LOCUS V040914 8804 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V040914 VERSION V040914 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..2805 /label="Skp1(NM_011543)" /note="Skp1(NM_011543)" /gene="Skp1" CDS 2812..2838 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" CDS 2842..2868 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" CDS 2872..2898 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" promoter 2963..3174 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3187..3455 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 3484..3493 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 3514..4176 /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 4246..4299 /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 4300..4899 /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 4900..5487 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5561..5794 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 5866..5987 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6006..6141 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6179..6195) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6203..6219 /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(6227..6257) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6272..6293 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6581..7169) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7340..8200) /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(8201..8305) /gene="bla" /label="AmpR promoter" primer_bind 8779..8795 /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 gccaccatgc ctacgataaa gttgcagagt 2341 tctgatggag agatatttga agttgatgta gaaattgcca aacaatctgt gactatcaag 2401 accatgctgg aagatttggg aatggatgat gaaggagatg atgatcctgt tcctttacca 2461 aatgttaatg cagcaattct aaagaaggtc attcagtggt gcacccacca caaagatgac 2521 cctcctcctc ctgaggatga tgaaaacaaa gaaaagcgga cagatgatat tcctgtttgg 2581 gaccaagaat tcctgaaagt tgaccaagga acactttttg aacttattct ggctgcaaac 2641 tacttagaca tcaaaggttt gcttgatgtc acatgcaaga ctgtggccaa tatgattaag 2701 gggaaaactc ctgaggagat tcgtaaaacc ttcaatatca aaaatgactt tactgaagag 2761 gaagaggccc aggtacgcaa agagaaccaa tggtgtgaag agaagctcga gtacccatac 2821 gacgtacctg attatgcaac ttatccttat gatgtacctg attatgctag ctatccttat 2881 gatgtcccag actacgcata gggatccctg cgatatccag cgcggccgcg aaggatctgc 2941 gatcgctccg gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc cgagaagttg 3001 gggggagggg tcggcaattg aacgggtgcc tagagaaggt ggcgcggggt aaactgggaa 3061 agtgatgtcg tgtactggct ccgccttttt cccgagggtg ggggagaacc gtatataagt 3121 gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg ccgccagaac acagctgaag 3181 cttcgagggg ctcgcatctc tccttcacgc gcccgccgcc ctacctgagg ccgccatcca 3241 cgccggttga gtcgcgttct gccgcctccc gcctgtggtg cctcctgaac tgcgtccgcc 3301 gtctaggtaa gtttaaagct caggtcgaga ccgggccttt gtccggcgct cccttggagc 3361 ctacctagac tcagccggct ctccacgctt tgcctgaccc tgcttgctca actctacgtc 3421 tttgtttcgt tttctgttct gcgccgttac agatccaagc tgtgaccggc gcctacgcta 3481 gacgccacca tggagagcga cgagagcggc ctgcccgcca tggagatcga gtgccgcatc 3541 accggcaccc tgaacggcgt ggagttcgag ctggtgggcg gcggagaggg cacccccaag 3601 cagggccgca tgaccaacaa gatgaagagc accaaaggcg ccctgacctt cagcccctac 3661 ctgctgagcc acgtgatggg ctacggcttc taccacttcg gcacctaccc cagcggctac 3721 gagaacccct tcctgcacgc catcaacaac ggcggctaca ccaacacccg catcgagaag 3781 tacgaggacg gcggcgtgct gcacgtgagc ttcagctacc gctacgaggc cggccgcgtg 3841 atcggcgact tcaaggtggt gggcaccggc ttccccgagg acagcgtgat cttcaccgac 3901 aagatcatcc gcagcaacgc caccgtggag cacctgcacc ccatgggcga taacgtgctg 3961 gtgggcagct tcgcccgcac cttcagcctg cgcgacggcg gctactacag cttcgtggtg 4021 gacagccaca tgcacttcaa gagcgccatc caccccagca tcctgcagaa cgggggcccc 4081 atgttcgcct tccgccgcgt ggaggagctg cacagcaaca ccgagctggg catcgtggag 4141 taccagcacg ccttcaagac ccccatcgcc ttcgccagat cccgcgctca gtcgtccaat 4201 tctgccgtgg acggcaccgc cggacccggc tccaccggat ctcgcgaggg cagaggaagt 4261 cttctaacat gcggtgacgt ggaggagaat cccggcccta tgaccgagta caagcccacg 4321 gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc 4381 gccgactacc ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc 4441 gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg 4501 gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg 4561 ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa 4621 cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc 4681 gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga 4741 gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac 4801 ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga 4861 ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgaa atcaacctct ggattacaaa 4921 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac 4981 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc 5041 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt 5101 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc 5161 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc 5221 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg 5281 gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt 5341 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc 5401 cgcggcctgc tgccggctct gcggcctctt ccgcgtctcc gccttcgccc tcagacgagt 5461 cggatctccc tttggccgcc tccccgcctg gtacctttaa gaccaatgac ttacaaggca 5521 gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc 5581 caacgaaaat aagatctgct ttttgcttgt actgggtctc tctggttaga ccagatctga 5641 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct 5701 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc 5761 agaccctttt agtcagtgtg gaaaatctct agcagtagta gttcatgtca tcttattatt 5821 cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggaactt gtttattgca 5881 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt 5941 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggctc 6001 tagctatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa 6061 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt 6121 gaggaggctt ttttggaggc ctagactttt gcagagacgg cccaaattcg taatcatggt 6181 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg 6241 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt 6301 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg 6361 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg 6421 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa 6481 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc 6541 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc 6601 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat 6661 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc 6721 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct 6781 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg 6841 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc 6901 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga 6961 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa 7021 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta 7081 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc 7141 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg 7201 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga 7261 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg 7321 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct 7381 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg 7441 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc 7501 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa 7561 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc 7621 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt 7681 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc 7741 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt 7801 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc 7861 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt 7921 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata 7981 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga 8041 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag 8101 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa 8161 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt 8221 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga 8281 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag 8341 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc 8401 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca 8461 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg 8521 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc 8581 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc 8641 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat 8701 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt 8761 tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa gctg //