga gcaccaaagg cgccctgacc 3361 ttcagcccct acctgctgag ccacgtgatg ggctacggct tctaccactt cggcacctac 3421 cccagcggct acgagaaccc cttcctgcac gccatcaaca acggcggcta caccaacacc 3481 cgcatcgaga agtacgagga cggcggcgtg ctgcacgtga gcttcagcta ccgctacgag 3541 gccggccgcg tgatcggcga cttcaaggtg gtgggcaccg gcttccccga ggacagcgtg 3601 atcttcaccg acaagatcat ccgcagcaac gccaccgtgg agcacctgca ccccatgggc 3661 gataacgtgc tggtgggcag cttcgcccgc accttcagcc tgcgcgacgg cggctactac 3721 agcttcgtgg tggacagcca catgcacttc aagagcgcca tccaccccag catcctgcag 3781 aacgggggcc ccatgttcgc cttccgccgc gtggaggagc tgcacagcaa caccgagctg 3841 ggcatcgtgg agtaccagca cgccttcaag acccccatcg ccttcgccag atcccgcgct 3901 cagtcgtcca attctgccgt ggacggcacc gccggacccg gctccaccgg atctcgcgag 3961 ggcagaggaa gtcttctaac atgcggtgac gtggaggaga atcccggccc tatgaccgag 4021 tacaagccca cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt acgcaccctc 4081 gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgatccgga ccgccacatc 4141 gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag 4201 gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa 4261 gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg 4321 gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg 4381 ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc 4441 gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc 4501 gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag 4561 gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg aaatcaacct 4621 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg 4681 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc 4741 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt 4801 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc 4861 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg 4921 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact 4981 gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt 5041 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg 5101 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc 5161 cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggtacctt taagaccaat 5221 gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg 5281 gctaattcac tcccaacgaa gataagatct gctttttgct tgtactgggt ctctctggtt 5341 agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca 5401 ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa 5461 ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta gtagttcatg 5521 tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga gtgagaggaa 5581 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa 5641 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta 5701 tcatgtctgg ctctagctat cccgccccta actccgccca tcccgcccct aactccgccc 5761 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag 5821 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctagac 5881 ttttgcagag accaaattcg taatcatgtc atagctgttt cctgtgtgaa attgttatcc 5941 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta 6001 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa 6061 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat 6121 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg 6181 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc 6241 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt 6301 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag 6361 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc 6421 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc 6481 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt 6541 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt 6601 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc 6661 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa 6721 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa 6781 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg 6841 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga 6901 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg 6961 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg 7021 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt 7081 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact 7141 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat 7201 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg 7261 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg 7321 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat 7381 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc 7441 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt 7501 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc 7561 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga 7621 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc 7681 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa 7741 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta 7801 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg 7861 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg 7921 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat 7981 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt 8041 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa 8101 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct 8161 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag 8221 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc 8281 ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg 8341 cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggga 8401 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc 8461 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc 8521 cagtgccaag ctg //