LOCUS V045739 8704 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V045739 VERSION V045739 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..3391 /label="FTH1(NM_002032)" /note="FTH1(NM_002032)" /gene="FTH1" CDS 3407..3472 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3492..3991 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 4012..4611 /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 4625..5213 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5420..6053 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(6182..6198) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6206..6222 /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(6230..6260) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6275..6296 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6584..7172) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7343..8203) /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(8204..8308) /gene="bla" /label="AmpR promoter" polyA_signal 8356..8490 /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 ccatgacgac cgcgtccacc tcgcaggtgc gccagaacta 2881 ccaccaggac tcagaggccg ccatcaaccg ccagatcaac ctggagctct acgcctccta 2941 cgtttacctg tccatgtctt actactttga ccgcgatgat gtggctttga agaactttgc 3001 caaatacttt cttcaccaat ctcatgagga gagggaacat gctgagaaac tgatgaagct 3061 gcagaaccaa cgaggtggcc gaatcttcct tcaggatatc aagaaaccag actgtgatga 3121 ctgggagagc gggctgaatg caatggagtg tgcattacat ttggaaaaaa atgtgaatca 3181 gtcactactg gaactgcaca aactggccac tgacaaaaat gacccccatt tgtgtgactt 3241 cattgagaca cattacctga atgagcaggt gaaagccatc aaagaattgg gtgaccacgt 3301 gaccaacttg cgcaagatgg gagcgcccga atctggcttg gcggaatatc tctttgacaa 3361 gcacaccctg ggagacagtg ataatgaaag cggtggaggc ggatccgact acaaagacca 3421 tgacggtgat tataaagatc atgacatcga ctacaaggat gacgatgaca agtagtctag 3481 ataattctac cgggtagggg aggcgctttt cccaaggcag tctggagcat gcgctttagc 3541 agccccgctg ggcacttggc gctacacaag tggcctctgg cctcgcacac attccacatc 3601 caccggtagg cgccaaccgg ctccgttctt tggtggcccc ttcgcgccac cttctactcc 3661 tcccctagtc aggaagttcc cccccgcccc gcagctcgcg tcgtgcagga cgtgacaaat 3721 ggaagtagca cgtctcacta gtctcgtgca gatggacagc accgctgagc aatggaagcg 3781 ggtaggcctt tggggcagcg gccaatagca gctttgctcc ttcgctttct gggctcagag 3841 gctgggaagg ggtgggtccg ggggcgggct caggggcggg ctcaggggcg gggcgggcgc 3901 ccgaaggtcc tccggaggcc cggcattctg cacgcttcaa aagcgcacgt ctgccgcgct 3961 gttctcctct tcctcatctc cgggcctttc gacctgcagc ccaagcttac catgaccgag 4021 tacaagccca cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt acgcaccctc 4081 gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgatccgga ccgccacatc 4141 gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag 4201 gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa 4261 gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg 4321 gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg 4381 ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc 4441 gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc 4501 gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag 4561 gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg accgcgtctg 4621 gaacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt 4681 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc 4741 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga 4801 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc 4861 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct 4921 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg 4981 gctgttgggc actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct 5041 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc 5101 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg 5161 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggaat 5221 taattctgca gtcgagacct agaaaaacat ggagcaatca caagtagcaa tacagcagct 5281 accaatgctg attgtgcctg gctagaagca caagaggagg aggaggtggg ttttccagtc 5341 acacctcagg tacctttaag accaatgact tacaaggcag ctgtagatct tagccacttt 5401 ttaaaagaaa agaggggact ggaagggcta attcactccc aacgaagaca agatatcctt 5461 gatctgtgga tctaccacac acaaggctac ttccctgatt agcagaacta cacaccaggg 5521 ccaggggtca gatatccact gacctttgga tggtgctaca agctagtacc agttgagcca 5581 gataaggtag aagaggccaa taaaggagag aacaccagct tgttacaccc tgtgagcctg 5641 catgggatgg atgacccgga gagagaagtg ttagagtgga ggtttgacag ccgcctagca 5701 tttcatcacg tggcccgaga gctgcatccg gagtacttca agaactgctg atatcgagct 5761 tgctacaagg gactttccgc tggggacttt ccagggaggc gtggcctggg cgggactggg 5821 gagtggcgag ccctcagatc ctgcatataa gcagctgctt tttgcctgta ctgggtctct 5881 ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa 5941 gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc 6001 tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtagtag 6061 ttcatgtcat cttattattc agtatttata acttgcaaag aaatgaatat cagagagtga 6121 gaggccttga cattgctagc gttttaccgt cgacctctag ctagagcttg gcgtaatcat 6181 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag 6241 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg 6301 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa 6361 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca 6421 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg 6481 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc 6541 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc 6601 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac 6661 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc 6721 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata 6781 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc 6841 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca 6901 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag 6961 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta 7021 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg 7081 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc 7141 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt 7201 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa 7261 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat 7321 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga 7381 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac 7441 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg 7501 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg 7561 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt 7621 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct 7681 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat 7741 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta 7801 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca 7861 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat 7921 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac 7981 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa 8041 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt 8101 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg 8161 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat 8221 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt 8281 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg 8341 acggatcggg agatcaactt gtttattgca gcttataatg gttacaaata aagcaatagc 8401 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa 8461 ctcatcaatg tatcttatca tgtctggatc aactggataa ctcaagctaa ccaaaatcat 8521 cccaaacttc ccaccccata ccctattacc actgccaatt acctgtggtt tcatttactc 8581 taaacctgtg attcctctga attattttca ttttaaagaa attgtatttg ttaaatatgt 8641 actacaaact tagtagtttt taaagaaatt gtatttgtta aatatgtact acaaacttag 8701 tagt //