LOCUS V032367 10174 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V032367 VERSION V032367 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..4870 /label="ALOX15B(NM_001141)" /note="ALOX15B(NM_001141)" /gene="ALOX15B" CDS 4877..4942 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 4962..5461 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 5482..6081 /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 6095..6683 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6890..7523 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(7652..7668) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7676..7692 /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(7700..7730) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7745..7766 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(8054..8642) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8813..9673) /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(9674..9778) /gene="bla" /label="AmpR promoter" polyA_signal 9826..9960 /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 ccatggccga gttcagggtc agggtgtcca ccggagaagc 2881 cttcggggct ggcacatggg acaaagtgtc tgtcagcatc gtggggaccc ggggagagag 2941 ccccccactg cccctggaca atctcggcaa ggagttcact gcgggcgctg aggaggactt 3001 ccaggtgacg ctcccggagg acgtaggccg agtgctgctg ctgcgcgtgc acaaggcgcc 3061 cccagtgctg cccctgctgg ggcccctggc cccggatgcc tggttctgcc gctggttcca 3121 gctgacaccg ccgcggggcg gccacctcct cttcccctgc taccagtggc tggagggggc 3181 ggggaccctg gtgctgcagg agggtacagc caaggtgtcc tgggcagacc accaccctgt 3241 gctccagcaa cagcgccagg aggagcttca ggcccggcag gagatgtacc agtggaaggc 3301 ttacaaccca ggttggcctc actgcctgga tgaaaagaca gtggaagact tggagctcaa 3361 tatcaaatac tccacagcca agaatgccaa cttttatcta caggctggct ctgcttttgc 3421 agagatgaaa atcaaggggt tgctggaccg caaggggctc tggaggagtc tgaatgagat 3481 gaaaaggatc ttcaacttcc ggaggacccc agcagctgag cacgcatttg agcactggca 3541 ggaggacgcc ttcttcgcct cccagttcct gaatggtctc aaccctgtcc tgatccgccg 3601 ctgtcactac ctcccaaaga acttccccgt cactgatgcc atggtggcct cagtgttggg 3661 tcctgggacc agcttgcagg ctgagctaga gaagggctcc ctgttcttgg tggatcacgg 3721 catcctctct ggcatccaga ccaatgtcat taatgggaag cctcagttct ctgcggcccc 3781 aatgaccctg ctataccaga gcccaggctg cgggccgctg ctgcctctcg ccatccagct 3841 cagccagacc cccggcccaa acagccccat cttcctgccc actgatgaca agtgggactg 3901 gttgctggcc aagacctggg tgcgcaatgc cgagttctcc ttccatgagg ccctcacgca 3961 cctgctgcac tcacatctgc tgcctgaggt cttcaccctg gctaccctgc gtcagctgcc 4021 ccactgccac cctctcttca agctgctgat cccgcacacc cgatacaccc tgcacatcaa 4081 cacactcgcc cgggagctgc ttatcgtgcc agggcaggtg gtggacaggt ccacaggcat 4141 cggcattgaa ggcttctctg agttgataca gaggaacatg aagcagctga actattctct 4201 cctgtgtctg cctgaggata tccggacccg aggagttgaa gacatcccag gctactacta 4261 ccgtgatgat gggatgcaga tctggggtgc agtggaacgc tttgtctctg aaatcatcgg 4321 tatctactac ccaagtgatg agtctgtcca agatgacaga gagctccagg cctgggtcag 4381 agagatcttc tccaagggct tcctaaacca ggagagctca ggtataccct cctcactgga 4441 gacccgggaa gccctggtgc agtatgtcac catggtgata ttcacctgct ccgccaagca 4501 tgcggctgtc agtgcagggc agtttgactc ctgtgcttgg atgcccaacc tgccacccag 4561 catgcagctg ccaccaccca cctccaaagg cctggcaaca tgcgagggct tcatagccac 4621 cctcccacct gtcaatgcca catgtgatgt catccttgct ctctggttgc tgagcaagga 4681 gcctggagac caaaggcccc tgggcaccta tccggatgag cacttcacag aggaggcccc 4741 tcggcggagc atcgccacct tccagagccg cctggcccag atctcgaggg gcatccagga 4801 gcggaaccag ggcctggtgc tgccctacac ctacctagac cctcccctca tcgagaacag 4861 cgtctccatc ggatccgact acaaagacca tgacggtgat tataaagatc atgacatcga 4921 ctacaaggat gacgatgaca agtagtctag ataattctac cgggtagggg aggcgctttt 4981 cccaaggcag tctggagcat gcgctttagc agccccgctg ggcacttggc gctacacaag 5041 tggcctctgg cctcgcacac attccacatc caccggtagg cgccaaccgg ctccgttctt 5101 tggtggcccc ttcgcgccac cttctactcc tcccctagtc aggaagttcc cccccgcccc 5161 gcagctcgcg tcgtgcagga cgtgacaaat ggaagtagca cgtctcacta gtctcgtgca 5221 gatggacagc accgctgagc aatggaagcg ggtaggcctt tggggcagcg gccaatagca 5281 gctttgctcc ttcgctttct gggctcagag gctgggaagg ggtgggtccg ggggcgggct 5341 caggggcggg ctcaggggcg gggcgggcgc ccgaaggtcc tccggaggcc cggcattctg 5401 cacgcttcaa aagcgcacgt ctgccgcgct gttctcctct tcctcatctc cgggcctttc 5461 gacctgcagc ccaagcttac catgaccgag tacaagccca cggtgcgcct cgccacccgc 5521 gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg 5581 cgccacaccg tcgatccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc 5641 ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg 5701 gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg 5761 cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga aggcctcctg 5821 gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac 5881 caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc 5941 gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg 6001 ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg 6061 acccgcaagc ccggtgcctg accgcgtctg gaacaatcaa cctctggatt acaaaatttg 6121 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc 6181 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta 6241 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt 6301 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca 6361 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc 6421 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt 6481 gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct ggattctgcg 6541 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg 6601 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat 6661 ctccctttgg gccgcctccc cgcctggaat taattctgca gtcgagacct agaaaaacat 6721 ggagcaatca caagtagcaa tacagcagct accaatgctg attgtgcctg gctagaagca 6781 caagaggagg aggaggtggg ttttccagtc acacctcagg tacctttaag accaatgact 6841 tacaaggcag ctgtagatct tagccacttt ttaaaagaaa agaggggact ggaagggcta 6901 attcactccc aacgaagaca agatatcctt gatctgtgga tctaccacac acaaggctac 6961 ttccctgatt agcagaacta cacaccaggg ccaggggtca gatatccact gacctttgga 7021 tggtgctaca agctagtacc agttgagcca gataaggtag aagaggccaa taaaggagag 7081 aacaccagct tgttacaccc tgtgagcctg catgggatgg atgacccgga gagagaagtg 7141 ttagagtgga ggtttgacag ccgcctagca tttcatcacg tggcccgaga gctgcatccg 7201 gagtacttca agaactgctg atatcgagct tgctacaagg gactttccgc tggggacttt 7261 ccagggaggc gtggcctggg cgggactggg gagtggcgag ccctcagatc ctgcatataa 7321 gcagctgctt tttgcctgta ctgggtctct ctggttagac cagatctgag cctgggagct 7381 ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca 7441 agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta 7501 gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat cttattattc agtatttata 7561 acttgcaaag aaatgaatat cagagagtga gaggccttga cattgctagc gttttaccgt 7621 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt 7681 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg 7741 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg 7801 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc 7861 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc 7921 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata 7981 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg 8041 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct 8101 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa 8161 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc 8221 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt 8281 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg 8341 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg 8401 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct 8461 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc 8521 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg 8581 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc 8641 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt 8701 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa 8761 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat 8821 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct 8881 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg 8941 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag 9001 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta 9061 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg 9121 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg 9181 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct 9241 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta 9301 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg 9361 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc 9421 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg 9481 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga 9541 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg 9601 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat 9661 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc 9721 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca 9781 catttccccg aaaagtgcca cctgacgtcg acggatcggg agatcaactt gtttattgca 9841 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt 9901 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc 9961 aactggataa ctcaagctaa ccaaaatcat cccaaacttc ccaccccata ccctattacc 10021 actgccaatt acctgtggtt tcatttactc taaacctgtg attcctctga attattttca 10081 ttttaaagaa attgtatttg ttaaatatgt actacaaact tagtagtttt taaagaaatt 10141 gtatttgtta aatatgtact acaaacttag tagt //