LOCUS V033147 10255 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V033147 VERSION V033147 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..4885 /label="ACOX2(NM_003500)" /note="ACOX2(NM_003500)" /gene="ACOX2" CDS 4892..4957 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" CDS 4958..5023 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 5043..5542 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 5563..6162 /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 6176..6764 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6971..7604 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(7733..7749) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7757..7773 /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(7781..7811) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7826..7847 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(8135..8723) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8894..9754) /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(9755..9859) /gene="bla" /label="AmpR promoter" polyA_signal 9907..10041 /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 ccatgggcag cccagtgcac cgagtgtcat tgggggatac 2881 ctggagcagg caaatgcacc ccgacataga gagcgagagg tatatgcagt cctttgacgt 2941 ggaacggctc accaacatcc ttgatggagg tgcccagaac actgcactcc gcaggaaagt 3001 tgagagcatc atccacagtt acccggagtt tagctgtaag gacaattatt tcatgaccca 3061 gaatgagcgt tataaggctg ccatgcggag ggcattccac atccggttga tagctcggcg 3121 cctgggttgg ttagaagatg gtcgtgaatt aggctacgct tacagagccc tttctggaga 3181 cgtggcctta aatatacaca gagtcttcgt gagagccctc aggagcctgg gctcagagga 3241 gcagattgcc aaatgggacc cactctgcaa aaacatccag atcatcgcaa cgtatgcaca 3301 gacagagttg ggacatggga catatcttca gggcctggag actgaagcca cctatgacgc 3361 agccacccag gagtttgtga tacacagccc cacgctgact gccaccaaat ggtggcctgg 3421 agacttggga cggtcagcca cccatgccct ggtccaggcc cagctgatct gctcaggagc 3481 caggcggggc atgcacgctt ttattgtgcc aatccggagt cttcaggacc acaccccact 3541 gccaggaatc atcattgggg acatcggacc caagatggac tttgatcaaa cagacaatgg 3601 cttcctgcag ctgaaccatg tgcgggtccc cagggagaac atgctgagtc gctttgcaca 3661 ggtcttgcca gatggcacct acgtcaaact cggtacagca cagagcaact accttcccat 3721 ggtggtggtg cgggtggagc tgctgtcagg ggagatcctc cctatactgc agaaggcctg 3781 tgtcatcgcc atgcgctact cggtcatccg ccgccaatcc cggctccggc ccagtgaccc 3841 agaggcaaag gtcctggact accagacaca acagcagaaa ctctttcctc agctggccat 3901 cagttatgcc ttccatttcc tggcagtcag cctcttggag ttcttccagc actcctacac 3961 tgccattctg aaccaagact tcagcttcct gcctgagctc cacgcactga gcacgggcat 4021 gaaggccatg atgtcagaat tctgcaccca gggagctgag atgtgccgca gggcctgtgg 4081 cggacatggc tactcaaagc tgagtggcct gccatcactg gtcaccaaat tgtcggcctc 4141 ctgtacctac gagggtgaga acacagtgct ctacctgcag gtggccaggt tcctggtgaa 4201 gagctacctg cagactcaga tgtcccctgg ctccacgcca cagagatctc tctctccatc 4261 tgtcgcatat ctcaccgcac ctgacctggc caggtgtcca gcccagaggg cagccgactt 4321 cctctgcccg gagctctaca ccacggcctg ggcacatgtg gcagtaaggc tcataaagga 4381 ctcagtgcag catttacaga ccctgacgca atccggagct gaccagcacg aggcttggaa 4441 ccagaccact gtcatacacc tccaggctgc taaggtgcac tgctactatg tcactgtgaa 4501 gggttttaca gaagctctgg agaaactaga aaatgaacca gcgattcagc aggtgctcaa 4561 gcgcctctgt gacctccatg ccatacatgg aatcttgact aactcgggtg actttctcca 4621 tgacgccttc ctgtctggtg cccaagtgga catggcaaga acagcctacc tggacctgct 4681 ccgcctgatc cggaaggatg ccatcctgtt aactgatgct tttgacttca ccgatcagtg 4741 tttaaattca gcacttggct gttatgatgg aaacgtctac gaacgcctgt tccagtgggc 4801 tcagaagtca ccaaccaata ctcaggagaa ccctgcctat gaggaatata taagaccact 4861 tttacaaagt tggagatcca agctaggatc cgactacaag gaccacgacg gcgactacaa 4921 ggaccacgac atcgactaca aggacgacga cgacaaggac tacaaagacc atgacggtga 4981 ttataaagat catgacatcg actacaagga tgacgatgac aagtagtcta gataattcta 5041 ccgggtaggg gaggcgcttt tcccaaggca gtctggagca tgcgctttag cagccccgct 5101 gggcacttgg cgctacacaa gtggcctctg gcctcgcaca cattccacat ccaccggtag 5161 gcgccaaccg gctccgttct ttggtggccc cttcgcgcca ccttctactc ctcccctagt 5221 caggaagttc ccccccgccc cgcagctcgc gtcgtgcagg acgtgacaaa tggaagtagc 5281 acgtctcact agtctcgtgc agatggacag caccgctgag caatggaagc gggtaggcct 5341 ttggggcagc ggccaatagc agctttgctc cttcgctttc tgggctcaga ggctgggaag 5401 gggtgggtcc gggggcgggc tcaggggcgg gctcaggggc ggggcgggcg cccgaaggtc 5461 ctccggaggc ccggcattct gcacgcttca aaagcgcacg tctgccgcgc tgttctcctc 5521 ttcctcatct ccgggccttt cgacctgcag cccaagctta ccatgaccga gtacaagccc 5581 acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg 5641 ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc 5701 accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc 5761 gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg 5821 gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag 5881 caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc 5941 accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc 6001 ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc 6061 aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa 6121 ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gaccgcgtct ggaacaatca 6181 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt 6241 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc 6301 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc 6361 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg 6421 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc 6481 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg 6541 cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg 6601 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc 6661 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct 6721 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggaa ttaattctgc 6781 agtcgagacc tagaaaaaca tggagcaatc acaagtagca atacagcagc taccaatgct 6841 gattgtgcct ggctagaagc acaagaggag gaggaggtgg gttttccagt cacacctcag 6901 gtacctttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa 6961 aagaggggac tggaagggct aattcactcc caacgaagac aagatatcct tgatctgtgg 7021 atctaccaca cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc 7081 agatatccac tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta 7141 gaagaggcca ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg 7201 gatgacccgg agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac 7261 gtggcccgag agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag 7321 ggactttccg ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga 7381 gccctcagat cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga 7441 ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata 7501 aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta 7561 gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtagta gttcatgtca 7621 tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggccttg 7681 acattgctag cgttttaccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc 7741 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca 7801 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct 7861 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac 7921 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc 7981 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt 8041 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg 8101 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg 8161 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat 8221 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta 8281 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct 8341 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc 8401 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa 8461 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg 8521 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag 8581 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt 8641 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta 8701 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc 8761 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca 8821 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa 8881 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat 8941 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct 9001 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt 9061 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat 9121 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta 9181 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg 9241 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt 9301 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg 9361 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg 9421 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc 9481 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa 9541 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac 9601 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt 9661 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg 9721 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa 9781 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata 9841 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg 9901 gagatcaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat 9961 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat 10021 gtatcttatc atgtctggat caactggata actcaagcta accaaaatca tcccaaactt 10081 cccaccccat accctattac cactgccaat tacctgtggt ttcatttact ctaaacctgt 10141 gattcctctg aattattttc attttaaaga aattgtattt gttaaatatg tactacaaac 10201 ttagtagttt ttaaagaaat tgtatttgtt aaatatgtac tacaaactta gtagt //