LOCUS V022326 9550 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V022326 VERSION V022326 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 2303..3664 /label="DAPK3(NM_001348)" /note="DAPK3(NM_001348)" /gene="DAPK3" promoter 3709..3920 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3933..4201 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4230..4239 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4260..4922 /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 4992..5045 /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 5046..5645 /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 5646..6233 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6307..6540 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6612..6733 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6752..6887 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6925..6941) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6949..6965 /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(6973..7003) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7018..7039 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7327..7915) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8086..8946) /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(8947..9051) /gene="bla" /label="AmpR promoter" primer_bind 9525..9541 /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 gaattcgcca ccatgtccac gttcaggcag gaggacgtgg aggaccatta 2341 tgagatgggg gaggagctgg gcagcggcca gtttgcgatc gtgcggaagt gccggcagaa 2401 gggcacgggc aaggagtacg cagccaagtt catcaagaag cgccgcctgt catccagccg 2461 gcgtggggtg agccgggagg agatcgagcg ggaggtgaac atcctgcggg agatccggca 2521 ccccaacatc atcaccctgc acgacatctt cgagaacaag acggacgtgg tcctcatcct 2581 ggagctggtc tctggcgggg agctctttga cttcctggcg gagaaggagt cgctgacgga 2641 ggacgaggcc acccagttcc tcaagcagat cctggacggc gttcactacc tgcactctaa 2701 gcgcatcgca cactttgacc tgaagccgga aaacatcatg ctgctggaca agaacgtgcc 2761 caacccacga atcaagctca tcgacttcgg catcgcgcac aagatcgagg cggggaacga 2821 gttcaagaac atcttcggca ccccggagtt tgtggcccca gagattgtga actatgagcc 2881 gctgggcctg gaggcggaca tgtggagcat cggtgtcatc acctatatcc tcctgagcgg 2941 tgcatccccg ttcctgggcg agaccaagca ggagacgctc accaacatct cagccgtgaa 3001 ctacgacttc gacgaggagt acttcagcaa caccagcgag ctggccaagg acttcattcg 3061 ccggctgctc gtcaaagatc ccaagcggag aatgaccatt gcccagagcc tggaacattc 3121 ctggattaag gcgatccggc ggcggaacgt gcgtggtgag gacagcggcc gcaagcccga 3181 gcggcggcgc ctgaagacca cgcgtctgaa ggagtacacc atcaagtcgc actccagctt 3241 gccgcccaac aacagctacg ccgacttcga gcgcttctcc aaggtgctgg aggaggcggc 3301 ggccgccgag gagggcctgc gcgagctgca gcgcagccgg cggctctgcc acgaggacgt 3361 ggaggcgctg gccgccatct acgaggagaa ggaggcctgg taccgcgagg agagcgacag 3421 cctgggccag gacctgcgga ggctacggca ggagctgctc aagaccgagg cgctcaagcg 3481 gcaggcgcag gaggaggcca agggcgcgct gctggggacc agcggcctca agcgccgctt 3541 cagccgcctg gagaaccgct acgaggcgct ggccaagcaa gtagcctccg agatgcgctt 3601 cgtgcaggac ctcgtgcgcg ccctggagca ggagaagctg cagggcgtgg agtgcgggct 3661 gcgctaggcg gccgcgaagg atctgcgatc gctccggtgc ccgtcagtgg gcagagcgca 3721 catcgcccac agtccccgag aagttggggg gaggggtcgg caattgaacg ggtgcctaga 3781 gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg 3841 agggtggggg agaaccgtat ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg 3901 ggtttgccgc cagaacacag ctgaagcttc gaggggctcg catctctcct tcacgcgccc 3961 gccgccctac ctgaggccgc catccacgcc ggttgagtcg cgttctgccg cctcccgcct 4021 gtggtgcctc ctgaactgcg tccgccgtct aggtaagttt aaagctcagg tcgagaccgg 4081 gcctttgtcc ggcgctccct tggagcctac ctagactcag ccggctctcc acgctttgcc 4141 tgaccctgct tgctcaactc tacgtctttg tttcgttttc tgttctgcgc cgttacagat 4201 ccaagctgtg accggcgcct acgctagacg ccaccatgga gagcgacgag agcggcctgc 4261 ccgccatgga gatcgagtgc cgcatcaccg gcaccctgaa cggcgtggag ttcgagctgg 4321 tgggcggcgg agagggcacc cccaagcagg gccgcatgac caacaagatg aagagcacca 4381 aaggcgccct gaccttcagc ccctacctgc tgagccacgt gatgggctac ggcttctacc 4441 acttcggcac ctaccccagc ggctacgaga accccttcct gcacgccatc aacaacggcg 4501 gctacaccaa cacccgcatc gagaagtacg aggacggcgg cgtgctgcac gtgagcttca 4561 gctaccgcta cgaggccggc cgcgtgatcg gcgacttcaa ggtggtgggc accggcttcc 4621 ccgaggacag cgtgatcttc accgacaaga tcatccgcag caacgccacc gtggagcacc 4681 tgcaccccat gggcgataac gtgctggtgg gcagcttcgc ccgcaccttc agcctgcgcg 4741 acggcggcta ctacagcttc gtggtggaca gccacatgca cttcaagagc gccatccacc 4801 ccagcatcct gcagaacggg ggccccatgt tcgccttccg ccgcgtggag gagctgcaca 4861 gcaacaccga gctgggcatc gtggagtacc agcacgcctt caagaccccc atcgccttcg 4921 ccagatcccg cgctcagtcg tccaattctg ccgtggacgg caccgccgga cccggctcca 4981 ccggatctcg cgagggcaga ggaagtcttc taacatgcgg tgacgtggag gagaatcccg 5041 gccctatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtccccaggg 5101 ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgatc 5161 cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc 5221 tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc 5281 cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga 5341 gcggttcccg gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca 5401 aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc 5461 tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct 5521 tcctggagac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca 5581 ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg 5641 cctgaaatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg 5701 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt 5761 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg 5821 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc 5881 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc 5941 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc 6001 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc 6061 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg 6121 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc 6181 gtctccgcct tcgccctcag acgagtcgga tctccctttg gccgcctccc cgcctggtac 6241 ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg 6301 ggggactgga agggctaatt cactcccaac gaaaataaga tctgcttttt gcttgtactg 6361 ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac 6421 tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt 6481 gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca 6541 gtagtagttc atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag 6601 agagtgagag gaacttgttt attgcagctt ataatggtta caaataaagc aatagcatca 6661 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca 6721 tcaatgtatc ttatcatgtc tggctctagc tatcccgccc ctaactccgc ccagttccgc 6781 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc 6841 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag acttttgcag 6901 agacggccca aattcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct 6961 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg 7021 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct 7081 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg 7141 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc 7201 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg 7261 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct 7321 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca 7381 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct 7441 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc 7501 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt 7561 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc 7621 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc 7681 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg 7741 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc 7801 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag 7861 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga 7921 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat 7981 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag 8041 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat 8101 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc 8161 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat 8221 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag 8281 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg 8341 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc 8401 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca 8461 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg 8521 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc 8581 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta 8641 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc 8701 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg 8761 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc 8821 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc 8881 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat 8941 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag 9001 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc 9061 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa 9121 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg 9181 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca 9241 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc 9301 atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt 9361 aaggagaaaa taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg 9421 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag 9481 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag 9541 tgccaagctg //