LOCUS V045161 8611 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V045161 VERSION V045161 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..3465 /label="DGAT2(NM_032564)" /note="DGAT2(NM_032564)" /gene="DGAT2" CDS 3472..3537 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3581..3792 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3805..4073 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 4100..4699 /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 4706..5294 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5368..5601 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 5673..5794 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 5813..5948 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(5986..6002) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6010..6026 /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(6034..6064) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6079..6100 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6388..6976) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7147..8007) /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(8008..8112) /gene="bla" /label="AmpR promoter" primer_bind 8586..8602 /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 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagattct 2281 agagctagcg aattcgccac catgaagacc ctcatagccg cctactccgg ggtcctgcgc 2341 ggcgagcgtc aggccgaggc tgaccggagc cagcgctctc acggaggacc tgcgctgtcg 2401 cgcgaggggt ctgggagatg gggcactgga tccagcatcc tctccgccct ccaggacctc 2461 ttctctgtca cctggctcaa taggtccaag gtggaaaagc agctacaggt catctcagtg 2521 ctccagtggg tcctgtcctt ccttgtactg ggagtggcct gcagtgccat cctcatgtac 2581 atattctgca ctgattgctg gctcatcgct gtgctctact tcacttggct ggtgtttgac 2641 tggaacacac ccaagaaagg tggcaggagg tcacagtggg tccgaaactg ggctgtgtgg 2701 cgctactttc gagactactt tcccatccag ctggtgaaga cacacaacct gctgaccacc 2761 aggaactata tctttggata ccacccccat ggtatcatgg gcctgggtgc cttctgcaac 2821 ttcagcacag aggccacaga agtgagcaag aagttcccag gcatacggcc ttacctggct 2881 acactggcag gcaacttccg aatgcctgtg ttgagggagt acctgatgtc tggaggtatc 2941 tgccctgtca gccgggacac catagactat ttgctttcaa agaatgggag tggcaatgct 3001 atcatcatcg tggtcggggg tgcggctgag tctctgagct ccatgcctgg caagaatgca 3061 gtcaccctgc ggaaccgcaa gggctttgtg aaactggccc tgcgtcatgg agctgacctg 3121 gttcccatct actcctttgg agagaatgaa gtgtacaagc aggtgatctt cgaggagggc 3181 tcctggggcc gatgggtcca gaagaagttc cagaaataca ttggtttcgc cccatgcatc 3241 ttccatggtc gaggcctctt ctcctccgac acctgggggc tggtgcccta ctccaagccc 3301 atcaccactg ttgtgggaga gcccatcacc atccccaagc tggagcaccc aacccagcaa 3361 gacatcgacc tgtaccacac catgtacatg gaggccctgg tgaagctctt cgacaagcac 3421 aagaccaagt tcggcctccc ggagactgag gtcctggagg tgaacggatc cgactacaaa 3481 gaccatgacg gtgattataa agatcatgac atcgactaca aggatgacga tgacaagtag 3541 cggccgcgaa ggatctgcga tcgctccggt gcccgtcagt gggcagagcg cacatcgccc 3601 acagtccccg agaagttggg gggaggggtc ggcaattgaa cgggtgccta gagaaggtgg 3661 cgcggggtaa actgggaaag tgatgtcgtg tactggctcc gcctttttcc cgagggtggg 3721 ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc 3781 gccagaacac agctgaagct tcgaggggct cgcatctctc cttcacgcgc ccgccgccct 3841 acctgaggcc gccatccacg ccggttgagt cgcgttctgc cgcctcccgc ctgtggtgcc 3901 tcctgaactg cgtccgccgt ctaggtaagt ttaaagctca ggtcgagacc gggcctttgt 3961 ccggcgctcc cttggagcct acctagactc agccggctct ccacgctttg cctgaccctg 4021 cttgctcaac tctacgtctt tgtttcgttt tctgttctgc gccgttacag atccaagctg 4081 tgaccggcgc ctacgctaga tgaccgagta caagcccacg gtgcgcctcg ccacccgcga 4141 cgacgtcccc cgggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg 4201 ccacaccgtc gacccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct 4261 cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc 4321 ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg 4381 catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc 4441 gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca 4501 ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc 4561 cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct 4621 cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac 4681 ccgcaagccc ggtgcctgag tcgacaatca acctctggat tacaaaattt gtgaaagatt 4741 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc 4801 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg 4861 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac 4921 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc 4981 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc 5041 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa 5101 atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc 5161 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc 5221 ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg 5281 ggccgcctcc ccgcctggta cctttaagac caatgactta caaggcagct gtagatctta 5341 gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa cgaaaataag 5401 atctgctttt tgcttgtact gggtctctct ggttagacca gatctgagcc tgggagctct 5461 ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag 5521 tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt 5581 cagtgtggaa aatctctagc agtagtagtt catgtcatct tattattcag tatttataac 5641 ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt tattgcagct tataatggtt 5701 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta 5761 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggctctag ctatcccgcc 5821 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta 5881 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt 5941 tggaggccta gacttttgca gagacggccc aaattcgtaa tcatggtcat agctgtttcc 6001 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg 6061 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc 6121 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg 6181 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc 6241 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac 6301 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa 6361 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca 6421 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc 6481 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata 6541 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta 6601 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca 6661 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga 6721 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg 6781 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg 6841 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg 6901 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag 6961 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa 7021 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat 7081 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc 7141 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc 7201 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc 7261 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc 7321 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc 7381 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt 7441 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc 7501 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa 7561 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt 7621 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg 7681 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc 7741 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa 7801 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt 7861 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt 7921 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag 7981 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta 8041 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat 8101 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat 8161 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg 8221 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta 8281 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg 8341 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg 8401 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag 8461 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc 8521 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg 8581 acgttgtaaa acgacggcca gtgccaagct g //