LOCUS V044500 9187 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V044500 VERSION V044500 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..3213 /label="ZDHHC3(NM_001135179)" /note="ZDHHC3(NM_001135179)" /gene="ZDHHC3" CDS 3220..3285 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3329..3540 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3553..3821 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 3850..3859 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 3880..4542 /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 4612..4665 /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 4666..5265 /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 5266..5854 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5928..6161 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6233..6354 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6394..6529 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6562..6578) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6586..6602 /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(6610..6640) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6655..6676 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6964..7552) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7723..8583) /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(8584..8688) /gene="bla" /label="AmpR promoter" primer_bind 9162..9178 /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 tgcttatccc cacccaccac 2341 ttccgaaaca ttgagcggaa accagaatac ctccagccag agaagtgtgt cccacccccc 2401 taccctggtc ctgtgggaac catgtggttt atccgtgacg gctgtggcat cgcctgtgcc 2461 atcgttacct ggtttctggt cctctatgcg gagttcgtgg tcctctttgt catgctgatt 2521 ccatctcgag actacgtgta tagcatcatc aacggaattg tgttcaacct gctggccttc 2581 ttggccctgg cctcccactg ccgggccatg ctgacggacc ccggggcagt gcccaaagga 2641 aatgccacta aagaattcat cgagagttta cagttgaagc ctgggcaggt ggtgtacaag 2701 tgccccaaat gctgcagcat caagcccgac cgagcccacc actgcagtgt ttgtaagcgg 2761 tgcattcgga agatggacca ccactgtccc tgggtcaaca actgtgtagg cgagaacaac 2821 cagaagtact tcgtcctgtt tacaatgtac atagctctca tttccttgca cgccctcatc 2881 atggtgggat tccacttcct gcattgcttt gaagaagatt ggacaaagtg cagctccttc 2941 tctccaccca ccacagtgat tctccttatc ctgctgtgct ttgagggcct gctcttcctc 3001 attttcacat cagtgatgtt tgggacccag gtgcactcca tctgcacaga tgagacggga 3061 atagaacaat tgaaaaagga agagagaaga tgggctaaaa aaacaaaatg gatgaacatg 3121 aaagccgttt ttggccaccc cttctctcta ggctgggcca gcccctttgc cacgccagac 3181 caagggaagg cagacccgta ccagtatgtg gtcggatccg actacaaaga ccatgacggt 3241 gattataaag atcatgacat cgactacaag gatgacgatg acaagtagcg gccgcgaagg 3301 atctgcgatc gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag 3361 aagttggggg gaggggtcgg caattgaacg ggtgcctaga gaaggtggcg cggggtaaac 3421 tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat 3481 ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag 3541 ctgaagcttc gaggggctcg catctctcct tcacgcgccc gccgccctac ctgaggccgc 3601 catccacgcc ggttgagtcg cgttctgccg cctcccgcct gtggtgcctc ctgaactgcg 3661 tccgccgtct aggtaagttt aaagctcagg tcgagaccgg gcctttgtcc ggcgctccct 3721 tggagcctac ctagactcag ccggctctcc acgctttgcc tgaccctgct tgctcaactc 3781 tacgtctttg tttcgttttc tgttctgcgc cgttacagat ccaagctgtg accggcgcct 3841 acgctagacg ccaccatgga gagcgacgag agcggcctgc ccgccatgga gatcgagtgc 3901 cgcatcaccg gcaccctgaa cggcgtggag ttcgagctgg tgggcggcgg agagggcacc 3961 cccaagcagg gccgcatgac caacaagatg aagagcacca aaggcgccct gaccttcagc 4021 ccctacctgc tgagccacgt gatgggctac ggcttctacc acttcggcac ctaccccagc 4081 ggctacgaga accccttcct gcacgccatc aacaacggcg gctacaccaa cacccgcatc 4141 gagaagtacg aggacggcgg cgtgctgcac gtgagcttca gctaccgcta cgaggccggc 4201 cgcgtgatcg gcgacttcaa ggtggtgggc accggcttcc ccgaggacag cgtgatcttc 4261 accgacaaga tcatccgcag caacgccacc gtggagcacc tgcaccccat gggcgataac 4321 gtgctggtgg gcagcttcgc ccgcaccttc agcctgcgcg acggcggcta ctacagcttc 4381 gtggtggaca gccacatgca cttcaagagc gccatccacc ccagcatcct gcagaacggg 4441 ggccccatgt tcgccttccg ccgcgtggag gagctgcaca gcaacaccga gctgggcatc 4501 gtggagtacc agcacgcctt caagaccccc atcgccttcg ccagatcccg cgctcagtcg 4561 tccaattctg ccgtggacgg caccgccgga cccggctcca ccggatctcg cgagggcaga 4621 ggaagtcttc taacatgcgg tgacgtggag gagaatcccg gccctatgac cgagtacaag 4681 cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc 4741 gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg 4801 gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg 4861 gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg 4921 gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg 4981 cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg 5041 gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc 5101 cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc 5161 cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc 5221 gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgaaatca acctctggat 5281 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt 5341 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc 5401 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg 5461 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc 5521 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa 5581 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat 5641 tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc 5701 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt 5761 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag 5821 acgagtcgga tctccctttg ggccgcctcc ccgcctggta cctttaagac caatgactta 5881 caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat 5941 tcactcccaa cgaagataag atctgctttt tgcttgtact gggtctctct ggttagacca 6001 gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag 6061 cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag 6121 atccctcaga cccttttagt cagtgtggaa aatctctagc agtagtagtt catgtcatct 6181 tattattcag tatttataac ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt 6241 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc 6301 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt 6361 ctggctctag ctatcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc 6421 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc 6481 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct agacttttgc 6541 agagaccaaa ttcgtaatca tgtcatagct gtttcctgtg tgaaattgtt atccgctcac 6601 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt 6661 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc 6721 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg 6781 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt 6841 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa 6901 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc 6961 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag 7021 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt 7081 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg 7141 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg 7201 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg 7261 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac 7321 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg 7381 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt 7441 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg 7501 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc 7561 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt 7621 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt 7681 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag 7741 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt 7801 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc 7861 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc 7921 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg 7981 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac 8041 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg 8101 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc 8161 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact 8221 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc 8281 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat 8341 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc 8401 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac 8461 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa 8521 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact 8581 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg 8641 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg 8701 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag 8761 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca 8821 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc 8881 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc 8941 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag 9001 gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg 9061 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg 9121 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtgc 9181 caagctg //