LOCUS V020808 8744 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V020808 VERSION V020808 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 2826..3440 /label="ATP6V0B(NM_004047)" /note="ATP6V0B(NM_004047)" /gene="ATP6V0B" CDS 3447..3512 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3532..4031 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 4052..4651 /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 4665..5253 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5460..6093 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(6222..6238) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6246..6262 /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(6270..6300) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6315..6336 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6624..7212) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7383..8243) /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(8244..8348) /gene="bla" /label="AmpR promoter" polyA_signal 8396..8530 /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 ccaccatgac ggggctagca ctgctctact ccggggtctt cgtggccttc tgggcctgcg 2881 cgctggccgt gggagtctgc tacaccattt ttgatttggg cttccgcttt gatgtggcat 2941 ggttcctgac ggagacttcg cccttcatgt ggtccaacct gggcattggc ctagctatct 3001 ccctgtctgt ggttggggca gcctggggca tctatattac cggctcctcc atcattggtg 3061 gaggagtgaa ggcccccagg atcaagacca agaacctggt cagcatcatc ttctgtgagg 3121 ctgtggccat ctacggcatc atcatggcaa ttgtcattag caacatggct gagcctttca 3181 gtgccacaga ccccaaggcc atcggccatc ggaactacca tgcaggctac tccatgtttg 3241 gggctggcct caccgtaggc ctgtctaacc tcttctgtgg agtctgcgtg ggcatcgtgg 3301 gcagtggggc tgccctggcc gatgctcaga accccagcct ctttgtaaag attctcatcg 3361 tggagatctt tggcagcgcc attggcctct ttggggtcat cgtcgcaatt cttcagacct 3421 ccagagtgaa gatgggtgac ggatccgact acaaagacca tgacggtgat tataaagatc 3481 atgacatcga ctacaaggat gacgatgaca agtaatctag ataattctac cgggtagggg 3541 aggcgctttt cccaaggcag tctggagcat gcgctttagc agccccgctg ggcacttggc 3601 gctacacaag tggcctctgg cctcgcacac attccacatc caccggtagg cgccaaccgg 3661 ctccgttctt tggtggcccc ttcgcgccac cttctactcc tcccctagtc aggaagttcc 3721 cccccgcccc gcagctcgcg tcgtgcagga cgtgacaaat ggaagtagca cgtctcacta 3781 gtctcgtgca gatggacagc accgctgagc aatggaagcg ggtaggcctt tggggcagcg 3841 gccaatagca gctttgctcc ttcgctttct gggctcagag gctgggaagg ggtgggtccg 3901 ggggcgggct caggggcggg ctcaggggcg gggcgggcgc ccgaaggtcc tccggaggcc 3961 cggcattctg cacgcttcaa aagcgcacgt ctgccgcgct gttctcctct tcctcatctc 4021 cgggcctttc gacctgcagc ccaagcttac catgaccgag tacaagccca cggtgcgcct 4081 cgccacccgc gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt tcgccgacta 4141 ccccgccacg cgccacaccg tcgatccgga ccgccacatc gagcgggtca ccgagctgca 4201 agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg 4261 cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga 4321 gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga 4381 aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt 4441 ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc 4501 ggccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt 4561 ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac 4621 ctggtgcatg acccgcaagc ccggtgcctg accgcgtctg gaacaatcaa cctctggatt 4681 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg 4741 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct 4801 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc 4861 aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca 4921 ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac 4981 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt 5041 ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct 5101 ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc 5161 cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga 5221 cgagtcggat ctccctttgg gccgcctccc cgcctggaat taattctgca gtcgagacct 5281 agaaaaacat ggagcaatca caagtagcaa tacagcagct accaatgctg attgtgcctg 5341 gctagaagca caagaggagg aggaggtggg ttttccagtc acacctcagg tacctttaag 5401 accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa agaggggact 5461 ggaagggcta attcactccc aacgaagaca agatatcctt gatctgtgga tctaccacac 5521 acaaggctac ttccctgatt agcagaacta cacaccaggg ccaggggtca gatatccact 5581 gacctttgga tggtgctaca agctagtacc agttgagcca gataaggtag aagaggccaa 5641 taaaggagag aacaccagct tgttacaccc tgtgagcctg catgggatgg atgacccgga 5701 gagagaagtg ttagagtgga ggtttgacag ccgcctagca tttcatcacg tggcccgaga 5761 gctgcatccg gagtacttca agaactgctg atatcgagct tgctacaagg gactttccgc 5821 tggggacttt ccagggaggc gtggcctggg cgggactggg gagtggcgag ccctcagatc 5881 ctgcatataa gcagctgctt tttgcctgta ctgggtctct ctggttagac cagatctgag 5941 cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt 6001 gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca 6061 gaccctttta gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat cttattattc 6121 agtatttata acttgcaaag aaatgaatat cagagagtga gaggccttga cattgctagc 6181 gttttaccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg 6241 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa 6301 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct 6361 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga 6421 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc 6481 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa 6541 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt 6601 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa 6661 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt 6721 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg 6781 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc 6841 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc 6901 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta 6961 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct 7021 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc 7081 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa 7141 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa 7201 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa 7261 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt 7321 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac 7381 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc 7441 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc 7501 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata 7561 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc 7621 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc 7681 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca 7741 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa 7801 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca 7861 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt 7921 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt 7981 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg 8041 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga 8101 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc 8161 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg 8221 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag 8281 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg 8341 gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg agatcaactt 8401 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa 8461 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca 8521 tgtctggatc aactggataa ctcaagctaa ccaaaatcat cccaaacttc ccaccccata 8581 ccctattacc actgccaatt acctgtggtt tcatttactc taaacctgtg attcctctga 8641 attattttca ttttaaagaa attgtatttg ttaaatatgt actacaaact tagtagtttt 8701 taaagaaatt gtatttgtta aatatgtact acaaacttag tagt //