LOCUS V036718 9508 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V036718 VERSION V036718 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" CDS 2843..4177 /label="BCKDHA(NM_000709)" /note="BCKDHA(NM_000709)" /gene="BCKDHA" CDS 4184..4273 /codon_start=1 /product="3 tandem Myc epitope tags" /transl_table=1 /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL" /label="3xMyc" promoter 4296..4795 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 4816..5415 /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 5429..6017 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6224..6857 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(6986..7002) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7010..7026 /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(7034..7064) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7079..7100 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7388..7976) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8147..9007) /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(9008..9112) /gene="bla" /label="AmpR promoter" polyA_signal 9160..9294 /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 tcgctagcgc taccggactc agatctcgag 2821 ctcaagcttc gaattcgcca ccatggcggt agcgatcgct gcagcgaggg tctggcggct 2881 aaaccgtggt ttgagccagg ctgccctcct gctgctgcgg cagcctgggg ctcggggact 2941 ggctagatct caccccccca ggcagcagca gcagttttca tctctggatg acaagcccca 3001 gttcccaggg gcctcggcgg agtttataga taagttggaa ttcatccagc ccaacgtcat 3061 ctctggaatc cccatctacc gcgtcatgga ccggcaaggc cagatcatca accccagcga 3121 ggacccccac ctgccgaagg agaaggtgct gaagctctac aagagcatga cactgcttaa 3181 caccatggac cgcatcctct atgagtctca gcggcagggc cggatctcct tctacatgac 3241 caactatggt gaggagggca cgcacgtggg gagtgccgcc gccctggaca acacggacct 3301 ggtgtttggc cagtaccggg aggcaggtgt gctgatgtat cgggactacc ccctggaact 3361 attcatggcc cagtgctatg gcaacatcag tgacttgggc aaggggcgcc agatgcctgt 3421 ccactacggc tgcaaggaac gccacttcgt cactatctcc tctccactgg ccacgcagat 3481 ccctcaggcg gtgggggcgg cgtacgcagc caagcgggcc aatgccaaca gggtcgtcat 3541 ctgttacttc ggcgaggggg cagccagtga gggggacgcc catgccggct tcaacttcgc 3601 tgccacactt gagtgcccca tcatcttctt ctgccggaac aatggctacg ccatctccac 3661 gcccacctct gagcagtatc gcggcgatgg cattgcagca cgaggccccg ggtatggcat 3721 catgtcaatc cgcgtggatg gtaatgatgt gtttgccgta tacaacgcca caaaggaggc 3781 ccgacggcgg gctgtggcag agaaccagcc cttcctcatc gaggccatga cctacaggat 3841 cgggcaccac agcaccagtg acgacagttc agcgtaccgc tcggtggatg aggtcaatta 3901 ctgggataaa caggaccacc ccatctcccg gctgcggcac tatctgctga gccaaggctg 3961 gtgggatgag gagcaggaga aggcctggag gaagcagtcc cgcaggaagg tgatggaggc 4021 ctttgagcag gccgagcgga agcccaaacc caaccccaac ctactcttct cagacgtgta 4081 tcaggagatg cccgcccagc tccgcaagca gcaggagtct ctggcccgcc acctgcagac 4141 ctacggggag cactacccac tggatcactt cgataaggga tccgaacaaa aactaatatc 4201 agaggaagat ttggaacaga aactaatcag tgaagaagat ttagagcaaa agttaatttc 4261 cgaggaggac ttatagtgat ctagataatt ctaccgggta ggggaggcgc ttttcccaag 4321 gcagtctgga gcatgcgctt tagcagcccc gctgggcact tggcgctaca caagtggcct 4381 ctggcctcgc acacattcca catccaccgg taggcgccaa ccggctccgt tctttggtgg 4441 ccccttcgcg ccaccttcta ctcctcccct agtcaggaag ttcccccccg ccccgcagct 4501 cgcgtcgtgc aggacgtgac aaatggaagt agcacgtctc actagtctcg tgcagatgga 4561 cagcaccgct gagcaatgga agcgggtagg cctttggggc agcggccaat agcagctttg 4621 ctccttcgct ttctgggctc agaggctggg aaggggtggg tccgggggcg ggctcagggg 4681 cgggctcagg ggcggggcgg gcgcccgaag gtcctccgga ggcccggcat tctgcacgct 4741 tcaaaagcgc acgtctgccg cgctgttctc ctcttcctca tctccgggcc tttcgacctg 4801 cagcccaagc ttaccatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac 4861 gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac 4921 accgtcgatc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg 4981 cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc 5041 tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg 5101 gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct cctggcgccg 5161 caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag 5221 ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg 5281 gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc 5341 ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg catgacccgc 5401 aagcccggtg cctgaccgcg tctggaacaa tcaacctctg gattacaaaa tttgtgaaag 5461 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat 5521 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc 5581 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg 5641 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct 5701 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct 5761 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg 5821 gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac 5881 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct 5941 gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct 6001 ttgggccgcc tccccgcctg gaattaattc tgcagtcgag acctagaaaa acatggagca 6061 atcacaagta gcaatacagc agctaccaat gctgattgtg cctggctaga agcacaagag 6121 gaggaggagg tgggttttcc agtcacacct caggtacctt taagaccaat gacttacaag 6181 gcagctgtag atcttagcca ctttttaaaa gaaaagaggg gactggaagg gctaattcac 6241 tcccaacgaa gacaagatat ccttgatctg tggatctacc acacacaagg ctacttccct 6301 gattagcaga actacacacc agggccaggg gtcagatatc cactgacctt tggatggtgc 6361 tacaagctag taccagttga gccagataag gtagaagagg ccaataaagg agagaacacc 6421 agcttgttac accctgtgag cctgcatggg atggatgacc cggagagaga agtgttagag 6481 tggaggtttg acagccgcct agcatttcat cacgtggccc gagagctgca tccggagtac 6541 ttcaagaact gctgatatcg agcttgctac aagggacttt ccgctgggga ctttccaggg 6601 aggcgtggcc tgggcgggac tggggagtgg cgagccctca gatcctgcat ataagcagct 6661 gctttttgcc tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg 6721 ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt 6781 gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt 6841 gtggaaaatc tctagcagta gtagttcatg tcatcttatt attcagtatt tataacttgc 6901 aaagaaatga atatcagaga gtgagaggcc ttgacattgc tagcgtttta ccgtcgacct 6961 ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc 7021 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat 7081 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc 7141 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg 7201 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag 7261 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag 7321 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc 7381 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc 7441 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc 7501 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt 7561 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg 7621 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat 7681 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag 7741 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt 7801 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc 7861 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta 7921 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag 7981 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga 8041 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa 8101 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa 8161 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc 8221 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga 8281 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa 8341 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt 8401 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg 8461 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc 8521 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg 8581 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag 8641 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt 8701 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt 8761 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac 8821 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac 8881 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag 8941 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa 9001 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga 9061 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc 9121 cccgaaaagt gccacctgac gtcgacggat cgggagatca acttgtttat tgcagcttat 9181 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg 9241 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatcaactgg 9301 ataactcaag ctaaccaaaa tcatcccaaa cttcccaccc cataccctat taccactgcc 9361 aattacctgt ggtttcattt actctaaacc tgtgattcct ctgaattatt ttcattttaa 9421 agaaattgta tttgttaaat atgtactaca aacttagtag tttttaaaga aattgtattt 9481 gttaaatatg tactacaaac ttagtagt //