LOCUS V039454 9871 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V039454 VERSION V039454 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..3897 /label="ANKRD42(NM_001300975)" /note="ANKRD42(NM_001300975)" /gene="ANKRD42" CDS 3904..3969 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 4013..4224 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4237..4505 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4534..4543 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4564..5226 /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 5296..5349 /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 5350..5949 /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 5950..6538 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6612..6845 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6917..7038 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 7078..7213 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(7246..7262) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7270..7286 /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(7294..7324) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7339..7360 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7648..8236) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8407..9267) /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(9268..9372) /gene="bla" /label="AmpR promoter" primer_bind 9846..9862 /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 ccggggtggc caattcaggc 2341 ccctccactt cctctaggga gactgcaaac ccctgttcca ggaagaaggt gcattttggc 2401 agcatacatg atgcagtacg agctggagat gtaaagcagc tttcagaaat agtggtacgt 2461 ggagccagca ttaatgaact tgatgttctc cataagttta cccctttaca ttgggcagca 2521 cattctggaa gtttggagtg tcttcattgg ctgctctggc atggagctga tatcacacac 2581 gtaacaacga gaggttggac agcatctcac atagctgcaa tcaggggtca ggatgcttgt 2641 gtacaggctc ttataatgaa tggagcaaat ctgacagccc aggatgaccg gggatgcact 2701 cctttacatc ttgctgcaac tcatggacat tctttcactt tacaaataat gctccgaagt 2761 ggagtggatc ccagtgtgac tgataagaga gaatggagac ctgtgcatta tgcagctttt 2821 catgggcggc ttggctgctt gcaacttctt gttaaatggg gttgtagcat agaagatgtg 2881 gactacaatg gaaaccttcc agttcactta gcagccatgg aaggccacct tcactgtttc 2941 aaattcctag tcagtagaat gagcagtgcg acgcaagttt taaaagcttt caatgataat 3001 ggagaaaatg tactggattt ggcccagagg ttcttcaagc agaacatttt acagtttatc 3061 cagggggctg agtatgaagg aaaagaccta gaggatcagg aaactttagc atttccaggt 3121 catgtggctg cctttaaggg tgatttgggg atgcttaaga aattagtgga agatggagta 3181 atcaatatta atgagcgtgc tgataatgga tcaactccta tgcataaagc tgctggacaa 3241 ggccacatag agtgtttgca gtggttaatt aaaatgggag cagacagtaa tattaccaac 3301 aaagcagggg agagacccag tgatgtggca aagaggtttg cccatttggc agcagtgaag 3361 ctgttagagg agctacagaa atatgatata gatgacgaaa atgaaattga tgaaaatgat 3421 gtgaaatatt ttataagaca tggtgttgag ggaagcactg atgccaagga tgatttatgt 3481 ctgagtgact tggataaaac agatgccaga atgagagctt acaagaaaat tgtagaattg 3541 agacacctcc tggaaattgc cgagagcaac tataaacact tgggaggcat aacagaagaa 3601 gatttaaagc agaagaaaga acagcttgag tctgaaaaga ccatcaaaga actgcagggc 3661 cagctggagt atgaacgact acgtagagaa aaattagaat gtcagcttga tgaatatcga 3721 gcagaagttg atcaactcag ggaaacactg gaaaaaattc aagtcccaaa ctttgtggct 3781 atggttggtg tcctttttaa tacatttatt tttctcaaga agtatataca agagtggcca 3841 agagtacaag ctttgggctg gggctctttg gacttgaatc ctggctatag tctttttgga 3901 tccgactaca aagaccatga cggtgattat aaagatcatg acatcgacta caaggatgac 3961 gatgacaagt agcggccgcg aaggatctgc gatcgctccg gtgcccgtca gtgggcagag 4021 cgcacatcgc ccacagtccc cgagaagttg gggggagggg tcggcaattg aacgggtgcc 4081 tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg tgtactggct ccgccttttt 4141 cccgagggtg ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc 4201 aacgggtttg ccgccagaac acagctgaag cttcgagggg ctcgcatctc tccttcacgc 4261 gcccgccgcc ctacctgagg ccgccatcca cgccggttga gtcgcgttct gccgcctccc 4321 gcctgtggtg cctcctgaac tgcgtccgcc gtctaggtaa gtttaaagct caggtcgaga 4381 ccgggccttt gtccggcgct cccttggagc ctacctagac tcagccggct ctccacgctt 4441 tgcctgaccc tgcttgctca actctacgtc tttgtttcgt tttctgttct gcgccgttac 4501 agatccaagc tgtgaccggc gcctacgcta gacgccacca tggagagcga cgagagcggc 4561 ctgcccgcca tggagatcga gtgccgcatc accggcaccc tgaacggcgt ggagttcgag 4621 ctggtgggcg gcggagaggg cacccccaag cagggccgca tgaccaacaa gatgaagagc 4681 accaaaggcg ccctgacctt cagcccctac ctgctgagcc acgtgatggg ctacggcttc 4741 taccacttcg gcacctaccc cagcggctac gagaacccct tcctgcacgc catcaacaac 4801 ggcggctaca ccaacacccg catcgagaag tacgaggacg gcggcgtgct gcacgtgagc 4861 ttcagctacc gctacgaggc cggccgcgtg atcggcgact tcaaggtggt gggcaccggc 4921 ttccccgagg acagcgtgat cttcaccgac aagatcatcc gcagcaacgc caccgtggag 4981 cacctgcacc ccatgggcga taacgtgctg gtgggcagct tcgcccgcac cttcagcctg 5041 cgcgacggcg gctactacag cttcgtggtg gacagccaca tgcacttcaa gagcgccatc 5101 caccccagca tcctgcagaa cgggggcccc atgttcgcct tccgccgcgt ggaggagctg 5161 cacagcaaca ccgagctggg catcgtggag taccagcacg ccttcaagac ccccatcgcc 5221 ttcgccagat cccgcgctca gtcgtccaat tctgccgtgg acggcaccgc cggacccggc 5281 tccaccggat ctcgcgaggg cagaggaagt cttctaacat gcggtgacgt ggaggagaat 5341 cccggcccta tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc 5401 agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc 5461 gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc 5521 gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc 5581 acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag 5641 ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg 5701 cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag 5761 ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc 5821 gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc 5881 gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc 5941 ggtgcctgaa atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac 6001 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt 6061 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat 6121 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca 6181 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc 6241 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg 6301 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct 6361 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct 6421 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt 6481 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct 6541 ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga 6601 aaagggggga ctggaagggc taattcactc ccaacgaaga taagatctgc tttttgcttg 6661 tactgggtct ctctggttag accagatctg agcctgggag ctctctggct aactagggaa 6721 cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct 6781 gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc 6841 tagcagtagt agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat 6901 atcagagagt gagaggaact tgtttattgc agcttataat ggttacaaat aaagcaatag 6961 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa 7021 actcatcaat gtatcttatc atgtctggct ctagctatcc cgcccctaac tccgcccatc 7081 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt 7141 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc 7201 ttttttggag gcctagactt ttgcagagac caaattcgta atcatgtcat agctgtttcc 7261 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg 7321 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc 7381 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg 7441 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc 7501 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac 7561 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa 7621 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca 7681 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc 7741 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata 7801 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta 7861 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca 7921 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga 7981 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg 8041 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg 8101 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg 8161 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag 8221 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa 8281 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat 8341 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc 8401 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc 8461 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc 8521 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc 8581 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc 8641 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt 8701 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc 8761 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa 8821 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt 8881 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg 8941 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc 9001 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa 9061 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt 9121 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt 9181 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag 9241 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta 9301 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat 9361 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat 9421 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg 9481 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta 9541 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg 9601 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg 9661 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag 9721 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc 9781 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg 9841 acgttgtaaa acgacggcca gtgccaagct g //