LOCUS V030276 9166 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V030276 VERSION V030276 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..3192 /label="TNFAIP8L3(NM_207381)" /note="TNFAIP8L3(NM_207381)" /gene="TNFAIP8L3" CDS 3199..3264 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3308..3519 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3532..3800 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 3829..3838 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 3859..4521 /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 4591..4644 /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 4645..5244 /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 5245..5833 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5907..6140 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6212..6333 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6373..6508 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6541..6557) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6565..6581 /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(6589..6619) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6634..6655 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6943..7531) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7702..8562) /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(8563..8667) /gene="bla" /label="AmpR promoter" primer_bind 9141..9157 /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 ggaaaccacg gcaaaaccca 2341 agcacactgg tttccacact ctgtgaggca gagccaaagg ggaaactgtg ggtcaacgga 2401 tatgcaggga cccaaggcac aagggatgcc acgttacaaa caagactcat ccccttatct 2461 tttcatcttc agaggggaaa gggacttgcc gccccgctgt ccgccctgag tgcgccgcgg 2521 ctgcccgagc gccccgcaga cgggcgggtg gccgtggacg cccagccagc agcccgcagc 2581 atggattcgg attccgggga gcagagcgag ggcgagcccg tgaccgccgc aggtcctgat 2641 gtttttagtt caaagagtct tgcgcttcaa gcccagaaga agattctgag caaaatagcc 2701 agcaaaactg tggccaacat gttgattgat gacaccagca gcgagatctt tgatgagctc 2761 tacaaagtca ccaaagagca cacacacaac aagaaggaag cccacaagat catgaaagac 2821 ttaatcaagg tggcgatcaa aatcgggatc ctctaccgga acaaccagtt tagccaagag 2881 gagctggtta ttgtggagaa gttccggaag aagctgaacc agaccgccat gaccattgtc 2941 agcttctatg aggtggaata caccttcgat aggaacgtgc tctccaatct cctgcatgag 3001 tgcaaggacc tggtgcatga actggtgcag cggcacctga cgcccaggac ccacgggcgc 3061 atcaaccacg tctttaacca ctttgccgat gtggagttcc tctccaccct ctatagtctg 3121 gatggagact gtaggcccaa cctcaagagg atttgtgaag gaatcaataa gttgctagat 3181 gagaaagtcc ttggatccga ctacaaagac catgacggtg attataaaga tcatgacatc 3241 gactacaagg atgacgatga caagtagcgg ccgcgaagga tctgcgatcg ctccggtgcc 3301 cgtcagtggg cagagcgcac atcgcccaca gtccccgaga agttgggggg aggggtcggc 3361 aattgaacgg gtgcctagag aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac 3421 tggctccgcc tttttcccga gggtggggga gaaccgtata taagtgcagt agtcgccgtg 3481 aacgttcttt ttcgcaacgg gtttgccgcc agaacacagc tgaagcttcg aggggctcgc 3541 atctctcctt cacgcgcccg ccgccctacc tgaggccgcc atccacgccg gttgagtcgc 3601 gttctgccgc ctcccgcctg tggtgcctcc tgaactgcgt ccgccgtcta ggtaagttta 3661 aagctcaggt cgagaccggg cctttgtccg gcgctccctt ggagcctacc tagactcagc 3721 cggctctcca cgctttgcct gaccctgctt gctcaactct acgtctttgt ttcgttttct 3781 gttctgcgcc gttacagatc caagctgtga ccggcgccta cgctagacgc caccatggag 3841 agcgacgaga gcggcctgcc cgccatggag atcgagtgcc gcatcaccgg caccctgaac 3901 ggcgtggagt tcgagctggt gggcggcgga gagggcaccc ccaagcaggg ccgcatgacc 3961 aacaagatga agagcaccaa aggcgccctg accttcagcc cctacctgct gagccacgtg 4021 atgggctacg gcttctacca cttcggcacc taccccagcg gctacgagaa ccccttcctg 4081 cacgccatca acaacggcgg ctacaccaac acccgcatcg agaagtacga ggacggcggc 4141 gtgctgcacg tgagcttcag ctaccgctac gaggccggcc gcgtgatcgg cgacttcaag 4201 gtggtgggca ccggcttccc cgaggacagc gtgatcttca ccgacaagat catccgcagc 4261 aacgccaccg tggagcacct gcaccccatg ggcgataacg tgctggtggg cagcttcgcc 4321 cgcaccttca gcctgcgcga cggcggctac tacagcttcg tggtggacag ccacatgcac 4381 ttcaagagcg ccatccaccc cagcatcctg cagaacgggg gccccatgtt cgccttccgc 4441 cgcgtggagg agctgcacag caacaccgag ctgggcatcg tggagtacca gcacgccttc 4501 aagaccccca tcgccttcgc cagatcccgc gctcagtcgt ccaattctgc cgtggacggc 4561 accgccggac ccggctccac cggatctcgc gagggcagag gaagtcttct aacatgcggt 4621 gacgtggagg agaatcccgg ccctatgacc gagtacaagc ccacggtgcg cctcgccacc 4681 cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc 4741 acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct gcaagaactc 4801 ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg 4861 gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc 4921 ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggaaggcctc 4981 ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc 5041 gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag 5101 cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag 5161 cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc 5221 atgacccgca agcccggtgc ctgaaatcaa cctctggatt acaaaatttg tgaaagattg 5281 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct 5341 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg 5401 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact 5461 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc 5521 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc 5581 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa 5641 tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc 5701 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg 5761 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg 5821 gccgcctccc cgcctggtac ctttaagacc aatgacttac aaggcagctg tagatcttag 5881 ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac gaagataaga 5941 tctgcttttt gcttgtactg ggtctctctg gttagaccag atctgagcct gggagctctc 6001 tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt 6061 agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc 6121 agtgtggaaa atctctagca gtagtagttc atgtcatctt attattcagt atttataact 6181 tgcaaagaaa tgaatatcag agagtgagag gaacttgttt attgcagctt ataatggtta 6241 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag 6301 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggctctagc tatcccgccc 6361 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc 6421 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag 6481 aagtagtgag gaggcttttt tggaggccta gacttttgca gagaccaaat tcgtaatcat 6541 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc 6601 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc 6661 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat 6721 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac 6781 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt 6841 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca 6901 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc 6961 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact 7021 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct 7081 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag 7141 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca 7201 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa 7261 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc 7321 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag 7381 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg 7441 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca 7501 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc 7561 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag 7621 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata 7681 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat 7741 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg 7801 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc 7861 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc 7921 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc 7981 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc 8041 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc 8101 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa 8161 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat 8221 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata 8281 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca 8341 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag 8401 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc 8461 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc 8521 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata 8581 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta 8641 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta 8701 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg 8761 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt 8821 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg 8881 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt 8941 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg 9001 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct 9061 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg 9121 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctg //