LOCUS V045297 9511 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V045297 VERSION V045297 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..1920 /note="central polypurine tract and central termination sequence of HIV-1" /label="cPPT/CTS" promoter 2010..2213 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 2302..3018 /codon_start=1 /note="mammalian codon-optimized" /product="the original enhanced GFP (Yang et al., 1996)" /transl_table=1 /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL TLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV LLEFVTAAGITLGMDELYK" /label="EGFP" CDS 3055..4365 /label="PLIN2(NM_001122)" /note="PLIN2(NM_001122)" /gene="PLIN2" CDS 4372..4437 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 4481..4692 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4705..4973 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 5000..5599 /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 5606..6194 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6268..6501 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6573..6694 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6713..6848 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6886..6902) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6910..6926 /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(6934..6964) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6979..7000 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7288..7876) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8047..8907) /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(8908..9012) /gene="bla" /label="AmpR promoter" primer_bind 9486..9502 /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 attacaaaat tcaaaatttt 1921 atcgatacta gtattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca 1981 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc 2041 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga 2101 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat 2161 tgacgcaaat gggcggtagg cgtgtacggt gggaggttta tataagcaga gctcgtttag 2221 tgaccgtcag atcgcctgga gacgccatcc acgctgtttt gacctccata gaagattcta 2281 gagctagcga attcagccac catggtgagc aagggcgagg agctgttcac cggggtggtg 2341 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag 2401 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag 2461 ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc 2521 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac 2581 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg 2641 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag 2701 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc 2761 atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag 2821 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc 2881 gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac 2941 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc 3001 atggacgagc tgtacaaggg aggaggggga agcggtggag gcggttctct cgagatggca 3061 tccgttgcag ttgatccaca accgagtgtg gtgactcggg tggtcaacct gcccttggtg 3121 agctccacgt atgacctcat gtcctcagcc tatctcagta caaaggacca gtatccctac 3181 ctgaagtctg tgtgtgagat ggcagagaac ggtgtgaaga ccatcacctc cgtggccatg 3241 accagtgctc tgcccatcat ccagaagcta gagccgcaaa ttgcagttgc caatacctat 3301 gcctgtaagg ggctagacag gattgaggag agactgccta ttctgaatca gccatcaact 3361 cagattgttg ccaatgccaa aggcgctgtg actggggcaa aagatgctgt gacgactact 3421 gtgactgggg ccaaggattc tgtggccagc acgatcacag gggtgatgga caagaccaaa 3481 ggggcagtga ctggcagtgt ggagaagacc aagtctgtgg tcagtggcag cattaacaca 3541 gtcttgggga gtcggatgat gcagctcgtg agcagtggcg tagaaaatgc actcaccaaa 3601 tcagagctgt tggtagaaca gtacctccct ctcactgagg aagaactaga aaaagaagca 3661 aaaaaagttg aaggatttga tctggttcag aagccaagtt attatgttag actgggatcc 3721 ctgtctacca agcttcactc ccgtgcctac cagcaggctc tcagcagggt taaagaagct 3781 aagcaaaaaa gccaacagac catttctcag ctccattcta ctgttcacct gattgaattt 3841 gccaggaaga atgtgtatag tgccaatcag aaaattcagg atgctcagga taagctctac 3901 ctctcatggg tagagtggaa aaggagcatt ggatatgatg atactgatga gtcccactgt 3961 gctgagcaca ttgagtcacg tactcttgca attgcccgca acctgactca gcagctccag 4021 accacgtgcc acaccctcct gtccaacatc caaggtgtac cacagaacat ccaagatcaa 4081 gccaagcaca tgggggtgat ggcaggcgac atctactcag tgttccgcaa tgctgcctcc 4141 tttaaagaag tgtctgacag cctcctcact tctagcaagg ggcagctgca gaaaatgaag 4201 gaatctttag atgacgtgat ggattatctt gttaacaaca cgcccctcaa ctggctggta 4261 ggtccctttt atcctcagct gactgagtct cagaatgctc aggaccaagg tgcagagatg 4321 gacaagagca gccaggagac ccagcgatct gagcataaaa ctcatggatc cgactacaaa 4381 gaccatgacg gtgattataa agatcatgac atcgactaca aggatgacga tgacaagtag 4441 cggccgcgaa ggatctgcga tcgctccggt gcccgtcagt gggcagagcg cacatcgccc 4501 acagtccccg agaagttggg gggaggggtc ggcaattgaa cgggtgccta gagaaggtgg 4561 cgcggggtaa actgggaaag tgatgtcgtg tactggctcc gcctttttcc cgagggtggg 4621 ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc 4681 gccagaacac agctgaagct tcgaggggct cgcatctctc cttcacgcgc ccgccgccct 4741 acctgaggcc gccatccacg ccggttgagt cgcgttctgc cgcctcccgc ctgtggtgcc 4801 tcctgaactg cgtccgccgt ctaggtaagt ttaaagctca ggtcgagacc gggcctttgt 4861 ccggcgctcc cttggagcct acctagactc agccggctct ccacgctttg cctgaccctg 4921 cttgctcaac tctacgtctt tgtttcgttt tctgttctgc gccgttacag atccaagctg 4981 tgaccggcgc ctacgctaga tgaccgagta caagcccacg gtgcgcctcg ccacccgcga 5041 cgacgtcccc cgggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg 5101 ccacaccgtc gacccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct 5161 cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc 5221 ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg 5281 catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc 5341 gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca 5401 ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc 5461 cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct 5521 cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac 5581 ccgcaagccc ggtgcctgag tcgacaatca acctctggat tacaaaattt gtgaaagatt 5641 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc 5701 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg 5761 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac 5821 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc 5881 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc 5941 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa 6001 atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc 6061 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc 6121 ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg 6181 ggccgcctcc ccgcctggta cctttaagac caatgactta caaggcagct gtagatctta 6241 gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa cgaaaataag 6301 atctgctttt tgcttgtact gggtctctct ggttagacca gatctgagcc tgggagctct 6361 ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag 6421 tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt 6481 cagtgtggaa aatctctagc agtagtagtt catgtcatct tattattcag tatttataac 6541 ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt tattgcagct tataatggtt 6601 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta 6661 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggctctag ctatcccgcc 6721 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta 6781 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt 6841 tggaggccta gacttttgca gagacggccc aaattcgtaa tcatggtcat agctgtttcc 6901 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg 6961 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc 7021 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg 7081 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc 7141 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac 7201 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa 7261 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca 7321 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc 7381 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata 7441 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta 7501 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca 7561 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga 7621 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg 7681 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg 7741 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg 7801 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag 7861 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa 7921 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat 7981 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc 8041 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc 8101 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc 8161 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc 8221 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc 8281 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt 8341 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc 8401 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa 8461 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt 8521 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg 8581 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc 8641 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa 8701 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt 8761 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt 8821 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag 8881 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta 8941 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat 9001 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat 9061 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg 9121 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta 9181 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg 9241 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg 9301 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag 9361 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc 9421 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg 9481 acgttgtaaa acgacggcca gtgccaagct g //