LOCUS V043683 9487 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V043683 VERSION V043683 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..4341 /label="ZBP1(NM_030776)" /note="ZBP1(NM_030776)" /gene="ZBP1" CDS 4348..4413 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 4457..4668 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4681..4949 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 4976..5575 /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 5582..6170 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6244..6477 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6549..6670 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6689..6824 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6862..6878) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6886..6902 /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(6910..6940) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6955..6976 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7264..7852) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8023..8883) /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(8884..8988) /gene="bla" /label="AmpR promoter" primer_bind 9462..9478 /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 cgagatggcc 3061 caggctcctg ctgacccggg cagagaaggc caccttgaac aaagaatcct gcaggtgctg 3121 acagaggctg gctccccggt gaaacttgcc cagctggtga aggaatgcca agcacccaag 3181 agggagctca accaagtcct ctaccgaatg aaaaaggagt tgaaagtctc cctcacatcc 3241 cctgccacct ggtgcttggg cgggactgat cctgaaggcg agggtcctgc agagctggcc 3301 ttgtccagcc ctgccgagag gccccagcaa catgcagcta caattccaga gacccctggc 3361 cctcagttca gccaacaacg ggaggaagac atctacaggt ttctcaaaga caatggtccc 3421 cagagggccc tggtcatcgc ccaagcactg ggaatgagga cagcaaaaga tgtgaaccga 3481 gacttgtaca ggatgaagag caggcacctt ctggacatgg atgagcagtc caaagcatgg 3541 acgatttacc gcccagaaga ttctggaaga agagcaaagt cagcctcaat tatttaccag 3601 cacaatccaa tcaacatgat ctgccagaat ggacccaaca gctggatttc cattgcaaac 3661 tccgaagcca tccagattgg acacgggaac atcattacaa gacagacagt ctccagggag 3721 gacggttccg ccggtccacg ccacctccct tcaatggcac caggtgattc ctcaacttgg 3781 gggaccctag ttgatccctg ggggccccag gacatccaca tggagcagtc catactgaga 3841 cgggtgcagc tgggacacag caatgagatg aggctccacg gcgtcccgtc cgagggccct 3901 gcccacatcc cccctggcag ccccccagtc tctgccactg ctgccggccc agaagcttcg 3961 tttgaagcaa gaattcccag tccaggaact caccctgagg gggaagccgc ccagagaatc 4021 cacatgaaat cgtgctttct cgaggacgcc accatcggca acagcaacaa aatgtctatc 4081 agcccagggg tggctggccc aggaggagtc gcagggtctg gagaggggga gccaggggag 4141 gacgcaggtc gtcgtcccgc agacacacaa tccagaagtc actttcctcg agacattggt 4201 cagcccatca ctcccagcca ctcgaagctc acccccaagc tggaaactat gactcttgga 4261 aacaggagtc acaaagctgc agaaggcagc cactatgtgg atgaagcctc acacgagggg 4321 agctggtggg gaggtgggat tggatccgac tacaaagacc atgacggtga ttataaagat 4381 catgacatcg actacaagga tgacgatgac aagtagcggc cgcgaaggat ctgcgatcgc 4441 tccggtgccc gtcagtgggc agagcgcaca tcgcccacag tccccgagaa gttgggggga 4501 ggggtcggca attgaacggg tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat 4561 gtcgtgtact ggctccgcct ttttcccgag ggtgggggag aaccgtatat aagtgcagta 4621 gtcgccgtga acgttctttt tcgcaacggg tttgccgcca gaacacagct gaagcttcga 4681 ggggctcgca tctctccttc acgcgcccgc cgccctacct gaggccgcca tccacgccgg 4741 ttgagtcgcg ttctgccgcc tcccgcctgt ggtgcctcct gaactgcgtc cgccgtctag 4801 gtaagtttaa agctcaggtc gagaccgggc ctttgtccgg cgctcccttg gagcctacct 4861 agactcagcc ggctctccac gctttgcctg accctgcttg ctcaactcta cgtctttgtt 4921 tcgttttctg ttctgcgccg ttacagatcc aagctgtgac cggcgcctac gctagatgac 4981 cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtcccccggg ccgtacgcac 5041 cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgacc cggaccgcca 5101 catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg 5161 caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt 5221 cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg 5281 gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc 5341 gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc 5401 cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac 5461 ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt 5521 cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgagtcga 5581 caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc 5641 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg 5701 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt 5761 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac 5821 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc 5881 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct 5941 gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct 6001 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct 6061 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct 6121 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggtacctt 6181 taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg 6241 gactggaagg gctaattcac tcccaacgaa aataagatct gctttttgct tgtactgggt 6301 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc 6361 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg 6421 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta 6481 gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga 6541 gtgagaggaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa 6601 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca 6661 atgtatctta tcatgtctgg ctctagctat cccgccccta actccgccca gttccgccca 6721 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc 6781 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctagact tttgcagaga 6841 cggcccaaat tcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac 6901 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt 6961 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc 7021 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg 7081 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt 7141 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa 7201 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc 7261 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag 7321 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt 7381 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg 7441 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg 7501 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg 7561 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac 7621 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg 7681 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt 7741 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg 7801 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc 7861 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt 7921 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt 7981 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag 8041 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt 8101 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc 8161 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc 8221 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg 8281 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac 8341 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg 8401 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc 8461 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact 8521 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc 8581 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat 8641 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc 8701 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac 8761 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa 8821 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact 8881 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg 8941 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg 9001 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag 9061 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca 9121 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc 9181 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc 9241 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag 9301 gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg 9361 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg 9421 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtgc 9481 caagctg //