LOCUS V023999 8884 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V023999 VERSION V023999 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..3580 /label="TFAM(NM_003201)" /note="TFAM(NM_003201)" /gene="TFAM" CDS 3587..3652 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3672..4171 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 4192..4791 /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 4805..5393 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5600..6233 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(6362..6378) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6386..6402 /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(6410..6440) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6455..6476 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6764..7352) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7523..8383) /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(8384..8488) /gene="bla" /label="AmpR promoter" polyA_signal 8536..8670 /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 ccatggcgtt tctccgaagc atgtggggcg tgctgagtgc 2881 cctgggaagg tctggagcag agctgtgcac cggctgtgga agtcgactgc gctccccctt 2941 cagttttgtg tatttaccga ggtggttttc atctgtcttg gcaagttgtc caaagaaacc 3001 tgtaagttct taccttcgat tttctaaaga acaactaccc atatttaaag ctcagaaccc 3061 agatgcaaaa actacagaac taattagaag aattgcccag cgttggaggg aacttcctga 3121 ttcaaagaaa aaaatatatc aagatgctta tagggcggag tggcaggtat ataaagaaga 3181 gataagcaga tttaaagaac agctaactcc aagtcagatt atgtctttgg aaaaagaaat 3241 catggacaaa catttaaaaa ggaaagctat gacaaaaaaa aaagagttaa cactgcttgg 3301 aaaaccaaaa agacctcgtt cagcttataa cgtttatgta gctgaaagat tccaagaagc 3361 taagggtgat tcaccgcagg aaaagctgaa gactgtaaag gaaaactgga aaaatctgtc 3421 tgactctgaa aaggaattat atattcagca tgctaaagag gacgaaactc gttatcataa 3481 tgaaatgaag tcttgggaag aacaaatgat tgaagttgga cgaaaggatc ttctacgtcg 3541 cacaataaag aaacaacgaa aatatggtgc tgaggagtgt ggatccgact acaaagacca 3601 tgacggtgat tataaagatc atgacatcga ctacaaggat gacgatgaca agtagtctag 3661 ataattctac cgggtagggg aggcgctttt cccaaggcag tctggagcat gcgctttagc 3721 agccccgctg ggcacttggc gctacacaag tggcctctgg cctcgcacac attccacatc 3781 caccggtagg cgccaaccgg ctccgttctt tggtggcccc ttcgcgccac cttctactcc 3841 tcccctagtc aggaagttcc cccccgcccc gcagctcgcg tcgtgcagga cgtgacaaat 3901 ggaagtagca cgtctcacta gtctcgtgca gatggacagc accgctgagc aatggaagcg 3961 ggtaggcctt tggggcagcg gccaatagca gctttgctcc ttcgctttct gggctcagag 4021 gctgggaagg ggtgggtccg ggggcgggct caggggcggg ctcaggggcg gggcgggcgc 4081 ccgaaggtcc tccggaggcc cggcattctg cacgcttcaa aagcgcacgt ctgccgcgct 4141 gttctcctct tcctcatctc cgggcctttc gacctgcagc ccaagcttac catgaccgag 4201 tacaagccca cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt acgcaccctc 4261 gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgatccgga ccgccacatc 4321 gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag 4381 gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa 4441 gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg 4501 gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg 4561 ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc 4621 gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc 4681 gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag 4741 gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg accgcgtctg 4801 gaacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt 4861 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc 4921 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga 4981 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc 5041 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct 5101 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg 5161 gctgttgggc actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct 5221 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc 5281 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg 5341 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggaat 5401 taattctgca gtcgagacct agaaaaacat ggagcaatca caagtagcaa tacagcagct 5461 accaatgctg attgtgcctg gctagaagca caagaggagg aggaggtggg ttttccagtc 5521 acacctcagg tacctttaag accaatgact tacaaggcag ctgtagatct tagccacttt 5581 ttaaaagaaa agaggggact ggaagggcta attcactccc aacgaagaca agatatcctt 5641 gatctgtgga tctaccacac acaaggctac ttccctgatt agcagaacta cacaccaggg 5701 ccaggggtca gatatccact gacctttgga tggtgctaca agctagtacc agttgagcca 5761 gataaggtag aagaggccaa taaaggagag aacaccagct tgttacaccc tgtgagcctg 5821 catgggatgg atgacccgga gagagaagtg ttagagtgga ggtttgacag ccgcctagca 5881 tttcatcacg tggcccgaga gctgcatccg gagtacttca agaactgctg atatcgagct 5941 tgctacaagg gactttccgc tggggacttt ccagggaggc gtggcctggg cgggactggg 6001 gagtggcgag ccctcagatc ctgcatataa gcagctgctt tttgcctgta ctgggtctct 6061 ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa 6121 gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc 6181 tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtagtag 6241 ttcatgtcat cttattattc agtatttata acttgcaaag aaatgaatat cagagagtga 6301 gaggccttga cattgctagc gttttaccgt cgacctctag ctagagcttg gcgtaatcat 6361 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag 6421 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg 6481 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa 6541 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca 6601 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg 6661 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc 6721 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc 6781 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac 6841 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc 6901 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata 6961 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc 7021 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca 7081 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag 7141 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta 7201 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg 7261 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc 7321 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt 7381 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa 7441 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat 7501 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga 7561 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac 7621 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg 7681 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg 7741 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt 7801 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct 7861 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat 7921 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta 7981 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca 8041 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat 8101 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac 8161 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa 8221 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt 8281 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg 8341 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat 8401 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt 8461 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg 8521 acggatcggg agatcaactt gtttattgca gcttataatg gttacaaata aagcaatagc 8581 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa 8641 ctcatcaatg tatcttatca tgtctggatc aactggataa ctcaagctaa ccaaaatcat 8701 cccaaacttc ccaccccata ccctattacc actgccaatt acctgtggtt tcatttactc 8761 taaacctgtg attcctctga attattttca ttttaaagaa attgtatttg ttaaatatgt 8821 actacaaact tagtagtttt taaagaaatt gtatttgtta aatatgtact acaaacttag 8881 tagt //