LOCUS V018334 8745 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V018334 VERSION V018334 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 2827..3471 /label="Hmgb1(NM_001313894)" /note="Hmgb1(NM_001313894)" /gene="Hmgb1" CDS 3478..3501 /codon_start=1 /product="FLAG® epitope tag, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDDDDK" /label="FLAG" promoter 3533..4032 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 4053..4652 /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 4666..5254 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5461..6094 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(6223..6239) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6247..6263 /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(6271..6301) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6316..6337 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6625..7213) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7384..8244) /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(8245..8349) /gene="bla" /label="AmpR promoter" polyA_signal 8397..8531 /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 gccaccatgg gcaaaggaga tcctaaaaag ccgagaggca aaatgtcctc atatgcattc 2881 tttgtgcaaa cttgccggga ggagcacaag aagaagcacc cggatgcttc tgtcaacttc 2941 tcagagttct ccaagaagtg ctcagagagg tggaagacca tgtctgctaa agaaaagggg 3001 aaatttgaag atatggcaaa ggctgacaag gctcgttatg aaagagaaat gaaaacctac 3061 atccccccca aaggggagac caaaaagaag ttcaaggacc ccaatgcacc caagaggcct 3121 ccttcggcct tcttcttgtt ctgttctgag taccgcccca aaatcaaagg cgagcatcct 3181 ggcttatcca ttggtgatgt tgcaaagaaa ctaggagaga tgtggaacaa cactgcagca 3241 gatgacaagc agccctatga gaagaaagct gccaagctga aggagaagta tgagaaggat 3301 attgctgcct acagagctaa aggaaaacct gatgcagcga aaaagggggt ggtcaaggct 3361 gaaaagagca agaaaaagaa ggaagaggaa gatgatgagg aggatgaaga ggatgaggaa 3421 gaggaggaag aagaggaaga cgaagatgaa gaagaagatg atgatgatga agaattcgat 3481 tacaaggatg acgacgataa gtgaggatcc cgcgactcta gataattcta ccgggtaggg 3541 gaggcgcttt tcccaaggca gtctggagca tgcgctttag cagccccgct gggcacttgg 3601 cgctacacaa gtggcctctg gcctcgcaca cattccacat ccaccggtag gcgccaaccg 3661 gctccgttct ttggtggccc cttcgcgcca ccttctactc ctcccctagt caggaagttc 3721 ccccccgccc cgcagctcgc gtcgtgcagg acgtgacaaa tggaagtagc acgtctcact 3781 agtctcgtgc agatggacag caccgctgag caatggaagc gggtaggcct ttggggcagc 3841 ggccaatagc agctttgctc cttcgctttc tgggctcaga ggctgggaag gggtgggtcc 3901 gggggcgggc tcaggggcgg gctcaggggc ggggcgggcg cccgaaggtc ctccggaggc 3961 ccggcattct gcacgcttca aaagcgcacg tctgccgcgc tgttctcctc ttcctcatct 4021 ccgggccttt cgacctgcag cccaagctta ccatgaccga gtacaagccc acggtgcgcc 4081 tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg ttcgccgact 4141 accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc accgagctgc 4201 aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg 4261 gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg 4321 agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg 4381 aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg 4441 tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg 4501 cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct 4561 tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca 4621 cctggtgcat gacccgcaag cccggtgcct gaccgcgtct ggaacaatca acctctggat 4681 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt 4741 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc 4801 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg 4861 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc 4921 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa 4981 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat 5041 tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg tgttgccacc 5101 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt 5161 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag 5221 acgagtcgga tctccctttg ggccgcctcc ccgcctggaa ttaattctgc agtcgagacc 5281 tagaaaaaca tggagcaatc acaagtagca atacagcagc taccaatgct gattgtgcct 5341 ggctagaagc acaagaggag gaggaggtgg gttttccagt cacacctcag gtacctttaa 5401 gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa aagaggggac 5461 tggaagggct aattcactcc caacgaagac aagatatcct tgatctgtgg atctaccaca 5521 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac 5581 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca 5641 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg 5701 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag 5761 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg 5821 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat 5881 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga 5941 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct 6001 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc 6061 agaccctttt agtcagtgtg gaaaatctct agcagtagta gttcatgtca tcttattatt 6121 cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggccttg acattgctag 6181 cgttttaccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt 6241 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa 6301 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc 6361 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag 6421 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt 6481 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga 6541 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg 6601 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa 6661 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt 6721 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct 6781 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct 6841 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc 6901 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt 6961 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc 7021 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat 7081 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa 7141 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa 7201 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga 7261 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct 7321 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga 7381 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc 7441 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg 7501 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat 7561 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat 7621 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg 7681 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc 7741 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa 7801 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc 7861 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt 7921 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag 7981 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt 8041 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag 8101 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac 8161 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc 8221 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca 8281 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg 8341 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatcaact 8401 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata 8461 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc 8521 atgtctggat caactggata actcaagcta accaaaatca tcccaaactt cccaccccat 8581 accctattac cactgccaat tacctgtggt ttcatttact ctaaacctgt gattcctctg 8641 aattattttc attttaaaga aattgtattt gttaaatatg tactacaaac ttagtagttt 8701 ttaaagaaat tgtatttgtt aaatatgtac tacaaactta gtagt //