LOCUS V041169 9547 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V041169 VERSION V041169 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..3573 /label="Sox17(NM_011441)" /note="Sox17(NM_011441)" /gene="Sox17" CDS 3580..3645 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3689..3900 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3913..4181 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4210..4219 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4240..4902 /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 4972..5025 /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 5026..5625 /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 5626..6214 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6288..6521 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6593..6714 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6754..6889 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6922..6938) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6946..6962 /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(6970..7000) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7015..7036 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7324..7912) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8083..8943) /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(8944..9048) /gene="bla" /label="AmpR promoter" primer_bind 9522..9538 /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 gcagcccgga tgcgggatac 2341 gccagtgacg accagagcca gccccggagc gcgcagcccg cggtgatggc agggttgggc 2401 ccctgtccct gggccgagtc cctgagcccc ctcggggatg taaaggtgaa aggcgaggtg 2461 gtggcgagta gcggggcgcc agccgggacg tcgggccgag ccaaagcgga gtctcgcatc 2521 cggcggccga tgaacgcctt tatggtgtgg gccaaagacg aacgcaagcg gttggcacag 2581 cagaacccag atctgcacaa cgcagagcta agcaagatgc taggcaagtc ttggaaggcg 2641 ttgaccttgg cagagaagcg gcccttcgtg gaagaggccg agcggctgcg cgtgcagcat 2701 atgcaggacc accccaacta caagtaccgg ccgcggcggc gcaagcaggt gaagcgcatg 2761 aagcgggtgg agggaggctt cctgcacgct ctcgtcgagc cccaggccgg cgcgcttggt 2821 cccgagggcg gccgcgtggc catggatggc ctgggtctgc ctttcccgga gccgggctat 2881 ccggccggtc ctccgctgat gtctccgcac atgggccccc actatcggga ctgccaggga 2941 ctgggcgctc ccgcgctcga cggctaccct ctgcccactc cggacacatc cccgctggat 3001 ggcgtggagc aggacccggc tttctttgca gccccgctgc caggggactg cccggcggcc 3061 ggcacctaca cttacgctcc agtctcggac tatgcagtgt ccgtagagcc gcccgctggc 3121 cccatgcgag tggggccgga cccctcgggc cctgcgatgc cggggatcct ggcgcccccc 3181 agcgctctgc acctgtacta cggcgcgatg ggctcgcccg ccgcaagtgc ggggcgcggt 3241 ttccacgcgc aaccccagca gccgctgcaa ccgcaggcac cgccgccgcc accgcagcag 3301 cagcacccag cgcacggccc cgggcaacct tcgccccctc ccgaggctct gccctgccgg 3361 gatggcacgg aatccaacca gcccactgag ctcctagggg aggtggaccg cacggaattc 3421 gaacagtatc tgccctttgt gtataagccc gagatgggtc ttccctacca gggacacgac 3481 tgcggagtga acctctcaga cagccacgga gccatttcct ccgtggtgtc cgacgctagc 3541 tcagcggtct actattgcaa ctaccccgac attggatccg actacaaaga ccatgacggt 3601 gattataaag atcatgacat cgactacaag gatgacgatg acaagtagcg gccgcgaagg 3661 atctgcgatc gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag 3721 aagttggggg gaggggtcgg caattgaacg ggtgcctaga gaaggtggcg cggggtaaac 3781 tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat 3841 ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag 3901 ctgaagcttc gaggggctcg catctctcct tcacgcgccc gccgccctac ctgaggccgc 3961 catccacgcc ggttgagtcg cgttctgccg cctcccgcct gtggtgcctc ctgaactgcg 4021 tccgccgtct aggtaagttt aaagctcagg tcgagaccgg gcctttgtcc ggcgctccct 4081 tggagcctac ctagactcag ccggctctcc acgctttgcc tgaccctgct tgctcaactc 4141 tacgtctttg tttcgttttc tgttctgcgc cgttacagat ccaagctgtg accggcgcct 4201 acgctagacg ccaccatgga gagcgacgag agcggcctgc ccgccatgga gatcgagtgc 4261 cgcatcaccg gcaccctgaa cggcgtggag ttcgagctgg tgggcggcgg agagggcacc 4321 cccaagcagg gccgcatgac caacaagatg aagagcacca aaggcgccct gaccttcagc 4381 ccctacctgc tgagccacgt gatgggctac ggcttctacc acttcggcac ctaccccagc 4441 ggctacgaga accccttcct gcacgccatc aacaacggcg gctacaccaa cacccgcatc 4501 gagaagtacg aggacggcgg cgtgctgcac gtgagcttca gctaccgcta cgaggccggc 4561 cgcgtgatcg gcgacttcaa ggtggtgggc accggcttcc ccgaggacag cgtgatcttc 4621 accgacaaga tcatccgcag caacgccacc gtggagcacc tgcaccccat gggcgataac 4681 gtgctggtgg gcagcttcgc ccgcaccttc agcctgcgcg acggcggcta ctacagcttc 4741 gtggtggaca gccacatgca cttcaagagc gccatccacc ccagcatcct gcagaacggg 4801 ggccccatgt tcgccttccg ccgcgtggag gagctgcaca gcaacaccga gctgggcatc 4861 gtggagtacc agcacgcctt caagaccccc atcgccttcg ccagatcccg cgctcagtcg 4921 tccaattctg ccgtggacgg caccgccgga cccggctcca ccggatctcg cgagggcaga 4981 ggaagtcttc taacatgcgg tgacgtggag gagaatcccg gccctatgac cgagtacaag 5041 cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc 5101 gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg 5161 gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg 5221 gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg 5281 gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg 5341 cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg 5401 gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc 5461 cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc 5521 cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc 5581 gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgaaatca acctctggat 5641 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt 5701 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc 5761 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg 5821 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc 5881 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa 5941 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat 6001 tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc 6061 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt 6121 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag 6181 acgagtcgga tctccctttg ggccgcctcc ccgcctggta cctttaagac caatgactta 6241 caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat 6301 tcactcccaa cgaagataag atctgctttt tgcttgtact gggtctctct ggttagacca 6361 gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag 6421 cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag 6481 atccctcaga cccttttagt cagtgtggaa aatctctagc agtagtagtt catgtcatct 6541 tattattcag tatttataac ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt 6601 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc 6661 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt 6721 ctggctctag ctatcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc 6781 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc 6841 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct agacttttgc 6901 agagaccaaa ttcgtaatca tgtcatagct gtttcctgtg tgaaattgtt atccgctcac 6961 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt 7021 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc 7081 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg 7141 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt 7201 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa 7261 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc 7321 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag 7381 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt 7441 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg 7501 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg 7561 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg 7621 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac 7681 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg 7741 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt 7801 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg 7861 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc 7921 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt 7981 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt 8041 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag 8101 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt 8161 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc 8221 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc 8281 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg 8341 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac 8401 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg 8461 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc 8521 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact 8581 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc 8641 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat 8701 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc 8761 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac 8821 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa 8881 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact 8941 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg 9001 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg 9061 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag 9121 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca 9181 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc 9241 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc 9301 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag 9361 gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg 9421 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg 9481 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtgc 9541 caagctg //