LOCUS V047485 8551 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V047485 VERSION V047485 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..2577 /label="Ndufa4l2(NM_001271272)" /note="Ndufa4l2(NM_001271272)" /gene="Ndufa4l2" CDS 2584..2649 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 2693..2904 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 2917..3185 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 3214..3223 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 3244..3906 /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 3976..4029 /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 4030..4629 /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 4630..5218 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5292..5525 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 5597..5718 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 5758..5893 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(5926..5942) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5950..5966 /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(5974..6004) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6019..6040 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6328..6916) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7087..7947) /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(7948..8052) /gene="bla" /label="AmpR promoter" primer_bind 8526..8542 /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 caggaaccag tctaggggcc 2341 cgcttctacc ggcagataaa aagacaccct gggctcatcc caatgatcgg cttcatctgc 2401 ttgggcatgg gcagtgccgg actctacttg ctgcggcttg cccttcgcag ccctgatgtc 2461 tgctgggaca gaaagaacaa cccagagccc tggaaccgca tgagtcccaa tgaccagtac 2521 aagttccttg ctgtttccac cgactacaag aagctgaaga aggaccggcc tgacttcgga 2581 tccgactaca aagaccatga cggtgattat aaagatcatg acatcgacta caaggatgac 2641 gatgacaagt agcggccgcg aaggatctgc gatcgctccg gtgcccgtca gtgggcagag 2701 cgcacatcgc ccacagtccc cgagaagttg gggggagggg tcggcaattg aacgggtgcc 2761 tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg tgtactggct ccgccttttt 2821 cccgagggtg ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc 2881 aacgggtttg ccgccagaac acagctgaag cttcgagggg ctcgcatctc tccttcacgc 2941 gcccgccgcc ctacctgagg ccgccatcca cgccggttga gtcgcgttct gccgcctccc 3001 gcctgtggtg cctcctgaac tgcgtccgcc gtctaggtaa gtttaaagct caggtcgaga 3061 ccgggccttt gtccggcgct cccttggagc ctacctagac tcagccggct ctccacgctt 3121 tgcctgaccc tgcttgctca actctacgtc tttgtttcgt tttctgttct gcgccgttac 3181 agatccaagc tgtgaccggc gcctacgcta gacgccacca tggagagcga cgagagcggc 3241 ctgcccgcca tggagatcga gtgccgcatc accggcaccc tgaacggcgt ggagttcgag 3301 ctggtgggcg gcggagaggg cacccccaag cagggccgca tgaccaacaa gatgaagagc 3361 accaaaggcg ccctgacctt cagcccctac ctgctgagcc acgtgatggg ctacggcttc 3421 taccacttcg gcacctaccc cagcggctac gagaacccct tcctgcacgc catcaacaac 3481 ggcggctaca ccaacacccg catcgagaag tacgaggacg gcggcgtgct gcacgtgagc 3541 ttcagctacc gctacgaggc cggccgcgtg atcggcgact tcaaggtggt gggcaccggc 3601 ttccccgagg acagcgtgat cttcaccgac aagatcatcc gcagcaacgc caccgtggag 3661 cacctgcacc ccatgggcga taacgtgctg gtgggcagct tcgcccgcac cttcagcctg 3721 cgcgacggcg gctactacag cttcgtggtg gacagccaca tgcacttcaa gagcgccatc 3781 caccccagca tcctgcagaa cgggggcccc atgttcgcct tccgccgcgt ggaggagctg 3841 cacagcaaca ccgagctggg catcgtggag taccagcacg ccttcaagac ccccatcgcc 3901 ttcgccagat cccgcgctca gtcgtccaat tctgccgtgg acggcaccgc cggacccggc 3961 tccaccggat ctcgcgaggg cagaggaagt cttctaacat gcggtgacgt ggaggagaat 4021 cccggcccta tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc 4081 agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc 4141 gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc 4201 gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc 4261 acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag 4321 ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg 4381 cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag 4441 ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc 4501 gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc 4561 gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc 4621 ggtgcctgaa atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac 4681 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt 4741 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat 4801 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca 4861 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc 4921 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg 4981 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct 5041 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct 5101 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt 5161 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct 5221 ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga 5281 aaagggggga ctggaagggc taattcactc ccaacgaaga taagatctgc tttttgcttg 5341 tactgggtct ctctggttag accagatctg agcctgggag ctctctggct aactagggaa 5401 cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct 5461 gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc 5521 tagcagtagt agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat 5581 atcagagagt gagaggaact tgtttattgc agcttataat ggttacaaat aaagcaatag 5641 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa 5701 actcatcaat gtatcttatc atgtctggct ctagctatcc cgcccctaac tccgcccatc 5761 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt 5821 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc 5881 ttttttggag gcctagactt ttgcagagac caaattcgta atcatgtcat agctgtttcc 5941 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg 6001 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc 6061 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg 6121 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc 6181 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac 6241 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa 6301 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca 6361 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc 6421 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata 6481 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta 6541 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca 6601 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga 6661 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg 6721 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg 6781 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg 6841 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag 6901 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa 6961 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat 7021 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc 7081 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc 7141 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc 7201 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc 7261 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc 7321 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt 7381 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc 7441 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa 7501 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt 7561 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg 7621 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc 7681 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa 7741 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt 7801 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt 7861 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag 7921 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta 7981 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat 8041 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat 8101 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg 8161 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta 8221 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg 8281 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg 8341 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag 8401 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc 8461 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg 8521 acgttgtaaa acgacggcca gtgccaagct g //