LOCUS V022497 9903 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V022497 VERSION V022497 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..4374 /label="USP3(NM_006537)" /note="USP3(NM_006537)" /gene="USP3" CDS 4381..4446 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 4477..5063 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" /label="IRES2" CDS 5064..5858 /codon_start=1 /gene="aph(3')-II (or nptII)" /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin®)" /product="aminoglycoside phosphotransferase from Tn5" /transl_table=1 /translation="MAEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*" /label="NeoR/KanR" misc_feature 5872..6460 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6667..7300 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(7428..7444) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7452..7468 /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(7476..7506) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7521..7542 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7830..8418) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8589..9449) /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(9450..9554) /gene="bla" /label="AmpR promoter" polyA_signal 9602..9736 /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 caccatggag 2821 tgtccacacc tgagctccag cgtctgcatt gctccggact cagccaagtt ccccaacggc 2881 tccccgtcgt cctggtgctg cagcgtgtgc cggtccaaca aaagcccttg ggtctgtttg 2941 acttgttcaa gtgtccactg tggaaggtat gtgaatggcc atgcaaaaaa acattatgaa 3001 gatgcacaag tacctttaac caaccataag aaatcagaaa agcaagataa agttcagcac 3061 acagtatgta tggattgcag tagctacagt acatactgtt atcgctgtga tgattttgtg 3121 gttaatgaca ccaagctggg actggtacag aaagtcagag aacacttaca gaacttggaa 3181 aactcagctt tcacagctga caggcataag aaaagaaaac ttttggaaaa ctcaacacta 3241 aacagcaagt tattaaaagt aaatggaagc accactgcca tttgtgccac aggccttcgg 3301 aatttgggga acacatgttt catgaatgcc atccttcagt cactcagtaa cattgagcag 3361 ttttgctgtt atttcaaaga actgcccgcc gtggagttaa ggaatgggaa aacagcagga 3421 aggcggacat accacaccag gagccaaggg gataacaatg tgtctttggt agaagagttt 3481 agaaagacac tctgtgcttt atggcaaggc agccagactg catttagccc agagtcctta 3541 ttttatgttg tttggaagat tatgccaaac tttaggggct atcaacagca ggacgcccat 3601 gaattcatgc gctacctttt ggaccaccta cacttggaac ttcagggcgg tttcaacggt 3661 gtttcccgct cagcaattct gcaggagaat tctactctgt ctgcaagtaa caagtgttgc 3721 ataaatggag catctactgt tgtcacggct atattcggag gcattctcca aaatgaggtt 3781 aactgcctca tatgtgggac agaatctaga aagtttgatc cattcctaga cctttcatta 3841 gatattccaa gtcagttcag aagtaagcgc tctaagaatc aagaaaatgg accagtttgt 3901 tcgttacgag attgtcttcg cagttttacc gacttagaag aacttgatga gacagagtta 3961 tatatgtgcc ataaatgcaa aaagaaacaa aagtccacaa aaaagttttg gattcaaaaa 4021 ctacccaagg tgctatgctt acatttgaaa agatttcatt ggacagcata tttaagaaat 4081 aaagttgata catacgtaga atttccactg agaggcctag acatgaaatg ctacttacta 4141 gagcctgaga acagtggccc ggagagctgc ctgtatgacc tcgccgctgt ggtggtgcac 4201 catggttccg gggttggttc tggacattac acagcatacg caactcacga aggccgctgg 4261 ttccacttca atgacagtac tgtaacactg actgacgagg agactgtggt gaaggcgaag 4321 gcctacatcc ttttctacgt ggaacaccag gccaaagctg gatcggataa acttctcgag 4381 gactacaaag accatgacgg tgattataaa gatcatgaca tcgactacaa ggatgacgat 4441 gacaagtaga ctagttctag agcggccgcg gatccgcccc tctccctccc ccccccctaa 4501 cgttactggc cgaagccgct tggaataagg ccggtgtgcg tttgtctata tgttattttc 4561 caccatattg ccgtcttttg gcaatgtgag ggcccggaaa cctggccctg tcttcttgac 4621 gagcattcct aggggtcttt cccctctcgc caaaggaatg caaggtctgt tgaatgtcgt 4681 gaaggaagca gttcctctgg aagcttcttg aagacaaaca acgtctgtag cgaccctttg 4741 caggcagcgg aaccccccac ctggcgacag gtgcctctgc ggccaaaagc cacgtgtata 4801 agatacacct gcaaaggcgg cacaacccca gtgccacgtt gtgagttgga tagttgtgga 4861 aagagtcaaa tggctctcct caagcgtatt caacaagggg ctgaaggatg cccagaaggt 4921 accccattgt atgggatctg atctggggcc tcggtgcaca tgctttacat gtgtttagtc 4981 gaggttaaaa aaacgtctag gccccccgaa ccacggggac gtggttttcc tttgaaaaac 5041 acgatgataa gcttgccaca accatggctg aacaagatgg attgcacgca ggttctccgg 5101 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg 5161 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc 5221 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga 5281 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc 5341 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag 5401 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat 5461 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg 5521 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca 5581 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct 5641 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg 5701 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg 5761 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc 5821 gcatcgcctt ctatcgcctt cttgacgagt tcttctgaac gcgtctggaa caatcaacct 5881 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg 5941 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc 6001 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt 6061 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc 6121 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg 6181 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact 6241 gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct cgcctgtgtt 6301 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg 6361 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc 6421 cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggaattaa ttctgcagtc 6481 gagacctaga aaaacatgga gcaatcacaa gtagcaatac agcagctacc aatgctgatt 6541 gtgcctggct agaagcacaa gaggaggagg aggtgggttt tccagtcaca cctcaggtac 6601 ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaaga 6661 ggggactgga agggctaatt cactcccaac gaagacaaga tatccttgat ctgtggatct 6721 accacacaca aggctacttc cctgattagc agaactacac accagggcca ggggtcagat 6781 atccactgac ctttggatgg tgctacaagc tagtaccagt tgagccagat aaggtagaag 6841 aggccaataa aggagagaac accagcttgt tacaccctgt gagcctgcat gggatggatg 6901 acccggagag agaagtgtta gagtggaggt ttgacagccg cctagcattt catcacgtgg 6961 cccgagagct gcatccggag tacttcaaga actgctgata tcgagcttgc tacaagggac 7021 tttccgctgg ggactttcca gggaggcgtg gcctgggcgg gactggggag tggcgagccc 7081 tcagatcctg catataagca gctgcttttt gcctgtactg ggtctctctg gttagaccag 7141 atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc 7201 ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga 7261 tccctcagac ccttttagtc agtgtggaaa atctctagca gtagtagttc atgtcatctt 7321 attattcagt atttataact tgcaaagaaa tgaatatcag agagtgagag gccttgacat 7381 tgctagcgtt taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt 7441 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag 7501 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg 7561 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg 7621 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc 7681 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc 7741 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg 7801 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat 7861 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag 7921 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga 7981 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg 8041 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt 8101 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac 8161 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc 8221 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt 8281 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc 8341 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc 8401 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg 8461 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag 8521 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg 8581 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt 8641 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca 8701 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca 8761 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc 8821 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt 8881 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg 8941 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc 9001 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg 9061 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga 9121 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga 9181 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta 9241 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg 9301 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact 9361 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata 9421 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt 9481 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa 9541 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat 9601 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac 9661 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc 9721 ttatcatgtc tggatcaact ggataactca agctaaccaa aatcatccca aacttcccac 9781 cccataccct attaccactg ccaattacct gtggtttcat ttactctaaa cctgtgattc 9841 ctctgaatta ttttcatttt aaagaaattg tatttgttaa atatgtacta caaacttagt 9901 agt //