LOCUS V029569 9856 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V029569 VERSION V029569 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..3882 /label="Trim65(NM_178802)" /note="Trim65(NM_178802)" /gene="Trim65" CDS 3889..3954 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3998..4209 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4222..4490 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4519..4528 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4549..5211 /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 5281..5334 /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 5335..5934 /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 5935..6523 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6597..6830 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6902..7023 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 7063..7198 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(7231..7247) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7255..7271 /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(7279..7309) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7324..7345 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7633..8221) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8392..9252) /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(9253..9357) /gene="bla" /label="AmpR promoter" primer_bind 9831..9847 /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 ccgctcagtt gctggaggag 2341 gacgtggtga cttgctccat ctgcttaggt cgctaccggg acccggtgac gctgccctgc 2401 gggcacagct tctgcgggaa ctgcatccag gattcatggc gttcctgcga aaagtcgtgt 2461 ccggaatgtc gccagccctt ccccgagggc gccaagctgt cccgcaacgt gaagatgagc 2521 accttgctgc aggcgctgcc cgttctgccg gcgccgcccg ccgtgactcc gcgccgggac 2581 tccgccacaa gccacagcgc tcgctgcctg cgccacggga ggccgctcga gttcttctgt 2641 cgcacggagg gcctctgcgt gtgcagcgcg tgcacggtgc acgactgcag ccaccacgag 2701 cgggcgctgc tggacgtgga gcgccgcgtg cgcgaggacc agctgagagc gagggtactg 2761 gttacccagc agcaggttgc ccaggcagag acccagcttc aggagctgca ggagcaacgc 2821 agtcggattg agagttcagc ctgcaccctg gcctcggtgg tctccaggag attcagtagc 2881 ctgcttcagg cgctggagaa gcagcaggcc tcaacactga gtgatataga ggtagccaag 2941 aagcaggcgc tgggccaagt tctgaatgag aaacagcggc tgactgacca cctaagggcc 3001 ctgtctcagt atgatcagag tgtccaggat ctcctggcac aggcggatga ctgcatcttc 3061 ttccaggaat tacagcagct ccctgagccc acagagtccc tcgggccact gacctctcca 3121 cagtggaatg aagaacagca gctgagcaac gtgaaccagc tgcttagccc cctgtgtgag 3181 ctcctccttg aagagaagag cctccccaag gtggcagcca aggctgcaga tgctggccct 3241 ttggagaccc tgggtcccct ggcaccagtc cccagcgcag tgtgtccact gaggaagaag 3301 ctctggcaga attatcgcaa tctgaccttt gacccagaga ccgccaacca gtatttgcac 3361 ttgtctcaca aggaccagcg ggtgacacac catttccagg cccagggccc agccaagtcg 3421 ggcagcttcg agctgtggca ggtgcagtgt acccagagtt ttcagactgg acaacattac 3481 tgggaggtac gtgtctctga tcactctgtc actctgggcg tcacctaccc aaagctatca 3541 cgccaaaagc tggggaccca cacggataac atcggccgtg gaccctgctc ctggggcctc 3601 tgcatccagg aggacagtat gcaggcctgg cacaatggca aatctcagcg cctgcgtgga 3661 ttgcccggac aactcctggg cgtagatttg aacttgacct ctggctgcct caccttctac 3721 agcctggagc cacgcacaca gccgctgcac actttctatg cagtgttcag ccagcctctc 3781 ttccctgtct tctggctcct tgaggggagg acccttactc tgtgccatca gcctgaggcc 3841 acgctccctg caaggcccca ggaagaggcc acagcaccca gcggatccga ctacaaagac 3901 catgacggtg attataaaga tcatgacatc gactacaagg atgacgatga caagtagcgg 3961 ccgcgaagga tctgcgatcg ctccggtgcc cgtcagtggg cagagcgcac atcgcccaca 4021 gtccccgaga agttgggggg aggggtcggc aattgaacgg gtgcctagag aaggtggcgc 4081 ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc tttttcccga gggtggggga 4141 gaaccgtata taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg gtttgccgcc 4201 agaacacagc tgaagcttcg aggggctcgc atctctcctt cacgcgcccg ccgccctacc 4261 tgaggccgcc atccacgccg gttgagtcgc gttctgccgc ctcccgcctg tggtgcctcc 4321 tgaactgcgt ccgccgtcta ggtaagttta aagctcaggt cgagaccggg cctttgtccg 4381 gcgctccctt ggagcctacc tagactcagc cggctctcca cgctttgcct gaccctgctt 4441 gctcaactct acgtctttgt ttcgttttct gttctgcgcc gttacagatc caagctgtga 4501 ccggcgccta cgctagacgc caccatggag agcgacgaga gcggcctgcc cgccatggag 4561 atcgagtgcc gcatcaccgg caccctgaac ggcgtggagt tcgagctggt gggcggcgga 4621 gagggcaccc ccaagcaggg ccgcatgacc aacaagatga agagcaccaa aggcgccctg 4681 accttcagcc cctacctgct gagccacgtg atgggctacg gcttctacca cttcggcacc 4741 taccccagcg gctacgagaa ccccttcctg cacgccatca acaacggcgg ctacaccaac 4801 acccgcatcg agaagtacga ggacggcggc gtgctgcacg tgagcttcag ctaccgctac 4861 gaggccggcc gcgtgatcgg cgacttcaag gtggtgggca ccggcttccc cgaggacagc 4921 gtgatcttca ccgacaagat catccgcagc aacgccaccg tggagcacct gcaccccatg 4981 ggcgataacg tgctggtggg cagcttcgcc cgcaccttca gcctgcgcga cggcggctac 5041 tacagcttcg tggtggacag ccacatgcac ttcaagagcg ccatccaccc cagcatcctg 5101 cagaacgggg gccccatgtt cgccttccgc cgcgtggagg agctgcacag caacaccgag 5161 ctgggcatcg tggagtacca gcacgccttc aagaccccca tcgccttcgc cagatcccgc 5221 gctcagtcgt ccaattctgc cgtggacggc accgccggac ccggctccac cggatctcgc 5281 gagggcagag gaagtcttct aacatgcggt gacgtggagg agaatcccgg ccctatgacc 5341 gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tccccagggc cgtacgcacc 5401 ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac 5461 atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc 5521 aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc 5581 gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg 5641 ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg 5701 tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc 5761 gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc 5821 tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc 5881 gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctgaaatcaa 5941 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt 6001 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct 6061 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc 6121 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg 6181 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc 6241 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc 6301 actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt 6361 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca 6421 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt 6481 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggtac ctttaagacc 6541 aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg ggggactgga 6601 agggctaatt cactcccaac gaagataaga tctgcttttt gcttgtactg ggtctctctg 6661 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc 6721 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg 6781 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gtagtagttc 6841 atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag agagtgagag 6901 gaacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac 6961 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc 7021 ttatcatgtc tggctctagc tatcccgccc ctaactccgc ccatcccgcc cctaactccg 7081 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc 7141 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta 7201 gacttttgca gagaccaaat tcgtaatcat gtcatagctg tttcctgtgt gaaattgtta 7261 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc 7321 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg 7381 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg 7441 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg 7501 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa 7561 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc 7621 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc 7681 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag 7741 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct 7801 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta 7861 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc 7921 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc 7981 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt 8041 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct 8101 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc 8161 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca 8221 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta 8281 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa 8341 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg 8401 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg 8461 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc 8521 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc 8581 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa 8641 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc 8701 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg 8761 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc 8821 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat 8881 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg 8941 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc 9001 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg 9061 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat 9121 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg 9181 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg 9241 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct 9301 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac 9361 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta 9421 taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa 9481 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag 9541 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta 9601 tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag 9661 atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg 9721 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc 9781 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac 9841 ggccagtgcc aagctg //