LOCUS V042258 9913 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V042258 VERSION V042258 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..3939 /label="NR5A2(NM_205860)" /note="NR5A2(NM_205860)" /gene="NR5A2" CDS 3946..4011 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 4055..4266 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4279..4547 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4576..4585 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4606..5268 /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 5338..5391 /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 5392..5991 /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 5992..6580 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6654..6887 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6959..7080 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 7120..7255 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(7288..7304) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7312..7328 /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(7336..7366) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7381..7402 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7690..8278) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8449..9309) /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(9310..9414) /gene="bla" /label="AmpR promoter" primer_bind 9888..9904 /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 gccaccatgt cttctaattc agatactggg 2341 gatttacaag agtctttaaa gcacggactt acacctattg gtgctgggct tccggaccga 2401 cacggatccc ccatccccgc ccgcggtcgc cttgtcatgc tgcccaaagt ggagacggaa 2461 gccctgggac tggctcgatc gcatggggaa cagggccaga tgccggaaaa catgcaagtg 2521 tctcaattta aaatggtgaa ttactcctat gatgaagatc tggaagagct ttgtcccgtg 2581 tgtggagata aagtgtctgg gtaccattat gggctcctca cctgtgaaag ctgcaaggga 2641 ttttttaagc gaacagtcca aaataataaa aggtacacat gtatagaaaa ccagaactgc 2701 caaattgaca aaacacagag aaagcgttgt ccttactgtc gttttcaaaa atgtctaagt 2761 gttggaatga agctagaagc tgtaagggcc gaccgaatgc gtggaggaag gaataagttt 2821 gggccaatgt acaagagaga cagggccctg aagcaacaga aaaaagccct catccgagcc 2881 aatggactta agctagaagc catgtctcag gtgatccaag ctatgccctc tgacctgacc 2941 atttcctctg caattcaaaa catccactct gcctccaaag gcctacctct gaaccatgct 3001 gccttgcctc ctacagacta tgacagaagt ccctttgtaa catcccccat tagcatgaca 3061 atgccccctc acggcagcct gcaaggttac caaacatatg gccactttcc tagccgggcc 3121 atcaagtctg agtacccaga cccctatacc agctcacccg agtccataat gggctattca 3181 tatatggata gttaccagac gagctctcca gcaagcatcc cacatctgat actggaactt 3241 ttgaagtgtg agccagatga gcctcaagtc caggctaaaa tcatggccta tttgcagcaa 3301 gagcaggcta accgaagcaa gcacgaaaag ctgagcacct ttgggcttat gtgcaaaatg 3361 gcagatcaaa ctctcttctc cattgtcgag tgggccagga gtagtatctt cttcagagaa 3421 cttaaggttg atgaccaaat gaagctgctt cagaactgct ggagtgagct cttaatcctc 3481 gaccacattt accgacaagt ggtacatgga aaggaaggat ccatcttcct ggttactggg 3541 caacaagtgg actattccat aatagcatca caagccggag ccaccctcaa caacctcatg 3601 agtcatgcac aggagttagt ggcaaaactt cgttctctcc agtttgatca acgagagttc 3661 gtatgtctga aattcttggt gctctttagt ttagatgtca aaaaccttga aaacttccag 3721 ctggtagaag gtgtccagga acaagtcaat gccgccctgc tggactacac aatgtgtaac 3781 tacccgcagc agacagagaa atttggacag ctacttcttc gactacccga aatccgggcc 3841 atcagtatgc aggctgaaga atacctctac tacaagcacc tgaacgggga tgtgccctat 3901 aataaccttc tcattgaaat gttgcatgcc aaaagagcag gatccgacta caaagaccat 3961 gacggtgatt ataaagatca tgacatcgac tacaaggatg acgatgacaa gtagcggccg 4021 cgaaggatct gcgatcgctc cggtgcccgt cagtgggcag agcgcacatc gcccacagtc 4081 cccgagaagt tggggggagg ggtcggcaat tgaacgggtg cctagagaag gtggcgcggg 4141 gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa 4201 ccgtatataa gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt tgccgccaga 4261 acacagctga agcttcgagg ggctcgcatc tctccttcac gcgcccgccg ccctacctga 4321 ggccgccatc cacgccggtt gagtcgcgtt ctgccgcctc ccgcctgtgg tgcctcctga 4381 actgcgtccg ccgtctaggt aagtttaaag ctcaggtcga gaccgggcct ttgtccggcg 4441 ctcccttgga gcctacctag actcagccgg ctctccacgc tttgcctgac cctgcttgct 4501 caactctacg tctttgtttc gttttctgtt ctgcgccgtt acagatccaa gctgtgaccg 4561 gcgcctacgc tagacgccac catggagagc gacgagagcg gcctgcccgc catggagatc 4621 gagtgccgca tcaccggcac cctgaacggc gtggagttcg agctggtggg cggcggagag 4681 ggcaccccca agcagggccg catgaccaac aagatgaaga gcaccaaagg cgccctgacc 4741 ttcagcccct acctgctgag ccacgtgatg ggctacggct tctaccactt cggcacctac 4801 cccagcggct acgagaaccc cttcctgcac gccatcaaca acggcggcta caccaacacc 4861 cgcatcgaga agtacgagga cggcggcgtg ctgcacgtga gcttcagcta ccgctacgag 4921 gccggccgcg tgatcggcga cttcaaggtg gtgggcaccg gcttccccga ggacagcgtg 4981 atcttcaccg acaagatcat ccgcagcaac gccaccgtgg agcacctgca ccccatgggc 5041 gataacgtgc tggtgggcag cttcgcccgc accttcagcc tgcgcgacgg cggctactac 5101 agcttcgtgg tggacagcca catgcacttc aagagcgcca tccaccccag catcctgcag 5161 aacgggggcc ccatgttcgc cttccgccgc gtggaggagc tgcacagcaa caccgagctg 5221 ggcatcgtgg agtaccagca cgccttcaag acccccatcg ccttcgccag atcccgcgct 5281 cagtcgtcca attctgccgt ggacggcacc gccggacccg gctccaccgg atctcgcgag 5341 ggcagaggaa gtcttctaac atgcggtgac gtggaggaga atcccggccc tatgaccgag 5401 tacaagccca cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt acgcaccctc 5461 gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgatccgga ccgccacatc 5521 gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag 5581 gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa 5641 gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg 5701 gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg 5761 ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc 5821 gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc 5881 gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag 5941 gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg aaatcaacct 6001 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg 6061 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc 6121 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt 6181 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc 6241 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg 6301 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact 6361 gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt 6421 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg 6481 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc 6541 cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggtacctt taagaccaat 6601 gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg 6661 gctaattcac tcccaacgaa gataagatct gctttttgct tgtactgggt ctctctggtt 6721 agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca 6781 ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa 6841 ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta gtagttcatg 6901 tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga gtgagaggaa 6961 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa 7021 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta 7081 tcatgtctgg ctctagctat cccgccccta actccgccca tcccgcccct aactccgccc 7141 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag 7201 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctagac 7261 ttttgcagag accaaattcg taatcatgtc atagctgttt cctgtgtgaa attgttatcc 7321 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta 7381 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa 7441 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat 7501 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg 7561 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc 7621 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt 7681 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag 7741 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc 7801 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc 7861 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt 7921 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt 7981 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc 8041 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa 8101 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa 8161 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg 8221 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga 8281 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg 8341 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg 8401 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt 8461 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact 8521 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat 8581 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg 8641 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg 8701 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat 8761 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc 8821 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt 8881 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc 8941 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga 9001 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc 9061 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa 9121 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta 9181 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg 9241 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg 9301 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat 9361 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt 9421 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa 9481 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct 9541 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag 9601 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc 9661 ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg 9721 cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggga 9781 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc 9841 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc 9901 cagtgccaag ctg //