LOCUS V031662 9007 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V031662 VERSION V031662 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..3033 /label="FGF3(NM_005247)" /note="FGF3(NM_005247)" /gene="FGF3" CDS 3040..3105 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3149..3360 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3373..3641 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 3670..3679 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 3700..4362 /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 4432..4485 /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 4486..5085 /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 5086..5674 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5748..5981 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6053..6174 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6214..6349 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6382..6398) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6406..6422 /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(6430..6460) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6475..6496 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6784..7372) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7543..8403) /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(8404..8508) /gene="bla" /label="AmpR promoter" primer_bind 8982..8998 /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 gcctaatctg gctgctactg 2341 ctcagcctgc tggagcccgg ctggcccgca gcgggccctg gggcgcggtt gcggcgcgat 2401 gcgggcggcc gtggcggcgt ctacgagcac cttggcgggg cgccccggcg ccgcaagctc 2461 tactgcgcca cgaagtacca cctccagctg cacccgagcg gccgcgtcaa cggcagcctg 2521 gagaacagcg cctacagtat tttggagata acggcagtgg aggtgggcat tgtggccatc 2581 aggggtctct tctccgggcg gtacctggcc atgaacaaga ggggacgact ctatgcttcg 2641 gagcactaca gcgccgagtg cgagtttgtg gagcggatcc acgagctggg ctataatacg 2701 tatgcctccc ggctgtaccg gacggtgtct agtacgcctg gggcccgccg gcagcccagc 2761 gccgagagac tgtggtacgt gtctgtgaac ggcaagggcc ggccccgcag gggcttcaag 2821 acccgccgca cacagaagtc ctccctgttc ctgccccgcg tgctggacca cagggaccac 2881 gagatggtgc ggcagctaca gagtgggctg cccagacccc ctggtaaggg ggtccagccc 2941 cgacggcggc ggcagaagca gagcccggat aacctggagc cctctcacgt tcaggcttcg 3001 agactgggct cccagctgga ggccagtgcg cacggatccg actacaaaga ccatgacggt 3061 gattataaag atcatgacat cgactacaag gatgacgatg acaagtagcg gccgcgaagg 3121 atctgcgatc gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag 3181 aagttggggg gaggggtcgg caattgaacg ggtgcctaga gaaggtggcg cggggtaaac 3241 tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat 3301 ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag 3361 ctgaagcttc gaggggctcg catctctcct tcacgcgccc gccgccctac ctgaggccgc 3421 catccacgcc ggttgagtcg cgttctgccg cctcccgcct gtggtgcctc ctgaactgcg 3481 tccgccgtct aggtaagttt aaagctcagg tcgagaccgg gcctttgtcc ggcgctccct 3541 tggagcctac ctagactcag ccggctctcc acgctttgcc tgaccctgct tgctcaactc 3601 tacgtctttg tttcgttttc tgttctgcgc cgttacagat ccaagctgtg accggcgcct 3661 acgctagacg ccaccatgga gagcgacgag agcggcctgc ccgccatgga gatcgagtgc 3721 cgcatcaccg gcaccctgaa cggcgtggag ttcgagctgg tgggcggcgg agagggcacc 3781 cccaagcagg gccgcatgac caacaagatg aagagcacca aaggcgccct gaccttcagc 3841 ccctacctgc tgagccacgt gatgggctac ggcttctacc acttcggcac ctaccccagc 3901 ggctacgaga accccttcct gcacgccatc aacaacggcg gctacaccaa cacccgcatc 3961 gagaagtacg aggacggcgg cgtgctgcac gtgagcttca gctaccgcta cgaggccggc 4021 cgcgtgatcg gcgacttcaa ggtggtgggc accggcttcc ccgaggacag cgtgatcttc 4081 accgacaaga tcatccgcag caacgccacc gtggagcacc tgcaccccat gggcgataac 4141 gtgctggtgg gcagcttcgc ccgcaccttc agcctgcgcg acggcggcta ctacagcttc 4201 gtggtggaca gccacatgca cttcaagagc gccatccacc ccagcatcct gcagaacggg 4261 ggccccatgt tcgccttccg ccgcgtggag gagctgcaca gcaacaccga gctgggcatc 4321 gtggagtacc agcacgcctt caagaccccc atcgccttcg ccagatcccg cgctcagtcg 4381 tccaattctg ccgtggacgg caccgccgga cccggctcca ccggatctcg cgagggcaga 4441 ggaagtcttc taacatgcgg tgacgtggag gagaatcccg gccctatgac cgagtacaag 4501 cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc 4561 gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg 4621 gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg 4681 gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg 4741 gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg 4801 cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg 4861 gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc 4921 cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc 4981 cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc 5041 gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgaaatca acctctggat 5101 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt 5161 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc 5221 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg 5281 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc 5341 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa 5401 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat 5461 tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc 5521 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt 5581 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag 5641 acgagtcgga tctccctttg ggccgcctcc ccgcctggta cctttaagac caatgactta 5701 caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat 5761 tcactcccaa cgaagataag atctgctttt tgcttgtact gggtctctct ggttagacca 5821 gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag 5881 cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag 5941 atccctcaga cccttttagt cagtgtggaa aatctctagc agtagtagtt catgtcatct 6001 tattattcag tatttataac ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt 6061 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc 6121 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt 6181 ctggctctag ctatcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc 6241 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc 6301 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct agacttttgc 6361 agagaccaaa ttcgtaatca tgtcatagct gtttcctgtg tgaaattgtt atccgctcac 6421 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt 6481 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc 6541 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg 6601 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt 6661 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa 6721 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc 6781 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag 6841 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt 6901 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg 6961 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg 7021 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg 7081 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac 7141 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg 7201 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt 7261 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg 7321 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc 7381 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt 7441 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt 7501 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag 7561 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt 7621 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc 7681 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc 7741 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg 7801 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac 7861 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg 7921 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc 7981 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact 8041 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc 8101 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat 8161 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc 8221 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac 8281 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa 8341 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact 8401 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg 8461 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg 8521 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag 8581 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca 8641 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc 8701 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc 8761 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag 8821 gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg 8881 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg 8941 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtgc 9001 caagctg //