LOCUS V018463 9727 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V018463 VERSION V018463 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers LTR 1..634 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" misc_feature 681..806 /note="packaging signal of human immunodeficiency virus type 1" /label="HIV-1 Ψ" misc_feature 1303..1536 /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." /label="RRE" CDS 1721..1765 /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 2028..2143 /note="central polypurine tract and central termination sequence of HIV-1 (lacking the first T)" /label="cPPT/CTS" enhancer 2201..2504 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 2505..2708 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" regulatory 2809..2818 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 2818..2847 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /transl_table=1 /translation="EQKLISEEDL" /label="Myc" CDS 2854..4404 /label="TRIM65(NM_173547)" /note="TRIM65(NM_173547)" /gene="TRIM65" misc_feature 4430..5016 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" /label="IRES2" CDS 5036..5635 /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 5649..6237 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6444..7077 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(7205..7221) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7229..7245 /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(7253..7283) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7298..7319 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7607..8195) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8366..9226) /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(9227..9331) /gene="bla" /label="AmpR promoter" polyA_signal 9379..9513 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" ORIGIN 1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca 61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac 121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca 181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg 241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag 301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg 361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat 421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga 481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct 541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc 601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag 661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg 721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga 781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg 841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg 901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct 961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat 1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca 1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc 1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag 1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc 1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg 1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc 1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc 1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct 1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa 1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca 1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt 1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt 1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta 1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt 1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct 1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga 1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccttt aaaagaaaag 2041 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac 2101 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg 2161 acagcagaga tccagtttat cgataagctt gggagttccg cgttacataa cttacggtaa 2221 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg 2281 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt 2341 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg 2401 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc 2461 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc 2521 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca 2581 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta 2641 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa 2701 gcagagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc 2761 tccatagaag acaccgactc tactagagga tctatttccg gtgaattcgc caccatggaa 2821 caaaaactca tctcagaaga ggatctgctc gagatggccg cgcagctgct ggaggagaag 2881 ctgacctgcg ccatctgcct ggggctctac caggacccag tgacgctgcc ctgcggccac 2941 aacttctgcg gggcctgcat ccgggactgg tgggaccgct gcggaaaggc gtgccccgag 3001 tgccgggagc cctttcccga cggcgccgag ctgcgccgca acgtggccct cagcggcgtg 3061 ctggaggtgg tgcgcgccgg gcccgcccgg gatcccggcc ccgatcccgg ccccggcccc 3121 gaccctgccg cgcgctgccc ccgccacggg cggccgctgg agctcttctg ccggaccgag 3181 ggccgctgtg tgtgcagcgt gtgcaccgtg cgcgagtgtc gcctccacga gcgggcgctg 3241 ctggatgccg agcgcctcaa gcgcgaggcc cagctgagag ccagcctgga ggttacccag 3301 cagcaggcca cccaggccga aggccagcta ctagagctgc gcaagcaaag cagccagatc 3361 cagaactcgg cctgcatctt ggcctcctgg gtctccggca agttcagcag cctgctacag 3421 gccctggaaa tacagcacac gacagcactg aggagcatcg aggtggccaa gacgcaggcg 3481 ctggcacagg ctcgagacga ggagcagcgg ctgcgggtcc atttggaggc tgtggctcgc 3541 catggctgca ggatccggga gctcctggag caggtggatg agcagacctt cctgcaggaa 3601 tcgcagctcc tccagccccc agggcctctt gggccactga cccctctgca gtgggatgaa 3661 gaccaacagc tgggtgacct gaagcagttg ctaagccggc tgtgtggcct cctcttggaa 3721 gaggggagcc accctggggc accagccaag cctgtggact tagcccccgt ggaggcccca 3781 ggtcccctgg caccggtccc aagcacagtt tgtccactga ggaggaaact ctggcagaat 3841 tatcgcaatc tgacctttga tccagtcagc gccaaccgtc acttctatct gtcgcgccag 3901 gaccagcagg tgaagcactg tcgtcagtcc cggggcccag gcgggcccgg cagctttgag 3961 ctctggcagg tgcaatgtgc ccagagcttc caggccgggc accactactg ggaggtgcgc 4021 gcgtcagacc actcggtgac actgggcgtc tcctacccgc aactgccacg gtgcaggctg 4081 gggccccaca cagacaacat tggccgggga ccctgctcct gggggctctg cgtccaggag 4141 gacagcctcc aggcctggca caacggggaa gcccagcgcc tcccaggggt gtcagggcgg 4201 ctcctgggca tggatttgga cctggcctca ggctgcctca ccttctacag cctggagccc 4261 cagacccagc ccctgtacac cttccatgcc ctcttcaacc agcccctcac ccccgtcttc 4321 tggctcctcg agggtaggac cctgaccctg tgccatcagc caggggctgt gttccctctg 4381 gggccccagg aagaggtgct cagctaatct agagcggccg cggatcccgc ccctctccct 4441 cccccccccc taacgttact ggccgaagcc gcttggaata aggccggtgt gcgtttgtct 4501 atatgttatt ttccaccata ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc 4561 ctgtcttctt gacgagcatt cctaggggtc tttcccctct cgccaaagga atgcaaggtc 4621 tgttgaatgt cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa acaacgtctg 4681 tagcgaccct ttgcaggcag cggaaccccc cacctggcga caggtgcctc tgcggccaaa 4741 agccacgtgt ataagataca cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt 4801 ggatagttgt ggaaagagtc aaatggctct cctcaagcgt attcaacaag gggctgaagg 4861 atgcccagaa ggtaccccat tgtatgggat ctgatctggg gcctcggtgc acatgcttta 4921 catgtgttta gtcgaggtta aaaaaacgtc taggcccccc gaaccacggg gacgtggttt 4981 tcctttgaaa aacacgatga taagcttgcc acaacccaca aggagacgac cttccatgac 5041 cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtcccccggg ccgtacgcac 5101 cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgacc cggaccgcca 5161 catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg 5221 caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt 5281 cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg 5341 gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc 5401 gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc 5461 cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac 5521 ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt 5581 cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctagacgcg 5641 tctggaacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact 5701 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg 5761 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg 5821 aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa 5881 cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc 5941 ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg 6001 ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaagctgacg tcctttccat 6061 ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt 6121 cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc 6181 cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg 6241 gaattaattc tgcagtcgag acctagaaaa acatggagca atcacaagta gcaatacagc 6301 agctaccaat gctgattgtg cctggctaga agcacaagag gaggaggagg tgggttttcc 6361 agtcacacct caggtacctt taagaccaat gacttacaag gcagctgtag atcttagcca 6421 ctttttaaaa gaaaagaggg gactggaagg gctaattcac tcccaacgaa gacaagatat 6481 ccttgatctg tggatctacc acacacaagg ctacttccct gattagcaga actacacacc 6541 agggccaggg gtcagatatc cactgacctt tggatggtgc tacaagctag taccagttga 6601 gccagataag gtagaagagg ccaataaagg agagaacacc agcttgttac accctgtgag 6661 cctgcatggg atggatgacc cggagagaga agtgttagag tggaggtttg acagccgcct 6721 agcatttcat cacgtggccc gagagctgca tccggagtac ttcaagaact gctgatatcg 6781 agcttgctac aagggacttt ccgctgggga ctttccaggg aggcgtggcc tgggcgggac 6841 tggggagtgg cgagccctca gatcctgcat ataagcagct gctttttgcc tgtactgggt 6901 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc 6961 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg 7021 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta 7081 gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga 7141 gtgagaggcc ttgacattgc tagcgtttac cgtcgacctc tagctagagc ttggcgtaat 7201 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac 7261 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa 7321 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat 7381 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc 7441 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg 7501 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag 7561 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc 7621 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag 7681 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga 7741 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc 7801 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg 7861 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt 7921 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca 7981 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca 8041 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag 8101 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca 8161 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg 8221 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa 8281 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta 8341 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag 8401 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga 8461 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac 8521 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc 8581 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta 8641 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac 8701 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat 8761 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa 8821 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg 8881 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag 8941 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc 9001 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct 9061 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat 9121 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg 9181 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc 9241 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta 9301 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg 9361 tcgacggatc gggagatcaa cttgtttatt gcagcttata atggttacaa ataaagcaat 9421 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc 9481 aaactcatca atgtatctta tcatgtctgg atcaactgga taactcaagc taaccaaaat 9541 catcccaaac ttcccacccc ataccctatt accactgcca attacctgtg gtttcattta 9601 ctctaaacct gtgattcctc tgaattattt tcattttaaa gaaattgtat ttgttaaata 9661 tgtactacaa acttagtagt ttttaaagaa attgtatttg ttaaatatgt actacaaact 9721 tagtagt //