LOCUS V029829 9586 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V029829 VERSION V029829 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..3612 /label="PAICS(NM_001079525)" /note="PAICS(NM_001079525)" /gene="PAICS" CDS 3619..3684 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3728..3939 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3952..4220 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4249..4258 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4279..4941 /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 5011..5064 /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 5065..5664 /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 5665..6253 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6327..6560 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6632..6753 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6793..6928 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6961..6977) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6985..7001 /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(7009..7039) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7054..7075 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7363..7951) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8122..8982) /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(8983..9087) /gene="bla" /label="AmpR promoter" primer_bind 9561..9577 /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 gccaccatgg cgacagctga ggtactgaac 2341 attggtaaaa aattatatga gggtaaaaca aaagaagtct acgaattgtt agacagtcca 2401 ggaaaagtcc tcctgcagtc caaggaccag attacagcag gaaatgcagc tagaaaaaac 2461 cacctggaag gaaaagctgc aatctcaaat aaaatcacca gttgtatttt tcagttatta 2521 caggaagcag ttacctcata taagtcaaat cgtattaaaa ctgccttcac cagaaaatgt 2581 ggggagacag ctttcattgc accgcagtgt gaaatgattc caattgaatg ggtttgcaga 2641 agaatagcaa ctggttcttt tctcaaaaga aatcctggtg tcaaggaagg atataagttt 2701 tacccaccta aagtggagtt gtttttcaag gatgatgcca ataatgaccc acagtggtct 2761 gaggaacagc tgattgctgc aaaattttgc tttgctggac ttcttatagg ccagactgaa 2821 gtggatatca tgagtcatgc tacacaggct atatttgaaa tactggagaa atcctggttg 2881 ccccagaatt gtacactggt tgatatgaag attgaatttg gtgttgatgt aaccaccaaa 2941 gaaattgttc ttgctgatgt tattgacaat gattcctgga gactctggcc atcaggagat 3001 cgaagccaac agaaagacaa acagtcttat cgggacctca aagaagtaac tcctgaaggg 3061 ctccaaatgg taaagaaaaa ctttgagtgg gttgcagaga gagtagagtt gcttttgaaa 3121 tcagaaagtc agtgcagggt tgtagtgttg atgggctcta cttctgatct tggtcactgt 3181 gaaaaaatca agaaggcctg tggaaatttt ggcattccat gtgaacttcg agtaacatct 3241 gcgcataaag gaccagatga aactctgagg attaaagctg agtatgaagg ggatggcatt 3301 cctactgtat ttgtggcagt ggcaggcaga agtaatggtt tgggaccagt gatgtctggg 3361 aacactgcat atccagttat cagctgtcct cccctcacac cagactgggg agttcaggat 3421 gtgtggtctt ctcttcgact acccagtggt cttggctgtt caaccgtact ttctccagaa 3481 ggatcagctc aatttgctgc tcagatattt gggttaagca accatttggt atggagcaaa 3541 ctgcgagcaa gcattttgaa cacatggatt tccttgaagc aggctgacaa gaaaatcaga 3601 gaatgtaatt taggatccga ctacaaagac catgacggtg attataaaga tcatgacatc 3661 gactacaagg atgacgatga caagtagcgg ccgcgaagga tctgcgatcg ctccggtgcc 3721 cgtcagtggg cagagcgcac atcgcccaca gtccccgaga agttgggggg aggggtcggc 3781 aattgaacgg gtgcctagag aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac 3841 tggctccgcc tttttcccga gggtggggga gaaccgtata taagtgcagt agtcgccgtg 3901 aacgttcttt ttcgcaacgg gtttgccgcc agaacacagc tgaagcttcg aggggctcgc 3961 atctctcctt cacgcgcccg ccgccctacc tgaggccgcc atccacgccg gttgagtcgc 4021 gttctgccgc ctcccgcctg tggtgcctcc tgaactgcgt ccgccgtcta ggtaagttta 4081 aagctcaggt cgagaccggg cctttgtccg gcgctccctt ggagcctacc tagactcagc 4141 cggctctcca cgctttgcct gaccctgctt gctcaactct acgtctttgt ttcgttttct 4201 gttctgcgcc gttacagatc caagctgtga ccggcgccta cgctagacgc caccatggag 4261 agcgacgaga gcggcctgcc cgccatggag atcgagtgcc gcatcaccgg caccctgaac 4321 ggcgtggagt tcgagctggt gggcggcgga gagggcaccc ccaagcaggg ccgcatgacc 4381 aacaagatga agagcaccaa aggcgccctg accttcagcc cctacctgct gagccacgtg 4441 atgggctacg gcttctacca cttcggcacc taccccagcg gctacgagaa ccccttcctg 4501 cacgccatca acaacggcgg ctacaccaac acccgcatcg agaagtacga ggacggcggc 4561 gtgctgcacg tgagcttcag ctaccgctac gaggccggcc gcgtgatcgg cgacttcaag 4621 gtggtgggca ccggcttccc cgaggacagc gtgatcttca ccgacaagat catccgcagc 4681 aacgccaccg tggagcacct gcaccccatg ggcgataacg tgctggtggg cagcttcgcc 4741 cgcaccttca gcctgcgcga cggcggctac tacagcttcg tggtggacag ccacatgcac 4801 ttcaagagcg ccatccaccc cagcatcctg cagaacgggg gccccatgtt cgccttccgc 4861 cgcgtggagg agctgcacag caacaccgag ctgggcatcg tggagtacca gcacgccttc 4921 aagaccccca tcgccttcgc cagatcccgc gctcagtcgt ccaattctgc cgtggacggc 4981 accgccggac ccggctccac cggatctcgc gagggcagag gaagtcttct aacatgcggt 5041 gacgtggagg agaatcccgg ccctatgacc gagtacaagc ccacggtgcg cctcgccacc 5101 cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc 5161 acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct gcaagaactc 5221 ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg 5281 gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc 5341 ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggaaggcctc 5401 ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc 5461 gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag 5521 cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag 5581 cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc 5641 atgacccgca agcccggtgc ctgaaatcaa cctctggatt acaaaatttg tgaaagattg 5701 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct 5761 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg 5821 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact 5881 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc 5941 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc 6001 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa 6061 tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc 6121 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg 6181 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg 6241 gccgcctccc cgcctggtac ctttaagacc aatgacttac aaggcagctg tagatcttag 6301 ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac gaagataaga 6361 tctgcttttt gcttgtactg ggtctctctg gttagaccag atctgagcct gggagctctc 6421 tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt 6481 agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc 6541 agtgtggaaa atctctagca gtagtagttc atgtcatctt attattcagt atttataact 6601 tgcaaagaaa tgaatatcag agagtgagag gaacttgttt attgcagctt ataatggtta 6661 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag 6721 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggctctagc tatcccgccc 6781 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc 6841 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag 6901 aagtagtgag gaggcttttt tggaggccta gacttttgca gagaccaaat tcgtaatcat 6961 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc 7021 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc 7081 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat 7141 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac 7201 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt 7261 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca 7321 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc 7381 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact 7441 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct 7501 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag 7561 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca 7621 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa 7681 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc 7741 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag 7801 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg 7861 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca 7921 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc 7981 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag 8041 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata 8101 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat 8161 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg 8221 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc 8281 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc 8341 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc 8401 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc 8461 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc 8521 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa 8581 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat 8641 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata 8701 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca 8761 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag 8821 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc 8881 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc 8941 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata 9001 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta 9061 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta 9121 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg 9181 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt 9241 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg 9301 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt 9361 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg 9421 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct 9481 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg 9541 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctg //