LOCUS V044498 10186 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V044498 VERSION V044498 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..4212 /label="ZDHHC17(NM_015336)" /note="ZDHHC17(NM_015336)" /gene="ZDHHC17" CDS 4219..4284 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 4328..4539 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4552..4820 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4849..4858 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4879..5541 /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 5611..5664 /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 5665..6264 /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 6265..6853 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6927..7160 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 7232..7353 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 7393..7528 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(7561..7577) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7585..7601 /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(7609..7639) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7654..7675 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7963..8551) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8722..9582) /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(9583..9687) /gene="bla" /label="AmpR promoter" primer_bind 10161..10177 /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 agcgggagga gggatttaac 2341 accaagatgg cggacggccc ggatgagtac gataccgaag cgggctgtgt gccccttctc 2401 cacccagagg aaatcaaacc ccaaagccat tataaccatg gatatggtga acctcttgga 2461 cggaaaactc atattgatga ttacagcaca tgggacatag tcaaggctac acaatatgga 2521 atatatgaac gctgtcgaga attggtggaa gcaggttatg atgtacggca accggacaaa 2581 gaaaatgtta ccctcctcca ttgggctgcc atcaataaca gaatagattt agtcaaatac 2641 tatatttcga aaggtgctat tgtggatcaa cttggagggg acctgaattc aactccattg 2701 cactgggcca caagacaagg ccatctatcc atggttgtgc aactaatgaa atatggtgca 2761 gatccttcat taattgatgg agaaggatgt agctgtattc atctggctgc tcagttcgga 2821 catacctcaa ttgttgctta tctcatagca aaaggacagg atgtagatat gatggatcag 2881 aatggaatga cgcctttaat gtgggcagca tatagaacac atagtgtgga tccaactaga 2941 ttgcttttaa cattcaatgt ttcagttaac cttggtgaca agtatcacaa aaacactgct 3001 ctgcattggg cagtgctagc agggaatacc acagtcatta gccttcttct ggaagctgga 3061 gctaatgttg atgcccagaa tatcaagggc gaatcagcgc ttgatttggc aaaacagaga 3121 aaaaatgtgt ggatgatcaa ccacttacaa gaggcaaggc aagcaaaagg atatgacaat 3181 ccgtccttcc ttagaaagct gaaagctgat aaggaatttc ggcagaaagt aatgttagga 3241 actcctttcc tagttatttg gctggttggg tttatagcag acctaaatat tgattcttgg 3301 ctcattaaag ggctaatgta tggtggtgtt tgggctacag tacagtttct ttcaaaatcc 3361 tttttcgatc attcaatgca tagtgcattg ccccttggga tatatttggc aaccaaattc 3421 tggatgtatg tgacgtggtt cttctggttt tggaatgatc tcaacttttt atttatccat 3481 cttccattcc ttgccaatag tgttgcactt ttctacaatt ttggaaaatc ttggaaatca 3541 gatccaggga ttattaaagc aacagaagag caaaagaaaa agacaatagt tgaacttgca 3601 gagacaggaa gtctggacct cagtatattc tgcagtacct gtttgatacg aaaaccggtg 3661 aggtccaaac attgtggtgt gtgcaaccgc tgtatagcaa aatttgatca tcattgccca 3721 tgggtgggta actgtgtagg tgcaggcaac catagatatt ttatgggcta cctattcttc 3781 ttgcttttta tgatctgctg gatgatttat ggttgtatat cttactgggg actccactgt 3841 gagaccactt acaccaagga tggattttgg acatacatta ctcagattgc cacgtgttca 3901 ccttggatgt tttggatgtt cctgaacagt gttttccact tcatgtgggt ggctgtatta 3961 ctcatgtgtc agatgtacca gatatcatgt ttaggtatta ctacaaatga aagaatgaat 4021 gccaggagat acaagcactt taaagtcaca acaacgtcta ttgaaagccc attcaaccat 4081 ggatgtgtaa gaaatattat agacttcttt gaatttcgat gctgtggcct ctttcgtcct 4141 gttatcgtgg actggaccag gcagtataca atagaatatg accaaatatc aggatctggg 4201 taccagctgg tgggatccga ctacaaagac catgacggtg attataaaga tcatgacatc 4261 gactacaagg atgacgatga caagtagcgg ccgcgaagga tctgcgatcg ctccggtgcc 4321 cgtcagtggg cagagcgcac atcgcccaca gtccccgaga agttgggggg aggggtcggc 4381 aattgaacgg gtgcctagag aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac 4441 tggctccgcc tttttcccga gggtggggga gaaccgtata taagtgcagt agtcgccgtg 4501 aacgttcttt ttcgcaacgg gtttgccgcc agaacacagc tgaagcttcg aggggctcgc 4561 atctctcctt cacgcgcccg ccgccctacc tgaggccgcc atccacgccg gttgagtcgc 4621 gttctgccgc ctcccgcctg tggtgcctcc tgaactgcgt ccgccgtcta ggtaagttta 4681 aagctcaggt cgagaccggg cctttgtccg gcgctccctt ggagcctacc tagactcagc 4741 cggctctcca cgctttgcct gaccctgctt gctcaactct acgtctttgt ttcgttttct 4801 gttctgcgcc gttacagatc caagctgtga ccggcgccta cgctagacgc caccatggag 4861 agcgacgaga gcggcctgcc cgccatggag atcgagtgcc gcatcaccgg caccctgaac 4921 ggcgtggagt tcgagctggt gggcggcgga gagggcaccc ccaagcaggg ccgcatgacc 4981 aacaagatga agagcaccaa aggcgccctg accttcagcc cctacctgct gagccacgtg 5041 atgggctacg gcttctacca cttcggcacc taccccagcg gctacgagaa ccccttcctg 5101 cacgccatca acaacggcgg ctacaccaac acccgcatcg agaagtacga ggacggcggc 5161 gtgctgcacg tgagcttcag ctaccgctac gaggccggcc gcgtgatcgg cgacttcaag 5221 gtggtgggca ccggcttccc cgaggacagc gtgatcttca ccgacaagat catccgcagc 5281 aacgccaccg tggagcacct gcaccccatg ggcgataacg tgctggtggg cagcttcgcc 5341 cgcaccttca gcctgcgcga cggcggctac tacagcttcg tggtggacag ccacatgcac 5401 ttcaagagcg ccatccaccc cagcatcctg cagaacgggg gccccatgtt cgccttccgc 5461 cgcgtggagg agctgcacag caacaccgag ctgggcatcg tggagtacca gcacgccttc 5521 aagaccccca tcgccttcgc cagatcccgc gctcagtcgt ccaattctgc cgtggacggc 5581 accgccggac ccggctccac cggatctcgc gagggcagag gaagtcttct aacatgcggt 5641 gacgtggagg agaatcccgg ccctatgacc gagtacaagc ccacggtgcg cctcgccacc 5701 cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc 5761 acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct gcaagaactc 5821 ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg 5881 gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc 5941 ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggaaggcctc 6001 ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc 6061 gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag 6121 cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag 6181 cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc 6241 atgacccgca agcccggtgc ctgaaatcaa cctctggatt acaaaatttg tgaaagattg 6301 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct 6361 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg 6421 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact 6481 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc 6541 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc 6601 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa 6661 tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc 6721 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg 6781 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg 6841 gccgcctccc cgcctggtac ctttaagacc aatgacttac aaggcagctg tagatcttag 6901 ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac gaagataaga 6961 tctgcttttt gcttgtactg ggtctctctg gttagaccag atctgagcct gggagctctc 7021 tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt 7081 agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc 7141 agtgtggaaa atctctagca gtagtagttc atgtcatctt attattcagt atttataact 7201 tgcaaagaaa tgaatatcag agagtgagag gaacttgttt attgcagctt ataatggtta 7261 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag 7321 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggctctagc tatcccgccc 7381 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc 7441 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag 7501 aagtagtgag gaggcttttt tggaggccta gacttttgca gagaccaaat tcgtaatcat 7561 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc 7621 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc 7681 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat 7741 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac 7801 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt 7861 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca 7921 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc 7981 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact 8041 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct 8101 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag 8161 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca 8221 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa 8281 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc 8341 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag 8401 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg 8461 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca 8521 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc 8581 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag 8641 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata 8701 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat 8761 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg 8821 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc 8881 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc 8941 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc 9001 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc 9061 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc 9121 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa 9181 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat 9241 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata 9301 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca 9361 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag 9421 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc 9481 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc 9541 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata 9601 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta 9661 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta 9721 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg 9781 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt 9841 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg 9901 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt 9961 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg 10021 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct 10081 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg 10141 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctg //