LOCUS V025862 10087 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V025862 VERSION V025862 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..4113 /label="ZC3H12A(NM_001323550)" /note="ZC3H12A(NM_001323550)" /gene="ZC3H12A" CDS 4120..4185 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 4229..4440 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4453..4721 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4750..4759 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4780..5442 /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 5512..5565 /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 5566..6165 /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 6166..6754 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6828..7061 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 7133..7254 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 7294..7429 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(7462..7478) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7486..7502 /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(7510..7540) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7555..7576 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7864..8452) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8623..9483) /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(9484..9588) /gene="bla" /label="AmpR promoter" primer_bind 10062..10078 /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 gtggcccctg tggagagaag 2341 cctgtcctgg aagccagccc caccatgagt ctgtgggaat ttgaggacag ccacagccgt 2401 cagggcaccc caaggccggg tcaagagctg gccgctgagg aggcctcggc cctggaactg 2461 cagatgaagg tggacttctt ccggaagctg ggctattcat ccacggagat ccacagcgtc 2521 ctgcagaagc tgggcgtcca ggcagacacc aacacggtgc tgggtgagct ggtgaaacac 2581 gggacagcca ccgagcggga gcgccagacc tcaccggacc cctgccctca gctccctcta 2641 gtcccgcggg gtggtggcac ccctaaggct cccaacctgg agcctccact cccagaagag 2701 gaaaaggagg gcagcgacct gagaccagtg gtcatcgatg ggagcaacgt ggccatgagc 2761 catgggaaca aggaggtctt ctcctgccgg ggcatcctgc tggcagtgaa ctggtttctg 2821 gagcggggcc acacagacat cacagtgttt gtgccatcct ggaggaagga gcagcctcgg 2881 cccgacgtgc ccatcacaga ccagcacatc ctgcgggaac tggagaagaa gaagatcctg 2941 gtgttcacac catcacgacg cgtgggtggc aagcgggtgg tgtgctatga cgacagattc 3001 attgtgaagc tggcctacga gtctgacggg atcgtggttt ccaacgacac ataccgtgac 3061 ctccaaggcg agcggcagga gtggaagcgc ttcatcgagg agcggctgct catgtactcc 3121 ttcgtcaatg acaagtttat gccccctgat gacccactgg gccggcacgg gcccagcctg 3181 gacaacttcc tgcgtaagaa gccactcact ttggagcaca ggaagcagcc gtgtccctat 3241 ggaaggaaat gcacctatgg gatcaagtgc cgattcttcc acccagagcg gccaagctgc 3301 ccccagcgct ctgtggcaga tgagctccgt gccaatgctc tcctctcacc ccccagagcc 3361 ccaagcaagg acaaaaatgg ccggcggcct tcaccttcat cccagtccag ctctctgcta 3421 acagagagtg agcagtgcag cctggatggg aagaagctgg gggcccaggc atccccaggg 3481 tcccgccaag agggtctaac acagacctat gccccatcag gcaggagcct cgcacctagc 3541 gggggcagtg gcagcagctt tgggcccaca gactggctcc cacagacgct ggactcactc 3601 ccgtacgtct cccaggattg cctggactcg ggcattggct ccctggagag ccagatgtcg 3661 gaactttggg gggttcgagg aggaggccct ggtgagccgg gcccaccccg agccccttac 3721 acgggctaca gtccctatgg atctgagctc ccagccaccg cagccttctc tgcctttggc 3781 cgggccatgg gtgctggcca cttcagtgtc cctgccgact acccacccgc gccccctgcc 3841 tttccacctc gagagtactg gtctgaacca tacccactgc ccccacccac atcagtcctt 3901 caggagcccc cagtgcagag cccaggggct ggcaggagcc cgtggggcag ggcaggcagc 3961 ctggccaagg agcaggccag cgtgtatact aagctgtgtg gtgtgtttcc cccgcacctg 4021 gtggaggctg tgatggggcg cttcccacag ctcctggacc cccagcagct ggctgccgag 4081 atcctctcct acaagtccca gcaccccagt gagggatccg actacaaaga ccatgacggt 4141 gattataaag atcatgacat cgactacaag gatgacgatg acaagtagcg gccgcgaagg 4201 atctgcgatc gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag 4261 aagttggggg gaggggtcgg caattgaacg ggtgcctaga gaaggtggcg cggggtaaac 4321 tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat 4381 ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag 4441 ctgaagcttc gaggggctcg catctctcct tcacgcgccc gccgccctac ctgaggccgc 4501 catccacgcc ggttgagtcg cgttctgccg cctcccgcct gtggtgcctc ctgaactgcg 4561 tccgccgtct aggtaagttt aaagctcagg tcgagaccgg gcctttgtcc ggcgctccct 4621 tggagcctac ctagactcag ccggctctcc acgctttgcc tgaccctgct tgctcaactc 4681 tacgtctttg tttcgttttc tgttctgcgc cgttacagat ccaagctgtg accggcgcct 4741 acgctagacg ccaccatgga gagcgacgag agcggcctgc ccgccatgga gatcgagtgc 4801 cgcatcaccg gcaccctgaa cggcgtggag ttcgagctgg tgggcggcgg agagggcacc 4861 cccaagcagg gccgcatgac caacaagatg aagagcacca aaggcgccct gaccttcagc 4921 ccctacctgc tgagccacgt gatgggctac ggcttctacc acttcggcac ctaccccagc 4981 ggctacgaga accccttcct gcacgccatc aacaacggcg gctacaccaa cacccgcatc 5041 gagaagtacg aggacggcgg cgtgctgcac gtgagcttca gctaccgcta cgaggccggc 5101 cgcgtgatcg gcgacttcaa ggtggtgggc accggcttcc ccgaggacag cgtgatcttc 5161 accgacaaga tcatccgcag caacgccacc gtggagcacc tgcaccccat gggcgataac 5221 gtgctggtgg gcagcttcgc ccgcaccttc agcctgcgcg acggcggcta ctacagcttc 5281 gtggtggaca gccacatgca cttcaagagc gccatccacc ccagcatcct gcagaacggg 5341 ggccccatgt tcgccttccg ccgcgtggag gagctgcaca gcaacaccga gctgggcatc 5401 gtggagtacc agcacgcctt caagaccccc atcgccttcg ccagatcccg cgctcagtcg 5461 tccaattctg ccgtggacgg caccgccgga cccggctcca ccggatctcg cgagggcaga 5521 ggaagtcttc taacatgcgg tgacgtggag gagaatcccg gccctatgac cgagtacaag 5581 cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc 5641 gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg 5701 gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg 5761 gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg 5821 gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg 5881 cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg 5941 gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc 6001 cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc 6061 cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc 6121 gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgaaatca acctctggat 6181 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt 6241 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc 6301 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg 6361 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc 6421 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa 6481 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat 6541 tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc 6601 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt 6661 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag 6721 acgagtcgga tctccctttg ggccgcctcc ccgcctggta cctttaagac caatgactta 6781 caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat 6841 tcactcccaa cgaagataag atctgctttt tgcttgtact gggtctctct ggttagacca 6901 gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag 6961 cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag 7021 atccctcaga cccttttagt cagtgtggaa aatctctagc agtagtagtt catgtcatct 7081 tattattcag tatttataac ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt 7141 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc 7201 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt 7261 ctggctctag ctatcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc 7321 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc 7381 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct agacttttgc 7441 agagaccaaa ttcgtaatca tgtcatagct gtttcctgtg tgaaattgtt atccgctcac 7501 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt 7561 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc 7621 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg 7681 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt 7741 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa 7801 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc 7861 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag 7921 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt 7981 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg 8041 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg 8101 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg 8161 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac 8221 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg 8281 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt 8341 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg 8401 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc 8461 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt 8521 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt 8581 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag 8641 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt 8701 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc 8761 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc 8821 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg 8881 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac 8941 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg 9001 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc 9061 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact 9121 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc 9181 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat 9241 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc 9301 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac 9361 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa 9421 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact 9481 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg 9541 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg 9601 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag 9661 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca 9721 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc 9781 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc 9841 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag 9901 gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg 9961 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg 10021 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtgc 10081 caagctg //