LOCUS V031553 9127 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V031553 VERSION V031553 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..3153 /label="ELOVL1(NM_022821)" /note="ELOVL1(NM_022821)" /gene="ELOVL1" CDS 3160..3225 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3269..3480 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3493..3761 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 3790..3799 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 3820..4482 /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 4552..4605 /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 4606..5205 /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 5206..5794 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5868..6101 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6173..6294 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6334..6469 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6502..6518) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6526..6542 /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(6550..6580) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6595..6616 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6904..7492) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7663..8523) /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(8524..8628) /gene="bla" /label="AmpR promoter" primer_bind 9102..9118 /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 aggctgttgt gaacttgtac 2341 caagaggtga tgaagcacgc agatccccgg atccagggct accctctgat ggggtccccc 2401 ttgctaatga cctccattct cctgacctac gtgtacttcg ttctctcact tgggcctcgc 2461 atcatggcta atcggaagcc cttccagctc cgtggcttca tgattgtcta caacttctca 2521 ctggtggcac tctccctcta cattgtctat gagttcctga tgtcgggctg gctgagcacc 2581 tatacctggc gctgtgaccc tgtggactat tccaacagcc ctgaggcact taggatggtt 2641 cgggtggcct ggctcttcct cttctccaag ttcattgagc tgatggacac agtgatcttt 2701 attctccgaa agaaagacgg gcaggtgacc ttcctacatg tcttccatca ctctgtgctt 2761 ccctggagct ggtggtgggg ggtaaagatt gccccgggag gaatgggctc tttccatgcc 2821 atgataaact cttccgtgca tgtcataatg tacctgtact acggattatc tgcctttggc 2881 cctgtggcac aaccctacct ttggtggaaa aagcacatga cagccattca gctgatccag 2941 tttgtcctgg tctcactgca catctcccag tactacttta tgtccagctg taactaccag 3001 tacccagtca ttattcacct catctggatg tatggcacca tcttcttcat gctgttctcc 3061 aacttctggt atcactctta taccaagggc aagcggctgc cccgtgcact tcagcaaaat 3121 ggagctccag gtattgccaa ggtcaaggcc aacggatccg actacaaaga ccatgacggt 3181 gattataaag atcatgacat cgactacaag gatgacgatg acaagtagcg gccgcgaagg 3241 atctgcgatc gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag 3301 aagttggggg gaggggtcgg caattgaacg ggtgcctaga gaaggtggcg cggggtaaac 3361 tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat 3421 ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag 3481 ctgaagcttc gaggggctcg catctctcct tcacgcgccc gccgccctac ctgaggccgc 3541 catccacgcc ggttgagtcg cgttctgccg cctcccgcct gtggtgcctc ctgaactgcg 3601 tccgccgtct aggtaagttt aaagctcagg tcgagaccgg gcctttgtcc ggcgctccct 3661 tggagcctac ctagactcag ccggctctcc acgctttgcc tgaccctgct tgctcaactc 3721 tacgtctttg tttcgttttc tgttctgcgc cgttacagat ccaagctgtg accggcgcct 3781 acgctagacg ccaccatgga gagcgacgag agcggcctgc ccgccatgga gatcgagtgc 3841 cgcatcaccg gcaccctgaa cggcgtggag ttcgagctgg tgggcggcgg agagggcacc 3901 cccaagcagg gccgcatgac caacaagatg aagagcacca aaggcgccct gaccttcagc 3961 ccctacctgc tgagccacgt gatgggctac ggcttctacc acttcggcac ctaccccagc 4021 ggctacgaga accccttcct gcacgccatc aacaacggcg gctacaccaa cacccgcatc 4081 gagaagtacg aggacggcgg cgtgctgcac gtgagcttca gctaccgcta cgaggccggc 4141 cgcgtgatcg gcgacttcaa ggtggtgggc accggcttcc ccgaggacag cgtgatcttc 4201 accgacaaga tcatccgcag caacgccacc gtggagcacc tgcaccccat gggcgataac 4261 gtgctggtgg gcagcttcgc ccgcaccttc agcctgcgcg acggcggcta ctacagcttc 4321 gtggtggaca gccacatgca cttcaagagc gccatccacc ccagcatcct gcagaacggg 4381 ggccccatgt tcgccttccg ccgcgtggag gagctgcaca gcaacaccga gctgggcatc 4441 gtggagtacc agcacgcctt caagaccccc atcgccttcg ccagatcccg cgctcagtcg 4501 tccaattctg ccgtggacgg caccgccgga cccggctcca ccggatctcg cgagggcaga 4561 ggaagtcttc taacatgcgg tgacgtggag gagaatcccg gccctatgac cgagtacaag 4621 cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc 4681 gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg 4741 gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg 4801 gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg 4861 gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg 4921 cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg 4981 gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc 5041 cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc 5101 cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc 5161 gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgaaatca acctctggat 5221 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt 5281 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc 5341 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg 5401 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc 5461 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa 5521 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat 5581 tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc 5641 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt 5701 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag 5761 acgagtcgga tctccctttg ggccgcctcc ccgcctggta cctttaagac caatgactta 5821 caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat 5881 tcactcccaa cgaagataag atctgctttt tgcttgtact gggtctctct ggttagacca 5941 gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag 6001 cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag 6061 atccctcaga cccttttagt cagtgtggaa aatctctagc agtagtagtt catgtcatct 6121 tattattcag tatttataac ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt 6181 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc 6241 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt 6301 ctggctctag ctatcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc 6361 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc 6421 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct agacttttgc 6481 agagaccaaa ttcgtaatca tgtcatagct gtttcctgtg tgaaattgtt atccgctcac 6541 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt 6601 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc 6661 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg 6721 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt 6781 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa 6841 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc 6901 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag 6961 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt 7021 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg 7081 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg 7141 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg 7201 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac 7261 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg 7321 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt 7381 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg 7441 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc 7501 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt 7561 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt 7621 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag 7681 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt 7741 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc 7801 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc 7861 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg 7921 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac 7981 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg 8041 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc 8101 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact 8161 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc 8221 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat 8281 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc 8341 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac 8401 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa 8461 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact 8521 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg 8581 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg 8641 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag 8701 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca 8761 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc 8821 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc 8881 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag 8941 gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg 9001 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg 9061 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtgc 9121 caagctg //