LOCUS V023587 11028 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V023587 VERSION V023587 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 2815..5640 /label="USP33(NM_015017)" /note="USP33(NM_015017)" /gene="USP33" CDS 5647..5712 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 5731..6317 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" /label="IRES2" CDS 6337..6936 /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 6950..7538 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 7745..8378 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(8506..8522) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 8530..8546 /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(8554..8584) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 8599..8620 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(8908..9496) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(9667..10527) /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(10528..10632) /gene="bla" /label="AmpR promoter" polyA_signal 10680..10814 /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 tctatttccg gtgaattcgc caccatgaca 2821 ggatcaaatt cacacataac gatattaacc ttaaaggtgt tacctcattt tgaaagtctt 2881 gggaaacagg aaaaaattcc taacaaaatg tcagcttttc gaaatcattg tccacatttg 2941 gattcagttg gtgaaataac aaaagaagat ttgatacaaa aatcccttgg tacttgtcag 3001 gattgtaaag tccaaggacc aaatctttgg gcatgtctgg agaatagatg ttcatatgtt 3061 ggctgtggtg aatcacaagt agatcacagc accatacatt ctcaggagac aaagcattat 3121 ctaactgtga accttaccac tcttcgagta tggtgttatg cttgcagcaa agaagtattt 3181 ttggatagga aattaggaac tcagccttca ttgcctcatg taagacaacc tcaccaaata 3241 caagaaaaca gtgtccagga ttttaaaata cccagtaata caacattaaa aactcctctg 3301 gttgccgtat ttgatgatct ggatatagaa gcggatgaag aagatgaact tagggccaga 3361 ggtcttacag gtttgaaaaa tattggaaat acttgttaca tgaatgcagc tttgcaggct 3421 ctttctaatt gcccaccttt gacacagttt tttcttgatt gtggaggact agctcgaaca 3481 gataagaaac ctgccatttg taaaagttat ctcaaactaa tgacagagct gtggcataaa 3541 agcaggccag gatctgttgt gcctactact ctgtttcaag gaattaaaac tgtaaatcca 3601 acatttcggg ggtattctca gcaggatgct caagaattcc ttcgatgttt aatggatttg 3661 cttcatgaag aattgaaaga gcaagtcatg gaagtagaag aagatccgca aaccataacc 3721 actgaggaga caatggaaga agacaagagc cagtcggatg tagattttca gtcttgtgaa 3781 tcttgtagca acagtgatag agcagaaaat gaaaatggct ctagatgctt ttctgaagat 3841 aataatgaaa caacaatgtt aattcaggat gatgaaaaca attcagaaat gtcaaaggat 3901 tggcaaaaag agaagatgtg caataagatt aataaagtaa attctgaagg cgaatttgat 3961 aaagatagag actctatatc tgaaacagtc gacttaaaca accaggaaac tgtcaaagtg 4021 caaatacaca gcagagcttc agaatatatc actgatgtcc attcgaatga cctgtctaca 4081 ccacagatcc ttccatcaaa tgaaggtgtt aatccacgtt tatcggcaag ccctcctaaa 4141 tcaggcaatt tgtggccagg attggcacca ccacacaaaa aagctcagtc tgcatctcca 4201 aagagaaaaa aacagcacaa gaaatacaga agtgttattt cagacatatt tgatggaaca 4261 atcattagtt cagtgcagtg tctgacttgt gacagggtgt ctgtaaccct cgagaccttt 4321 caagatctgt ccttgccaat tcctggcaag gaagaccttg ctaagctgca ttcatcaagt 4381 catccaactt ctatagtcaa agcaggatca tgtggcgaag catatgctcc acaagggtgg 4441 atagcttttt tcatggaata tgtgaagagg tttgttgtct catgtgtccc tagctggttt 4501 tggggtccag tagtaacctt gcaagattgt cttgctgcct tctttgccag agatgaacta 4561 aaaggtgaca atatgtacag ttgtgaaaaa tgcaaaaagt tgagaaatgg agtgaagttt 4621 tgtaaagtac aaaactttcc tgagattttg tgcatccacc ttaaaagatt cagacatgaa 4681 ctaatgtttt ccaccaaaat cagtacccat gtttcatttc cgctagaagg cttggatctt 4741 cagccatttc ttgctaagga tagtccagct caaattgtga catatgatct tctgtcagtc 4801 atttgccatc atggaactgc aagtagtgga cactatatag cctactgccg aaacaatcta 4861 aataatctct ggtatgaatt tgatgatcag agtgtcactg aagtttcaga atctactgta 4921 caaaatgcag aagcttacgt tcttttctat aggaagagca gcgaagaggc acaaaaagag 4981 aggagaagga tatcaaattt attgaacata atggaaccaa gcctccttca gttttatatt 5041 tctcgacagt ggcttaataa atttaagacc tttgccgaac ctggccctat ttcaaataat 5101 gactttcttt gtattcatgg aggtgttcct ccaagaaaag ctggttatat tgaagacctg 5161 gttttgatgc tgcctcagaa catttgggat aacctatata gcaggtatgg tggaggacca 5221 gctgtcaacc atctgtacat ttgtcatact tgccaaattg aggcggagaa aattgaaaaa 5281 agaagaaaaa ctgaattgga aatttttatt cggcttaaca gagcgttcca aaaagaggac 5341 tctccagcta ctttttattg catcagtatg cagtggttta gagaatggga aagttttgtg 5401 aagggtaaag atggagatcc tccaggtcct attgacaata ctaagattgc agtcactaaa 5461 tgtggtaatg tgatgcttag gcaaggagca gattctggcc agatttctga agaaacatgg 5521 aattttctgc agtctattta tggtggaggg cctgaagtta tcctgcgacc tccggttgtt 5581 catgttgatc cagatatact tcaagcagaa gaaaaaattg aagtagaaac tcggtctttg 5641 ctcgaggact acaaagacca tgacggtgat tataaagatc atgacatcga ctacaaggat 5701 gacgatgaca agtagcggcc gcggatcccg cccctctccc tccccccccc ctaacgttac 5761 tggccgaagc cgcttggaat aaggccggtg tgcgtttgtc tatatgttat tttccaccat 5821 attgccgtct tttggcaatg tgagggcccg gaaacctggc cctgtcttct tgacgagcat 5881 tcctaggggt ctttcccctc tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga 5941 agcagttcct ctggaagctt cttgaagaca aacaacgtct gtagcgaccc tttgcaggca 6001 gcggaacccc ccacctggcg acaggtgcct ctgcggccaa aagccacgtg tataagatac 6061 acctgcaaag gcggcacaac cccagtgcca cgttgtgagt tggatagttg tggaaagagt 6121 caaatggctc tcctcaagcg tattcaacaa ggggctgaag gatgcccaga aggtacccca 6181 ttgtatggga tctgatctgg ggcctcggtg cacatgcttt acatgtgttt agtcgaggtt 6241 aaaaaaacgt ctaggccccc cgaaccacgg ggacgtggtt ttcctttgaa aaacacgatg 6301 ataagcttgc cacaacccac aaggagacga ccttccatga ccgagtacaa gcccacggtg 6361 cgcctcgcca cccgcgacga cgtcccccgg gccgtacgca ccctcgccgc cgcgttcgcc 6421 gactaccccg ccacgcgcca caccgtcgac ccggaccgcc acatcgagcg ggtcaccgag 6481 ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac 6541 gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc 6601 gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc gcagcaacag 6661 atggaaggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct ggccaccgtc 6721 ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg 6781 gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc 6841 cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg 6901 cgcacctggt gcatgacccg caagcccggt gcctagacgc gtctggaaca atcaacctct 6961 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct 7021 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat 7081 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt 7141 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat 7201 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc 7261 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga 7321 caattccgtg gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc 7381 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga 7441 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc 7501 tcagacgagt cggatctccc tttgggccgc ctccccgcct ggaattaatt ctgcagtcga 7561 gacctagaaa aacatggagc aatcacaagt agcaatacag cagctaccaa tgctgattgt 7621 gcctggctag aagcacaaga ggaggaggag gtgggttttc cagtcacacc tcaggtacct 7681 ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagagg 7741 ggactggaag ggctaattca ctcccaacga agacaagata tccttgatct gtggatctac 7801 cacacacaag gctacttccc tgattagcag aactacacac cagggccagg ggtcagatat 7861 ccactgacct ttggatggtg ctacaagcta gtaccagttg agccagataa ggtagaagag 7921 gccaataaag gagagaacac cagcttgtta caccctgtga gcctgcatgg gatggatgac 7981 ccggagagag aagtgttaga gtggaggttt gacagccgcc tagcatttca tcacgtggcc 8041 cgagagctgc atccggagta cttcaagaac tgctgatatc gagcttgcta caagggactt 8101 tccgctgggg actttccagg gaggcgtggc ctgggcggga ctggggagtg gcgagccctc 8161 agatcctgca tataagcagc tgctttttgc ctgtactggg tctctctggt tagaccagat 8221 ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt 8281 gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc 8341 cctcagaccc ttttagtcag tgtggaaaat ctctagcagt agtagttcat gtcatcttat 8401 tattcagtat ttataacttg caaagaaatg aatatcagag agtgagaggc cttgacattg 8461 ctagcgttta ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc 8521 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg 8581 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc 8641 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg 8701 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc 8761 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac 8821 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa 8881 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca 8941 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc 9001 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata 9061 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta 9121 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca 9181 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga 9241 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg 9301 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg 9361 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg 9421 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag 9481 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa 9541 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat 9601 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc 9661 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc 9721 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc 9781 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc 9841 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc 9901 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt 9961 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc 10021 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa 10081 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt 10141 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg 10201 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc 10261 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa 10321 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt 10381 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt 10441 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag 10501 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta 10561 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat 10621 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatca 10681 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa 10741 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt 10801 atcatgtctg gatcaactgg ataactcaag ctaaccaaaa tcatcccaaa cttcccaccc 10861 cataccctat taccactgcc aattacctgt ggtttcattt actctaaacc tgtgattcct 10921 ctgaattatt ttcattttaa agaaattgta tttgttaaat atgtactaca aacttagtag 10981 tttttaaaga aattgtattt gttaaatatg tactacaaac ttagtagt //