LOCUS V022363 8798 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V022363 VERSION V022363 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 2827..3999 /label="WNT2B(NM_024494)" /note="WNT2B(NM_024494)" /gene="WNT2B" CDS 4021..4044 /codon_start=1 /product="FLAG® epitope tag, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDDDDK" /label="FLAG" CDS 4054..4107 /codon_start=1 /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /product="2A peptide from Thosea asigna virus capsid protein" /protein_id="" /transl_table=1 /translation="EGRGSLLTCGDVEENPGP" /label="T2A" CDS 4138..4845 /codon_start=1 /note="mammalian codon-optimized" /product="monomeric derivative of DsRed fluorescent protein (Shaner et al., 2004)" /transl_table=1 /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE GRHSTGGMDELYK*" /label="mCherry" misc_feature 4852..5440 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5514..6147 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(6276..6292) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6300..6316 /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(6324..6354) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6369..6390 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6678..7266) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7437..8297) /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(8298..8402) /gene="bla" /label="AmpR promoter" polyA_signal 8450..8584 /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 gccaccatgc tgagaccggg tggtgcggag gaagctgcgc agctcccgct tcggcgcgcc 2881 agcgccccgg tccctgtgcc gtcgcccgcg gcccccgacg gctcccgggc ttcggcccgc 2941 ctaggtcttg cctgccttct gctcctgctg ctgctgacgc tgccggcccg cgtagacacg 3001 tcctggtggt acattggggc actgggggca cgagtgatct gtgacaatat ccctggtttg 3061 gtgagccggc agcggcagct gtgccagcgt tacccagaca tcatgcgttc agtgggcgag 3121 ggtgcccgag aatggatccg agagtgtcag caccaattcc gccaccaccg ctggaactgt 3181 accaccctgg accgggacca caccgtcttt ggccgtgtca tgctcagaag tagccgagag 3241 gcagcttttg tatatgccat ctcatcagca ggggtagtcc acgctattac tcgcgcctgt 3301 agccagggtg aactgagtgt gtgcagctgt gacccctaca cccgtggccg acaccatgac 3361 cagcgtgggg actttgactg gggtggctgc agtgacaaca tccactacgg tgtccgtttt 3421 gccaaggcct tcgtggatgc caaggagaag aggcttaagg atgcccgggc cctcatgaac 3481 ttacataata accgctgtgg tcgcacggct gtgcggcggt ttctgaagct ggagtgtaag 3541 tgccatggcg tgagtggttc ctgtactctg cgcacctgct ggcgtgcact ctcagatttc 3601 cgccgcacag gtgattacct gcggcgacgc tatgatgggg ctgtgcaggt gatggccacc 3661 caagatggtg ccaacttcac cgcagcccgc caaggctatc gccgtgccac ccggactgat 3721 cttgtctact ttgacaactc tccagattac tgtgtcttgg acaaggctgc aggttcccta 3781 ggcactgcag gccgtgtctg cagcaagaca tcaaaaggaa cagacggttg tgaaatcatg 3841 tgctgtggcc gagggtacga cacaactcga gtcacccgtg ttacccagtg tgagtgcaaa 3901 ttccactggt gctgtgctgt acggtgcaag gaatgcagaa atactgtgga cgtccatact 3961 tgcaaagccc ccaagaaggc agagtggctg gaccaaaccg aattcggagg agggggaagc 4021 gactacaagg acgacgatga caagggcagc ggtgagggca gaggaagtct tctaacatgc 4081 ggtgacgtgg aggagaatcc cggcccttcc ggaatggaga gcgacgagag cggcctggtg 4141 agcaagggcg aggaggataa catggccatc atcaaggagt tcatgcgctt caaggtgcac 4201 atggagggct ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc 4261 tacgagggca cccagaccgc caagctgaag gtgaccaagg gtggccccct gcccttcgcc 4321 tgggacatcc tgtcccctca gttcatgtac ggctccaagg cctacgtgaa gcaccccgcc 4381 gacatccccg actacttgaa gctgtccttc cccgagggct tcaagtggga gcgcgtgatg 4441 aacttcgagg acggcggcgt ggtgaccgtg acccaggact cctccctgca ggacggcgag 4501 ttcatctaca aggtgaagct gcgcggcacc aacttcccct ccgacggccc cgtaatgcag 4561 aagaagacca tgggctggga ggcctcctcc gagcggatgt accccgagga cggcgccctg 4621 aagggcgaga tcaagcagag gctgaagctg aaggacggcg gccactacga cgctgaggtc 4681 aagaccacct acaaggccaa gaagcccgtg cagctgcccg gcgcctacaa cgtcaacatc 4741 aagttggaca tcacctccca caacgaggac tacaccatcg tggaacagta cgaacgcgcc 4801 gagggccgcc actccaccgg cggcatggac gagctgtaca agtaggtcga caatcaacct 4861 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg 4921 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc 4981 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt 5041 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc 5101 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg 5161 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact 5221 gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt 5281 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg 5341 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc 5401 cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggtacctt taagaccaat 5461 gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaagaggg gactggaagg 5521 gctaattcac tcccaacgaa gacaagatat ccttgatctg tggatctacc acacacaagg 5581 ctacttccct gattagcaga actacacacc agggccaggg gtcagatatc cactgacctt 5641 tggatggtgc tacaagctag taccagttga gccagataag gtagaagagg ccaataaagg 5701 agagaacacc agcttgttac accctgtgag cctgcatggg atggatgacc cggagagaga 5761 agtgttagag tggaggtttg acagccgcct agcatttcat cacgtggccc gagagctgca 5821 tccggagtac ttcaagaact gctgatatcg agcttgctac aagggacttt ccgctgggga 5881 ctttccaggg aggcgtggcc tgggcgggac tggggagtgg cgagccctca gatcctgcat 5941 ataagcagct gctttttgcc tgtactgggt ctctctggtt agaccagatc tgagcctggg 6001 agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc 6061 ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct 6121 tttagtcagt gtggaaaatc tctagcagta gtagttcatg tcatcttatt attcagtatt 6181 tataacttgc aaagaaatga atatcagaga gtgagaggcc ttgacattgc tagcgtttta 6241 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat 6301 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg 6361 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag 6421 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt 6481 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg 6541 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg 6601 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag 6661 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga 6721 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct 6781 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc 6841 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg 6901 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc 6961 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca 7021 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag 7081 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct 7141 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc 7201 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga 7261 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca 7321 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat 7381 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac 7441 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt 7501 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt 7561 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag 7621 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct 7681 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt 7741 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc 7801 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt 7861 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg 7921 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg 7981 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct 8041 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc 8101 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt 8161 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt 8221 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg 8281 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat 8341 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg 8401 cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatca acttgtttat 8461 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt 8521 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg 8581 gatcaactgg ataactcaag ctaaccaaaa tcatcccaaa cttcccaccc cataccctat 8641 taccactgcc aattacctgt ggtttcattt actctaaacc tgtgattcct ctgaattatt 8701 ttcattttaa agaaattgta tttgttaaat atgtactaca aacttagtag tttttaaaga 8761 aattgtattt gttaaatatg tactacaaac ttagtagt //