LOCUS V034643 9067 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V034643 VERSION V034643 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..3093 /label="PCGF1(NM_032673)" /note="PCGF1(NM_032673)" /gene="PCGF1" CDS 3100..3165 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3209..3420 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3433..3701 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 3730..3739 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 3760..4422 /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 4492..4545 /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 4546..5145 /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 5146..5734 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5808..6041 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6113..6234 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6274..6409 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6442..6458) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6466..6482 /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(6490..6520) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6535..6556 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6844..7432) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7603..8463) /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(8464..8568) /gene="bla" /label="AmpR promoter" primer_bind 9042..9058 /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 cgtctcctca ggggggccag 2341 attgcgatcg cgatgaggct tcggaaccag ctccagtcag tgtacaagat ggacccgcta 2401 cggaacgagg aggaggttcg agtgaagatc aaagacttga atgaacacat tgtttgctgc 2461 ctatgcgccg gctacttcgt ggatgccacc accatcacag agtgtcttca tactttctgc 2521 aagagttgta ttgtgaagta cctccaaact agcaagtact gccccatgtg caacattaag 2581 atccacgaga cacagccact gctcaacctc aaactggacc gggtcatgca ggacatcgtg 2641 tataagctgg tgcctggctt gcaagacagt gaagagaaac ggattcggga attctaccag 2701 tcccgaggtt tggaccgggt cacccagccc actggggaag agccagcact gagcaacctc 2761 ggcctcccct tcagcagctt tgaccactct aaagcccact actatcgcta tgatgagcag 2821 ttgaacctgt gcctggagcg gctgagttct ggcaaagaca agaataaaag cgtcctgcag 2881 aacaagtatg tccgatgttc tgttagagct gaggtacgcc atctccggag ggtcctgtgt 2941 caccgcttga tgctaaaccc tcagcatgtg cagctccttt ttgacaatga agttctccct 3001 gatcacatga caatgaagca gatatggctc tcccgctggt tcggcaagcc atcccctttg 3061 cttttacaat acagtgtgaa agagaagagg aggggatccg actacaaaga ccatgacggt 3121 gattataaag atcatgacat cgactacaag gatgacgatg acaagtagcg gccgcgaagg 3181 atctgcgatc gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag 3241 aagttggggg gaggggtcgg caattgaacg ggtgcctaga gaaggtggcg cggggtaaac 3301 tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat 3361 ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag 3421 ctgaagcttc gaggggctcg catctctcct tcacgcgccc gccgccctac ctgaggccgc 3481 catccacgcc ggttgagtcg cgttctgccg cctcccgcct gtggtgcctc ctgaactgcg 3541 tccgccgtct aggtaagttt aaagctcagg tcgagaccgg gcctttgtcc ggcgctccct 3601 tggagcctac ctagactcag ccggctctcc acgctttgcc tgaccctgct tgctcaactc 3661 tacgtctttg tttcgttttc tgttctgcgc cgttacagat ccaagctgtg accggcgcct 3721 acgctagacg ccaccatgga gagcgacgag agcggcctgc ccgccatgga gatcgagtgc 3781 cgcatcaccg gcaccctgaa cggcgtggag ttcgagctgg tgggcggcgg agagggcacc 3841 cccaagcagg gccgcatgac caacaagatg aagagcacca aaggcgccct gaccttcagc 3901 ccctacctgc tgagccacgt gatgggctac ggcttctacc acttcggcac ctaccccagc 3961 ggctacgaga accccttcct gcacgccatc aacaacggcg gctacaccaa cacccgcatc 4021 gagaagtacg aggacggcgg cgtgctgcac gtgagcttca gctaccgcta cgaggccggc 4081 cgcgtgatcg gcgacttcaa ggtggtgggc accggcttcc ccgaggacag cgtgatcttc 4141 accgacaaga tcatccgcag caacgccacc gtggagcacc tgcaccccat gggcgataac 4201 gtgctggtgg gcagcttcgc ccgcaccttc agcctgcgcg acggcggcta ctacagcttc 4261 gtggtggaca gccacatgca cttcaagagc gccatccacc ccagcatcct gcagaacggg 4321 ggccccatgt tcgccttccg ccgcgtggag gagctgcaca gcaacaccga gctgggcatc 4381 gtggagtacc agcacgcctt caagaccccc atcgccttcg ccagatcccg cgctcagtcg 4441 tccaattctg ccgtggacgg caccgccgga cccggctcca ccggatctcg cgagggcaga 4501 ggaagtcttc taacatgcgg tgacgtggag gagaatcccg gccctatgac cgagtacaag 4561 cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc 4621 gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg 4681 gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg 4741 gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg 4801 gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg 4861 cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg 4921 gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc 4981 cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc 5041 cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc 5101 gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgaaatca acctctggat 5161 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt 5221 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc 5281 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg 5341 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc 5401 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa 5461 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat 5521 tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc 5581 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt 5641 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag 5701 acgagtcgga tctccctttg ggccgcctcc ccgcctggta cctttaagac caatgactta 5761 caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat 5821 tcactcccaa cgaagataag atctgctttt tgcttgtact gggtctctct ggttagacca 5881 gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag 5941 cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag 6001 atccctcaga cccttttagt cagtgtggaa aatctctagc agtagtagtt catgtcatct 6061 tattattcag tatttataac ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt 6121 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc 6181 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt 6241 ctggctctag ctatcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc 6301 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc 6361 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct agacttttgc 6421 agagaccaaa ttcgtaatca tgtcatagct gtttcctgtg tgaaattgtt atccgctcac 6481 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt 6541 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc 6601 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg 6661 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt 6721 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa 6781 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc 6841 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag 6901 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt 6961 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg 7021 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg 7081 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg 7141 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac 7201 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg 7261 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt 7321 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg 7381 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc 7441 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt 7501 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt 7561 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag 7621 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt 7681 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc 7741 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc 7801 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg 7861 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac 7921 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg 7981 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc 8041 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact 8101 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc 8161 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat 8221 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc 8281 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac 8341 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa 8401 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact 8461 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg 8521 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg 8581 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag 8641 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca 8701 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc 8761 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc 8821 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag 8881 gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg 8941 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg 9001 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtgc 9061 caagctg //