LOCUS V047587 8956 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V047587 VERSION V047587 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..2982 /label="GSTA1(NM_145740)" /note="GSTA1(NM_145740)" /gene="GSTA1" CDS 2989..3054 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3098..3309 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3322..3590 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 3619..3628 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 3649..4311 /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 4381..4434 /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 4435..5034 /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 5035..5623 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5697..5930 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6002..6123 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6163..6298 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6331..6347) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6355..6371 /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(6379..6409) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6424..6445 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6733..7321) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7492..8352) /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(8353..8457) /gene="bla" /label="AmpR promoter" primer_bind 8931..8947 /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 cagagaagcc caagctccac 2341 tacttcaatg cacggggcag aatggagtcc acccggtggc tcctggctgc agctggagta 2401 gagtttgaag agaaatttat aaaatctgca gaagatttgg acaagttaag aaatgatgga 2461 tatttgatgt tccagcaagt gccaatggtt gagattgatg ggatgaagct ggtgcagacc 2521 agagccattc tcaactacat tgccagcaaa tacaacctct atgggaaaga cataaaggag 2581 agagccctga ttgatatgta tatagaaggt atagcagatt tgggtgaaat gatcctcctt 2641 ctgcccgtat gtccacctga ggaaaaagat gccaagcttg ccttgatcaa agagaaaata 2701 aaaaatcgct acttccctgc ctttgaaaaa gtcttaaaga gccatggaca agactacctt 2761 gttggcaaca agctgagccg ggctgacatt catctggtgg aacttctcta ctacgtcgag 2821 gagcttgact ccagtcttat ctccagcttc cctctgctga aggccctgaa aaccagaatc 2881 agcaacctgc ccacagtgaa gaagtttcta cagcctggca gcccaaggaa gcctcccatg 2941 gatgagaaat ctttagaaga agcaaggaag attttcaggt ttggatccga ctacaaagac 3001 catgacggtg attataaaga tcatgacatc gactacaagg atgacgatga caagtagcgg 3061 ccgcgaagga tctgcgatcg ctccggtgcc cgtcagtggg cagagcgcac atcgcccaca 3121 gtccccgaga agttgggggg aggggtcggc aattgaacgg gtgcctagag aaggtggcgc 3181 ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc tttttcccga gggtggggga 3241 gaaccgtata taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg gtttgccgcc 3301 agaacacagc tgaagcttcg aggggctcgc atctctcctt cacgcgcccg ccgccctacc 3361 tgaggccgcc atccacgccg gttgagtcgc gttctgccgc ctcccgcctg tggtgcctcc 3421 tgaactgcgt ccgccgtcta ggtaagttta aagctcaggt cgagaccggg cctttgtccg 3481 gcgctccctt ggagcctacc tagactcagc cggctctcca cgctttgcct gaccctgctt 3541 gctcaactct acgtctttgt ttcgttttct gttctgcgcc gttacagatc caagctgtga 3601 ccggcgccta cgctagacgc caccatggag agcgacgaga gcggcctgcc cgccatggag 3661 atcgagtgcc gcatcaccgg caccctgaac ggcgtggagt tcgagctggt gggcggcgga 3721 gagggcaccc ccaagcaggg ccgcatgacc aacaagatga agagcaccaa aggcgccctg 3781 accttcagcc cctacctgct gagccacgtg atgggctacg gcttctacca cttcggcacc 3841 taccccagcg gctacgagaa ccccttcctg cacgccatca acaacggcgg ctacaccaac 3901 acccgcatcg agaagtacga ggacggcggc gtgctgcacg tgagcttcag ctaccgctac 3961 gaggccggcc gcgtgatcgg cgacttcaag gtggtgggca ccggcttccc cgaggacagc 4021 gtgatcttca ccgacaagat catccgcagc aacgccaccg tggagcacct gcaccccatg 4081 ggcgataacg tgctggtggg cagcttcgcc cgcaccttca gcctgcgcga cggcggctac 4141 tacagcttcg tggtggacag ccacatgcac ttcaagagcg ccatccaccc cagcatcctg 4201 cagaacgggg gccccatgtt cgccttccgc cgcgtggagg agctgcacag caacaccgag 4261 ctgggcatcg tggagtacca gcacgccttc aagaccccca tcgccttcgc cagatcccgc 4321 gctcagtcgt ccaattctgc cgtggacggc accgccggac ccggctccac cggatctcgc 4381 gagggcagag gaagtcttct aacatgcggt gacgtggagg agaatcccgg ccctatgacc 4441 gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tccccagggc cgtacgcacc 4501 ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac 4561 atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc 4621 aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc 4681 gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg 4741 ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg 4801 tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc 4861 gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc 4921 tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc 4981 gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctgaaatcaa 5041 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt 5101 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct 5161 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc 5221 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg 5281 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc 5341 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc 5401 actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt 5461 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca 5521 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt 5581 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggtac ctttaagacc 5641 aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg ggggactgga 5701 agggctaatt cactcccaac gaagataaga tctgcttttt gcttgtactg ggtctctctg 5761 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc 5821 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg 5881 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gtagtagttc 5941 atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag agagtgagag 6001 gaacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac 6061 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc 6121 ttatcatgtc tggctctagc tatcccgccc ctaactccgc ccatcccgcc cctaactccg 6181 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc 6241 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta 6301 gacttttgca gagaccaaat tcgtaatcat gtcatagctg tttcctgtgt gaaattgtta 6361 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc 6421 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg 6481 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg 6541 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg 6601 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa 6661 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc 6721 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc 6781 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag 6841 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct 6901 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta 6961 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc 7021 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc 7081 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt 7141 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct 7201 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc 7261 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca 7321 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta 7381 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa 7441 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg 7501 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg 7561 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc 7621 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc 7681 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa 7741 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc 7801 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg 7861 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc 7921 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat 7981 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg 8041 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc 8101 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg 8161 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat 8221 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg 8281 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg 8341 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct 8401 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac 8461 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta 8521 taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa 8581 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag 8641 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta 8701 tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag 8761 atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg 8821 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc 8881 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac 8941 ggccagtgcc aagctg //