LOCUS V033515 9406 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V033515 VERSION V033515 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..3432 /label="TRIB1(NM_025195)" /note="TRIB1(NM_025195)" /gene="TRIB1" CDS 3439..3504 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3548..3759 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3772..4040 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4069..4078 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4099..4761 /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 4831..4884 /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 4885..5484 /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 5485..6073 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6147..6380 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6452..6573 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6613..6748 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6781..6797) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6805..6821 /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(6829..6859) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6874..6895 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7183..7771) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7942..8802) /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(8803..8907) /gene="bla" /label="AmpR promoter" primer_bind 9381..9397 /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 gccaccatgc gggtcggtcc ggtgcgctct 2341 gccatgagcg gcgcctcgca gccccgcggc ccggccctgc tcttcccagc cacccgaggc 2401 gtcccggcca aacgcctgct ggacgccgac gacgcggcgg ctgtggcggc caagtgcccg 2461 cgcctctccg agtgctccag ccccccggac tacctcagcc cccccggctc gccctgcagc 2521 ccgcagcccc cgcctgccgc tccgggggcc ggcggaggct ccgggagcgc gccggggccc 2581 agccgcatcg ccgactacct gctgctgccc ctagccgagc gcgagcatgt gtcccgggcg 2641 ctgtgcatcc acactggacg cgagctgcgc tgcaaggtgt ttcccattaa acactaccag 2701 gacaaaatca ggccttacat ccagctgcca tcgcacagca acattactgg cattgtggaa 2761 gtgatccttg gggaaaccaa ggcctatgtc ttctttgaga aggactttgg ggacatgcac 2821 tcctatgtgc gaagccggaa gaggctgcgg gaagaggaag ccgcccggct cttcaagcag 2881 attgtctccg ccgtcgccca ctgccaccag tcagccatcg tgctggggga cctgaagctt 2941 aggaagttcg tcttctccac ggaggagaga acccagctta gactagaaag tctagaagac 3001 acacacataa tgaaggggga agatgatgct ttgtcagaca aacatggctg cccagcctac 3061 gtgagccctg agatcctcaa caccactggg acctactccg gaaaggctgc ggacgtttgg 3121 agcctggggg tgatgctcta cacccttctg gttggacgat accccttcca tgactcagac 3181 cccagtgccc ttttctccaa aattcggcgt ggacagttct gcattcctga gcacatttcc 3241 cccaaagcca ggtgcctcat tcgcagcctc ttgagacggg agccctccga gagactcact 3301 gcccccgaga tcctactgca cccctggttt gagtccgtct tggaacccgg gtacatcgac 3361 tcagaaatag gaacttcaga ccagattgtt ccagagtacc aggaggacag tgacattagt 3421 tccttcttct gcggatccga ctacaaagac catgacggtg attataaaga tcatgacatc 3481 gactacaagg atgacgatga caagtagcgg ccgcgaagga tctgcgatcg ctccggtgcc 3541 cgtcagtggg cagagcgcac atcgcccaca gtccccgaga agttgggggg aggggtcggc 3601 aattgaacgg gtgcctagag aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac 3661 tggctccgcc tttttcccga gggtggggga gaaccgtata taagtgcagt agtcgccgtg 3721 aacgttcttt ttcgcaacgg gtttgccgcc agaacacagc tgaagcttcg aggggctcgc 3781 atctctcctt cacgcgcccg ccgccctacc tgaggccgcc atccacgccg gttgagtcgc 3841 gttctgccgc ctcccgcctg tggtgcctcc tgaactgcgt ccgccgtcta ggtaagttta 3901 aagctcaggt cgagaccggg cctttgtccg gcgctccctt ggagcctacc tagactcagc 3961 cggctctcca cgctttgcct gaccctgctt gctcaactct acgtctttgt ttcgttttct 4021 gttctgcgcc gttacagatc caagctgtga ccggcgccta cgctagacgc caccatggag 4081 agcgacgaga gcggcctgcc cgccatggag atcgagtgcc gcatcaccgg caccctgaac 4141 ggcgtggagt tcgagctggt gggcggcgga gagggcaccc ccaagcaggg ccgcatgacc 4201 aacaagatga agagcaccaa aggcgccctg accttcagcc cctacctgct gagccacgtg 4261 atgggctacg gcttctacca cttcggcacc taccccagcg gctacgagaa ccccttcctg 4321 cacgccatca acaacggcgg ctacaccaac acccgcatcg agaagtacga ggacggcggc 4381 gtgctgcacg tgagcttcag ctaccgctac gaggccggcc gcgtgatcgg cgacttcaag 4441 gtggtgggca ccggcttccc cgaggacagc gtgatcttca ccgacaagat catccgcagc 4501 aacgccaccg tggagcacct gcaccccatg ggcgataacg tgctggtggg cagcttcgcc 4561 cgcaccttca gcctgcgcga cggcggctac tacagcttcg tggtggacag ccacatgcac 4621 ttcaagagcg ccatccaccc cagcatcctg cagaacgggg gccccatgtt cgccttccgc 4681 cgcgtggagg agctgcacag caacaccgag ctgggcatcg tggagtacca gcacgccttc 4741 aagaccccca tcgccttcgc cagatcccgc gctcagtcgt ccaattctgc cgtggacggc 4801 accgccggac ccggctccac cggatctcgc gagggcagag gaagtcttct aacatgcggt 4861 gacgtggagg agaatcccgg ccctatgacc gagtacaagc ccacggtgcg cctcgccacc 4921 cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc 4981 acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct gcaagaactc 5041 ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg 5101 gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc 5161 ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggaaggcctc 5221 ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc 5281 gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag 5341 cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag 5401 cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc 5461 atgacccgca agcccggtgc ctgaaatcaa cctctggatt acaaaatttg tgaaagattg 5521 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct 5581 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg 5641 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact 5701 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc 5761 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc 5821 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa 5881 tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc 5941 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg 6001 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg 6061 gccgcctccc cgcctggtac ctttaagacc aatgacttac aaggcagctg tagatcttag 6121 ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac gaagataaga 6181 tctgcttttt gcttgtactg ggtctctctg gttagaccag atctgagcct gggagctctc 6241 tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt 6301 agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc 6361 agtgtggaaa atctctagca gtagtagttc atgtcatctt attattcagt atttataact 6421 tgcaaagaaa tgaatatcag agagtgagag gaacttgttt attgcagctt ataatggtta 6481 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag 6541 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggctctagc tatcccgccc 6601 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc 6661 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag 6721 aagtagtgag gaggcttttt tggaggccta gacttttgca gagaccaaat tcgtaatcat 6781 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc 6841 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc 6901 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat 6961 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac 7021 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt 7081 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca 7141 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc 7201 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact 7261 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct 7321 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag 7381 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca 7441 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa 7501 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc 7561 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag 7621 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg 7681 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca 7741 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc 7801 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag 7861 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata 7921 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat 7981 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg 8041 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc 8101 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc 8161 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc 8221 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc 8281 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc 8341 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa 8401 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat 8461 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata 8521 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca 8581 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag 8641 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc 8701 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc 8761 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata 8821 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta 8881 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta 8941 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg 9001 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt 9061 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg 9121 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt 9181 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg 9241 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct 9301 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg 9361 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctg //