LOCUS V043335 9346 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V043335 VERSION V043335 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..3372 /label="OTULIN(NM_138348)" /note="OTULIN(NM_138348)" /gene="OTULIN" CDS 3379..3444 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3488..3699 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3712..3980 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4009..4018 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4039..4701 /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 4771..4824 /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 4825..5424 /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 5425..6013 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6087..6320 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6392..6513 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6553..6688 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6721..6737) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6745..6761 /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(6769..6799) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6814..6835 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7123..7711) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7882..8742) /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(8743..8847) /gene="bla" /label="AmpR promoter" primer_bind 9321..9337 /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 gccaccatga gtcgggggac tatgccccag 2341 cccgaagcgt ggccaggcgc gagctgcgcc gagacgccgg cgcgggaggc ggcggccacg 2401 gcgcgggacg gcgggaaggc ggcggccagc gggcagccgc ggcccgagat gcagtgcccg 2461 gccgagcatg aggaggacat gtaccgtgct gcagatgaaa tagaaaagga gaaagaattg 2521 cttatacatg aaagaggggc atcagaaccg agattaagcg tagctcctga aatggatatc 2581 atggactact gcaaaaaaga atggagagga aatacacaga aagcaacgtg tatgaaaatg 2641 ggctatgaag aggtttctca gaagttcacc tccatacggc gagtccgtgg tgataattac 2701 tgtgcactga gggccacgct gttccaggcc atgagccagg ctgtggggct gccgccctgg 2761 ctgcaggacc cggagctcat gctgttacca gaaaaactca taagcaaata caactggatc 2821 aagcaatgga aacttggact gaaatttgat gggaagaatg aggacctggt tgataaaatt 2881 aaagagtccc ttactctgct gaggaagaag tgggcaggct tggctgaaat gagaactgct 2941 gaagcaagac agatagcttg tgatgaacta ttcacaaatg aggcggagga atatagcctc 3001 tatgaagctg taaaatttct aatgctaaac agagccattg aactatataa tgataaagag 3061 aaaggaaagg aagtaccatt tttctctgtg cttctgtttg ctcgggacac atcaaatgac 3121 ccaggacagc ttctgaggaa ccacctcaac caggtgggac acactggtgg tcttgaacag 3181 gttgaaatgt tccttcttgc ctatgctgtg cgccacacca tccaggtgta ccggctctcc 3241 aagtacaaca cggaagaatt catcacagtc taccccaccg acccacccaa ggactggcca 3301 gtggtaacgc tcattgctga ggacgatcgg cactataaca tccccgtcag agtgtgtgag 3361 gagaccagtc taggatccga ctacaaagac catgacggtg attataaaga tcatgacatc 3421 gactacaagg atgacgatga caagtagcgg ccgcgaagga tctgcgatcg ctccggtgcc 3481 cgtcagtggg cagagcgcac atcgcccaca gtccccgaga agttgggggg aggggtcggc 3541 aattgaacgg gtgcctagag aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac 3601 tggctccgcc tttttcccga gggtggggga gaaccgtata taagtgcagt agtcgccgtg 3661 aacgttcttt ttcgcaacgg gtttgccgcc agaacacagc tgaagcttcg aggggctcgc 3721 atctctcctt cacgcgcccg ccgccctacc tgaggccgcc atccacgccg gttgagtcgc 3781 gttctgccgc ctcccgcctg tggtgcctcc tgaactgcgt ccgccgtcta ggtaagttta 3841 aagctcaggt cgagaccggg cctttgtccg gcgctccctt ggagcctacc tagactcagc 3901 cggctctcca cgctttgcct gaccctgctt gctcaactct acgtctttgt ttcgttttct 3961 gttctgcgcc gttacagatc caagctgtga ccggcgccta cgctagacgc caccatggag 4021 agcgacgaga gcggcctgcc cgccatggag atcgagtgcc gcatcaccgg caccctgaac 4081 ggcgtggagt tcgagctggt gggcggcgga gagggcaccc ccaagcaggg ccgcatgacc 4141 aacaagatga agagcaccaa aggcgccctg accttcagcc cctacctgct gagccacgtg 4201 atgggctacg gcttctacca cttcggcacc taccccagcg gctacgagaa ccccttcctg 4261 cacgccatca acaacggcgg ctacaccaac acccgcatcg agaagtacga ggacggcggc 4321 gtgctgcacg tgagcttcag ctaccgctac gaggccggcc gcgtgatcgg cgacttcaag 4381 gtggtgggca ccggcttccc cgaggacagc gtgatcttca ccgacaagat catccgcagc 4441 aacgccaccg tggagcacct gcaccccatg ggcgataacg tgctggtggg cagcttcgcc 4501 cgcaccttca gcctgcgcga cggcggctac tacagcttcg tggtggacag ccacatgcac 4561 ttcaagagcg ccatccaccc cagcatcctg cagaacgggg gccccatgtt cgccttccgc 4621 cgcgtggagg agctgcacag caacaccgag ctgggcatcg tggagtacca gcacgccttc 4681 aagaccccca tcgccttcgc cagatcccgc gctcagtcgt ccaattctgc cgtggacggc 4741 accgccggac ccggctccac cggatctcgc gagggcagag gaagtcttct aacatgcggt 4801 gacgtggagg agaatcccgg ccctatgacc gagtacaagc ccacggtgcg cctcgccacc 4861 cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc 4921 acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct gcaagaactc 4981 ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg 5041 gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc 5101 ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggaaggcctc 5161 ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc 5221 gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag 5281 cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag 5341 cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc 5401 atgacccgca agcccggtgc ctgaaatcaa cctctggatt acaaaatttg tgaaagattg 5461 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct 5521 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg 5581 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact 5641 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc 5701 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc 5761 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa 5821 tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc 5881 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg 5941 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg 6001 gccgcctccc cgcctggtac ctttaagacc aatgacttac aaggcagctg tagatcttag 6061 ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac gaagataaga 6121 tctgcttttt gcttgtactg ggtctctctg gttagaccag atctgagcct gggagctctc 6181 tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt 6241 agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc 6301 agtgtggaaa atctctagca gtagtagttc atgtcatctt attattcagt atttataact 6361 tgcaaagaaa tgaatatcag agagtgagag gaacttgttt attgcagctt ataatggtta 6421 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag 6481 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggctctagc tatcccgccc 6541 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc 6601 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag 6661 aagtagtgag gaggcttttt tggaggccta gacttttgca gagaccaaat tcgtaatcat 6721 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc 6781 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc 6841 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat 6901 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac 6961 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt 7021 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca 7081 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc 7141 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact 7201 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct 7261 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag 7321 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca 7381 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa 7441 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc 7501 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag 7561 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg 7621 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca 7681 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc 7741 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag 7801 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata 7861 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat 7921 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg 7981 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc 8041 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc 8101 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc 8161 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc 8221 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc 8281 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa 8341 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat 8401 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata 8461 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca 8521 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag 8581 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc 8641 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc 8701 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata 8761 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta 8821 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta 8881 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg 8941 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt 9001 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg 9061 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt 9121 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg 9181 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct 9241 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg 9301 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctg //