LOCUS V045421 9970 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V045421 VERSION V045421 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..4657 /label="NFE2L2(NM_006164)" /note="NFE2L2(NM_006164)" /gene="NFE2L2" CDS 4673..4738 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 4758..5257 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 5278..5877 /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 5891..6479 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6686..7319 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(7448..7464) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7472..7488 /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(7496..7526) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7541..7562 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7850..8438) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8609..9469) /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(9470..9574) /gene="bla" /label="AmpR promoter" polyA_signal 9622..9756 /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 ccatgatgga cttggagctg ccgccgccgg gactcccgtc 2881 ccagcaggac atggatttga ttgacatact ttggaggcaa gatatagatc ttggagtaag 2941 tcgagaagta tttgacttca gtcagcgacg gaaagagtat gagctggaaa aacagaaaaa 3001 acttgaaaag gaaagacaag aacaactcca aaaggagcaa gagaaagcct ttttcgctca 3061 gttacaacta gatgaagaga caggtgaatt tctcccaatt cagccagccc agcacatcca 3121 gtcagaaacc agtggatctg ccaactactc ccaggttgcc cacattccca aatcagatgc 3181 tttgtacttt gatgactgca tgcagctttt ggcgcagaca ttcccgtttg tagatgacaa 3241 tgaggtttct tcggctacgt ttcagtcact tgttcctgat attcccggtc acatcgagag 3301 cccagtcttc attgctacta atcaggctca gtcacctgaa acttctgttg ctcaggtagc 3361 ccctgttgat ttagacggta tgcaacagga cattgagcaa gtttgggagg agctattatc 3421 cattcctgag ttacagtgtc ttaatattga aaatgacaag ctggttgaga ctaccatggt 3481 tccaagtcca gaagccaaac tgacagaagt tgacaattat catttttact catctatacc 3541 ctcaatggaa aaagaagtag gtaactgtag tccacatttt cttaatgctt ttgaggattc 3601 cttcagcagc atcctctcca cagaagaccc caaccagttg acagtgaact cattaaattc 3661 agatgccaca gtcaacacag attttggtga tgaattttat tctgctttca tagctgagcc 3721 cagtatcagc aacagcatgc cctcacctgc tactttaagc cattcactct ctgaacttct 3781 aaatgggccc attgatgttt ctgatctatc actttgcaaa gctttcaacc aaaaccaccc 3841 tgaaagcaca gcagaattca atgattctga ctccggcatt tcactaaaca caagtcccag 3901 tgtggcatca ccagaacact cagtggaatc ttccagctat ggagacacac tacttggcct 3961 cagtgattct gaagtggaag agctagatag tgcccctgga agtgtcaaac agaatggtcc 4021 taaaacacca gtacattctt ctggggatat ggtacaaccc ttgtcaccat ctcaggggca 4081 gagcactcac gtgcatgatg cccaatgtga gaacacacca gagaaagaat tgcctgtaag 4141 tcctggtcat cggaaaaccc cattcacaaa agacaaacat tcaagccgct tggaggctca 4201 tctcacaaga gatgaactta gggcaaaagc tctccatatc ccattccctg tagaaaaaat 4261 cattaacctc cctgttgttg acttcaacga aatgatgtcc aaagagcagt tcaatgaagc 4321 tcaacttgca ttaattcggg atatacgtag gaggggtaag aataaagtgg ctgctcagaa 4381 ttgcagaaaa agaaaactgg aaaatatagt agaactagag caagatttag atcatttgaa 4441 agatgaaaaa gaaaaattgc tcaaagaaaa aggagaaaat gacaaaagcc ttcacctact 4501 gaaaaaacaa ctcagcacct tatatctcga agttttcagc atgctacgtg atgaagatgg 4561 aaaaccttat tctcctagtg aatactccct gcagcaaaca agagatggca atgttttcct 4621 tgttcccaaa agtaagaagc cagatgttaa gaaaaacggt ggaggcggat ccgactacaa 4681 agaccatgac ggtgattata aagatcatga catcgactac aaggatgacg atgacaagta 4741 gtctagataa ttctaccggg taggggaggc gcttttccca aggcagtctg gagcatgcgc 4801 tttagcagcc ccgctgggca cttggcgcta cacaagtggc ctctggcctc gcacacattc 4861 cacatccacc ggtaggcgcc aaccggctcc gttctttggt ggccccttcg cgccaccttc 4921 tactcctccc ctagtcagga agttcccccc cgccccgcag ctcgcgtcgt gcaggacgtg 4981 acaaatggaa gtagcacgtc tcactagtct cgtgcagatg gacagcaccg ctgagcaatg 5041 gaagcgggta ggcctttggg gcagcggcca atagcagctt tgctccttcg ctttctgggc 5101 tcagaggctg ggaaggggtg ggtccggggg cgggctcagg ggcgggctca ggggcggggc 5161 gggcgcccga aggtcctccg gaggcccggc attctgcacg cttcaaaagc gcacgtctgc 5221 cgcgctgttc tcctcttcct catctccggg cctttcgacc tgcagcccaa gcttaccatg 5281 accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc 5341 accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga tccggaccgc 5401 cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc 5461 ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc 5521 gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc 5581 cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc caaggagccc 5641 gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc agggcaaggg tctgggcagc 5701 gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag 5761 acctccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac 5821 gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg tgcctgaccg 5881 cgtctggaac aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa 5941 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat 6001 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta 6061 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc 6121 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt 6181 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg 6241 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc 6301 atggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc 6361 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct 6421 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc 6481 tggaattaat tctgcagtcg agacctagaa aaacatggag caatcacaag tagcaataca 6541 gcagctacca atgctgattg tgcctggcta gaagcacaag aggaggagga ggtgggtttt 6601 ccagtcacac ctcaggtacc tttaagacca atgacttaca aggcagctgt agatcttagc 6661 cactttttaa aagaaaagag gggactggaa gggctaattc actcccaacg aagacaagat 6721 atccttgatc tgtggatcta ccacacacaa ggctacttcc ctgattagca gaactacaca 6781 ccagggccag gggtcagata tccactgacc tttggatggt gctacaagct agtaccagtt 6841 gagccagata aggtagaaga ggccaataaa ggagagaaca ccagcttgtt acaccctgtg 6901 agcctgcatg ggatggatga cccggagaga gaagtgttag agtggaggtt tgacagccgc 6961 ctagcatttc atcacgtggc ccgagagctg catccggagt acttcaagaa ctgctgatat 7021 cgagcttgct acaagggact ttccgctggg gactttccag ggaggcgtgg cctgggcggg 7081 actggggagt ggcgagccct cagatcctgc atataagcag ctgctttttg cctgtactgg 7141 gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact 7201 gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg 7261 tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag 7321 tagtagttca tgtcatctta ttattcagta tttataactt gcaaagaaat gaatatcaga 7381 gagtgagagg ccttgacatt gctagcgttt taccgtcgac ctctagctag agcttggcgt 7441 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca 7501 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat 7561 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt 7621 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct 7681 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa 7741 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa 7801 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc 7861 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga 7921 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc 7981 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt 8041 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct 8101 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg 8161 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta 8221 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct 8281 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa 8341 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt 8401 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta 8461 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat 8521 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa 8581 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct 8641 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta 8701 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct 8761 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg 8821 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa 8881 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt 8941 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta 9001 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca 9061 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta 9121 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct 9181 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg 9241 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac 9301 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact 9361 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa 9421 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt 9481 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat 9541 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg 9601 acgtcgacgg atcgggagat caacttgttt attgcagctt ataatggtta caaataaagc 9661 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg 9721 tccaaactca tcaatgtatc ttatcatgtc tggatcaact ggataactca agctaaccaa 9781 aatcatccca aacttcccac cccataccct attaccactg ccaattacct gtggtttcat 9841 ttactctaaa cctgtgattc ctctgaatta ttttcatttt aaagaaattg tatttgttaa 9901 atatgtacta caaacttagt agtttttaaa gaaattgtat ttgttaaata tgtactacaa 9961 acttagtagt //