LOCUS V035550 10027 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V035550 VERSION V035550 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..4053 /label="YTHDF2(NM_016258)" /note="YTHDF2(NM_016258)" /gene="YTHDF2" CDS 4060..4125 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 4169..4380 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4393..4661 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4690..4699 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4720..5382 /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 5452..5505 /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 5506..6105 /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 6106..6694 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6768..7001 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 7073..7194 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 7234..7369 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(7402..7418) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7426..7442 /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(7450..7480) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7495..7516 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7804..8392) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8563..9423) /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(9424..9528) /gene="bla" /label="AmpR promoter" primer_bind 10002..10018 /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 gccaccatgt cggccagcag cctcttggag 2341 cagagaccaa aaggtcaagg aaacaaagta caaaatggat ctgtacatca aaaggatgga 2401 ttaaacgatg atgattttga accttacttg agtccacagg caaggcccaa taatgcatat 2461 actgccatgt cagattccta cttacccagt tactacagtc cctccattgg cttctcctat 2521 tctttgggtg aagctgcttg gtctacgggg ggtgacacag ccatgcccta cttaacttct 2581 tatggacagc tgagcaacgg agagccccac ttcctaccag atgcaatgtt tgggcaacca 2641 ggagccctag gtagcactcc atttcttggt cagcatggtt ttaatttctt tcccagtggg 2701 attgacttct cagcatgggg aaataacagt tctcagggac agtctactca gagctctgga 2761 tatagtagca attatgctta tgcacctagc tccttaggtg gagccatgat tgatggacag 2821 tcagcttttg ccaatgagac cctcaataag gctcctggca tgaatactat agaccaaggg 2881 atggcagcac tgaagttggg tagcacagaa gttgcaagca atgttccaaa agttgtaggt 2941 tctgctgttg gtagcgggtc cattactagt aacatcgtgg cttccaatag tttgcctcca 3001 gccaccattg ctcctccaaa accagcatct tgggctgata ttgctagcaa gcctgcaaaa 3061 cagcaaccta aactgaagac caagaatggc attgcagggt caagtcttcc gccacccccg 3121 ataaagcata acatggatat tggaacttgg gataacaagg gtcccgttgc aaaagccccc 3181 tcacaggctt tggttcagaa tataggtcag ccaacccagg ggtctcctca gcctgtaggt 3241 cagcaggcta acaatagccc accagtggct caggcatcag tagggcaaca gacacagcca 3301 ttgcctccac ctccaccaca gcctgcccag ctttcagtcc agcaacaggc agctcagcca 3361 acccgctggg tagcacctcg gaaccgtggc agtgggttcg gtcataatgg ggtggatggt 3421 aatggagtag gacagtctca ggctggttct ggatctactc cttcagaacc ccacccagtg 3481 ttggagaagc ttcggtccat taataactat aaccccaaag attttgactg gaatctgaaa 3541 catggccggg ttttcatcat taagagctac tctgaggacg atattcaccg ttccattaag 3601 tataatattt ggtgcagcac agagcatggt aacaagagac tggatgctgc ttatcgttcc 3661 atgaacggga aaggccccgt ttacttactt ttcagtgtca acggcagtgg acacttctgt 3721 ggcgtggcag aaatgaaatc tgctgtggac tacaacacat gtgcaggtgt gtggtcccag 3781 gacaaatgga agggtcgttt tgatgtcagg tggatttttg tgaaggacgt tcccaatagc 3841 caactgcgac acattcgcct agagaacaac gagaataaac cagtgaccaa ctctagggac 3901 actcaggaag tgcctctgga aaaggctaag caggtgttga aaattatagc cagctacaag 3961 cacaccactt ccatttttga tgacttctca cactatgaga aacgccaaga ggaagaagaa 4021 agtgttaaaa aggaacgtca aggtcgtggg aaaggatccg actacaaaga ccatgacggt 4081 gattataaag atcatgacat cgactacaag gatgacgatg acaagtagcg gccgcgaagg 4141 atctgcgatc gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag 4201 aagttggggg gaggggtcgg caattgaacg ggtgcctaga gaaggtggcg cggggtaaac 4261 tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat 4321 ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag 4381 ctgaagcttc gaggggctcg catctctcct tcacgcgccc gccgccctac ctgaggccgc 4441 catccacgcc ggttgagtcg cgttctgccg cctcccgcct gtggtgcctc ctgaactgcg 4501 tccgccgtct aggtaagttt aaagctcagg tcgagaccgg gcctttgtcc ggcgctccct 4561 tggagcctac ctagactcag ccggctctcc acgctttgcc tgaccctgct tgctcaactc 4621 tacgtctttg tttcgttttc tgttctgcgc cgttacagat ccaagctgtg accggcgcct 4681 acgctagacg ccaccatgga gagcgacgag agcggcctgc ccgccatgga gatcgagtgc 4741 cgcatcaccg gcaccctgaa cggcgtggag ttcgagctgg tgggcggcgg agagggcacc 4801 cccaagcagg gccgcatgac caacaagatg aagagcacca aaggcgccct gaccttcagc 4861 ccctacctgc tgagccacgt gatgggctac ggcttctacc acttcggcac ctaccccagc 4921 ggctacgaga accccttcct gcacgccatc aacaacggcg gctacaccaa cacccgcatc 4981 gagaagtacg aggacggcgg cgtgctgcac gtgagcttca gctaccgcta cgaggccggc 5041 cgcgtgatcg gcgacttcaa ggtggtgggc accggcttcc ccgaggacag cgtgatcttc 5101 accgacaaga tcatccgcag caacgccacc gtggagcacc tgcaccccat gggcgataac 5161 gtgctggtgg gcagcttcgc ccgcaccttc agcctgcgcg acggcggcta ctacagcttc 5221 gtggtggaca gccacatgca cttcaagagc gccatccacc ccagcatcct gcagaacggg 5281 ggccccatgt tcgccttccg ccgcgtggag gagctgcaca gcaacaccga gctgggcatc 5341 gtggagtacc agcacgcctt caagaccccc atcgccttcg ccagatcccg cgctcagtcg 5401 tccaattctg ccgtggacgg caccgccgga cccggctcca ccggatctcg cgagggcaga 5461 ggaagtcttc taacatgcgg tgacgtggag gagaatcccg gccctatgac cgagtacaag 5521 cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc 5581 gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg 5641 gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg 5701 gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg 5761 gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg 5821 cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg 5881 gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc 5941 cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc 6001 cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc 6061 gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgaaatca acctctggat 6121 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt 6181 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc 6241 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg 6301 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc 6361 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa 6421 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat 6481 tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc 6541 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt 6601 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag 6661 acgagtcgga tctccctttg ggccgcctcc ccgcctggta cctttaagac caatgactta 6721 caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat 6781 tcactcccaa cgaagataag atctgctttt tgcttgtact gggtctctct ggttagacca 6841 gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag 6901 cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag 6961 atccctcaga cccttttagt cagtgtggaa aatctctagc agtagtagtt catgtcatct 7021 tattattcag tatttataac ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt 7081 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc 7141 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt 7201 ctggctctag ctatcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc 7261 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc 7321 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct agacttttgc 7381 agagaccaaa ttcgtaatca tgtcatagct gtttcctgtg tgaaattgtt atccgctcac 7441 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt 7501 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc 7561 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg 7621 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt 7681 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa 7741 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc 7801 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag 7861 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt 7921 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg 7981 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg 8041 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg 8101 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac 8161 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg 8221 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt 8281 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg 8341 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc 8401 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt 8461 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt 8521 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag 8581 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt 8641 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc 8701 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc 8761 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg 8821 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac 8881 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg 8941 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc 9001 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact 9061 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc 9121 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat 9181 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc 9241 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac 9301 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa 9361 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact 9421 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg 9481 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg 9541 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag 9601 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca 9661 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc 9721 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc 9781 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag 9841 gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg 9901 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg 9961 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtgc 10021 caagctg //