LOCUS V020249 10090 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V020249 VERSION V020249 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 2820..4757 /label="WEE1(NM_003390)" /note="WEE1(NM_003390)" /gene="WEE1" CDS 4758..4781 /codon_start=1 /product="FLAG® epitope tag, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDDDDK" /label="FLAG" misc_feature 4793..5379 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" /label="IRES2" CDS 5399..5998 /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 6012..6600 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6807..7440 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(7568..7584) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7592..7608 /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(7616..7646) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7661..7682 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7970..8558) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8729..9589) /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(9590..9694) /gene="bla" /label="AmpR promoter" polyA_signal 9742..9876 /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 gtgaattcct cgagccacca 2821 tgagcttcct gagccgacag cagccgccgc caccccgccg cgccggggcg gcctgcacct 2881 tgcggcagaa gctgatcttc tcgccctgca gcgactgtga ggaggaggaa gaagaggagg 2941 aggaggaggg cagcggccac agcaccgggg aggactcggc ctttcaagag cccgactcgc 3001 cgctgccgcc cgcgcggagc cccacggagc ccgggcccga gcgccgccgc tcgcccgggc 3061 cggcccccgg cagccccggc gagctggagg aggacctgtt gctgcccggc gcctgcccgg 3121 gcgcggacga ggcgggcggt ggggcggagg gcgactcgtg ggaggaggag ggcttcggct 3181 cctcgtcgcc ggtcaagtcg ccggcggccc cctacttcct gggtagctct ttctcgccgg 3241 tgcgctgcgg cggcccagga gatgcgtcgc cgcggggttg cggggcgcgc cgggcgggcg 3301 aaggccgccg ctcgccgcgg ccggaccacc cgggcacccc gccacacaag accttccgca 3361 agctgcgact cttcgacacc ccgcacacgc ccaagagttt gctctccaaa gctcggggaa 3421 ttgattccag ctctgttaaa ctccggggta gttctctctt catggataca gaaaaatcag 3481 gaaaaaggga atttgatgtg cgacagactc ctcaagtgaa tattaatcct tttactccgg 3541 attctttgtt gcttcattcc tcaggacagt gtcgtcgtag aaagagaacg tattggaatg 3601 attcctgtgg tgaagacatg gaagccagtg attatgagct tgaagatgaa acaagacctg 3661 ctaagagaat tacaattact gaaagcaata tgaagtcccg gtatacaaca gaatttcatg 3721 agctagagaa aatcggctct ggagaatttg gttctgtatt taagtgtgtg aagaggctgg 3781 atggatgcat ttatgccatt aagcgatcaa aaaagccatt ggcgggctct gttgatgagc 3841 agaacgcttt gagagaagta tatgctcatg cagtgcttgg acagcattct catgtagttc 3901 gatatttctc tgcgtgggca gaagatgatc atatgcttat acagaatgaa tattgtaatg 3961 gtggaagttt agctgatgct ataagtgaaa actacagaat catgagttac tttaaagaag 4021 cagagttgaa ggatctcctt ttgcaagttg gccgaggctt gaggtatatt cattcaatgt 4081 ctttggttca catggatata aaacctagta atattttcat atctcgaacc tcaatcccaa 4141 atgctgcctc tgaagaagga gacgaagatg attgggcatc caacaaagtt atgtttaaaa 4201 taggtgatct tgggcatgta acaaggatct ccagtccaca agttgaagag ggcgatagtc 4261 gttttcttgc aaatgaagtt ttacaggaga attataccca tctaccaaaa gcagatattt 4321 ttgcgcttgc cctcacagtg gtatgtgctg ctggtgctga acctcttccg agaaatggag 4381 atcaatggca tgaaatcaga cagggtagat tacctcggat accacaagtg ctttcccaag 4441 aatttacaga gttgctaaaa gttatgattc atccagatcc agagagaaga ccttcagcaa 4501 tggcactggt aaagcattca gtattgctgt ccgcttctag aaagagtgca gaacaattac 4561 gaatagaatt gaatgccgaa aagttcaaaa attcactttt acaaaaagaa ctcaagaaag 4621 cacagatggc aaaagctgca gctgaggaaa gagcactctt cactgaccgg atggccacta 4681 ggtccaccac ccagagtaat agaacatctc gacttattgg aaagaaaatg aaccgctctg 4741 tcagccttac tatatacgat tacaaggacg acgatgacaa gtgaggatcc cgcccctctc 4801 cctccccccc ccctaacgtt actggccgaa gccgcttgga ataaggccgg tgtgcgtttg 4861 tctatatgtt attttccacc atattgccgt cttttggcaa tgtgagggcc cggaaacctg 4921 gccctgtctt cttgacgagc attcctaggg gtctttcccc tctcgccaaa ggaatgcaag 4981 gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagc ttcttgaaga caaacaacgt 5041 ctgtagcgac cctttgcagg cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc 5101 aaaagccacg tgtataagat acacctgcaa aggcggcaca accccagtgc cacgttgtga 5161 gttggatagt tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac aaggggctga 5221 aggatgccca gaaggtaccc cattgtatgg gatctgatct ggggcctcgg tgcacatgct 5281 ttacatgtgt ttagtcgagg ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg 5341 ttttcctttg aaaaacacga tgataagctt gccacaaccc acaaggagac gaccttccat 5401 gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtccccc gggccgtacg 5461 caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg acccggaccg 5521 ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat 5581 cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag 5641 cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc 5701 ccggctggcc gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc 5761 cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag 5821 cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga 5881 gacctccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga 5941 cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctagac 6001 gcgtctggaa caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta 6061 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta 6121 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt 6181 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg 6241 caacccccac tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt 6301 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag 6361 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc 6421 catggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc 6481 cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc 6541 ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc 6601 ctggaattaa ttctgcagtc gagacctaga aaaacatgga gcaatcacaa gtagcaatac 6661 agcagctacc aatgctgatt gtgcctggct agaagcacaa gaggaggagg aggtgggttt 6721 tccagtcaca cctcaggtac ctttaagacc aatgacttac aaggcagctg tagatcttag 6781 ccacttttta aaagaaaaga ggggactgga agggctaatt cactcccaac gaagacaaga 6841 tatccttgat ctgtggatct accacacaca aggctacttc cctgattagc agaactacac 6901 accagggcca ggggtcagat atccactgac ctttggatgg tgctacaagc tagtaccagt 6961 tgagccagat aaggtagaag aggccaataa aggagagaac accagcttgt tacaccctgt 7021 gagcctgcat gggatggatg acccggagag agaagtgtta gagtggaggt ttgacagccg 7081 cctagcattt catcacgtgg cccgagagct gcatccggag tacttcaaga actgctgata 7141 tcgagcttgc tacaagggac tttccgctgg ggactttcca gggaggcgtg gcctgggcgg 7201 gactggggag tggcgagccc tcagatcctg catataagca gctgcttttt gcctgtactg 7261 ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac 7321 tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt 7381 gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca 7441 gtagtagttc atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag 7501 agagtgagag gccttgacat tgctagcgtt taccgtcgac ctctagctag agcttggcgt 7561 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca 7621 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat 7681 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt 7741 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct 7801 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa 7861 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa 7921 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc 7981 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga 8041 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc 8101 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt 8161 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct 8221 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg 8281 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta 8341 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct 8401 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa 8461 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt 8521 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta 8581 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat 8641 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa 8701 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct 8761 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta 8821 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct 8881 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg 8941 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa 9001 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt 9061 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta 9121 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca 9181 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta 9241 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct 9301 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg 9361 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac 9421 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact 9481 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa 9541 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt 9601 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat 9661 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg 9721 acgtcgacgg atcgggagat caacttgttt attgcagctt ataatggtta caaataaagc 9781 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg 9841 tccaaactca tcaatgtatc ttatcatgtc tggatcaact ggataactca agctaaccaa 9901 aatcatccca aacttcccac cccataccct attaccactg ccaattacct gtggtttcat 9961 ttactctaaa cctgtgattc ctctgaatta ttttcatttt aaagaaattg tatttgttaa 10021 atatgtacta caaacttagt agtttttaaa gaaattgtat ttgttaaata tgtactacaa 10081 acttagtagt //