LOCUS V022361 8672 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V022361 VERSION V022361 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..3873 /label="WNT7B(NM_058238)" /note="WNT7B(NM_058238)" /gene="WNT7B" CDS 3895..3918 /codon_start=1 /product="FLAG® epitope tag, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDDDDK" /label="FLAG" CDS 3928..3981 /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 4012..4719 /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 4726..5314 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5388..6021 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(6150..6166) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6174..6190 /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(6198..6228) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6243..6264 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6552..7140) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7311..8171) /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(8172..8276) /gene="bla" /label="AmpR promoter" polyA_signal 8324..8458 /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 acagaaactt tcgcaagtgg attttctacg tgtttctctg ctttggcgtc 2881 ctgtacgtga agctcggagc actgtcatcc gtggtggccc tgggagccaa catcatctgc 2941 aacaagattc ctggcctagc cccgcggcag cgtgccatct gccagagtcg gcccgatgcc 3001 atcattgtga ttggggaggg ggcgcagatg ggcatcaacg agtgccagta ccagttccgc 3061 ttcggacgct ggaactgctc tgccctcggc gagaagaccg tcttcgggca agagctccga 3121 gtagggagcc gtgaggctgc cttcacgtac gccatcaccg cggctggcgt ggcgcacgcc 3181 gtcaccgctg cctgcagcca agggaacctg agcaactgcg gctgcgaccg cgagaagcag 3241 ggctactaca accaagccga gggctggaag tggggcggct gctcggccga cgtgcgttac 3301 ggcatcgact tctcccggcg cttcgtggac gctcgggaga tcaagaagaa cgcgcggcgc 3361 ctcatgaacc tgcataacaa tgaggccggc aggaaggttc tagaggaccg gatgcagctg 3421 gagtgcaagt gccacggcgt gtctggctcc tgcaccacca aaacctgctg gaccacgctg 3481 cccaagttcc gagaggtggg ccacctgctg aaggagaagt acaacgcggc cgtgcaggtg 3541 gaggtggtgc gggccagccg tctgcggcag cccaccttcc tgcgcatcaa acagctgcgc 3601 agctatcaga agcccatgga gacagacctg gtgtacattg agaagtcgcc caactactgc 3661 gaggaggacg cggccacggg cagcgtgggc acgcagggcc gtctctgcaa ccgcacgtcg 3721 cccggcgcgg acggctgtga caccatgtgc tgcggccgag gctacaacac ccaccagtac 3781 accaaggtgt ggcagtgcaa ctgcaaattc cactggtgct gcttcgtcaa gtgcaacacc 3841 tgcagcgagc gcaccgaggt cttcacctgc aaggaattcg gaggaggggg aagcgactac 3901 aaggacgacg atgacaaggg cagcggtgag ggcagaggaa gtcttctaac atgcggtgac 3961 gtggaggaga atcccggccc ttccggaatg gagagcgacg agagcggcct ggtgagcaag 4021 ggcgaggagg ataacatggc catcatcaag gagttcatgc gcttcaaggt gcacatggag 4081 ggctccgtga acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag 4141 ggcacccaga ccgccaagct gaaggtgacc aagggtggcc ccctgccctt cgcctgggac 4201 atcctgtccc ctcagttcat gtacggctcc aaggcctacg tgaagcaccc cgccgacatc 4261 cccgactact tgaagctgtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc 4321 gaggacggcg gcgtggtgac cgtgacccag gactcctccc tgcaggacgg cgagttcatc 4381 tacaaggtga agctgcgcgg caccaacttc ccctccgacg gccccgtaat gcagaagaag 4441 accatgggct gggaggcctc ctccgagcgg atgtaccccg aggacggcgc cctgaagggc 4501 gagatcaagc agaggctgaa gctgaaggac ggcggccact acgacgctga ggtcaagacc 4561 acctacaagg ccaagaagcc cgtgcagctg cccggcgcct acaacgtcaa catcaagttg 4621 gacatcacct cccacaacga ggactacacc atcgtggaac agtacgaacg cgccgagggc 4681 cgccactcca ccggcggcat ggacgagctg tacaagtagg tcgacaatca acctctggat 4741 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt 4801 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc 4861 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg 4921 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc 4981 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa 5041 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat 5101 tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc 5161 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt 5221 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag 5281 acgagtcgga tctccctttg ggccgcctcc ccgcctggta cctttaagac caatgactta 5341 caaggcagct gtagatctta gccacttttt aaaagaaaag aggggactgg aagggctaat 5401 tcactcccaa cgaagacaag atatccttga tctgtggatc taccacacac aaggctactt 5461 ccctgattag cagaactaca caccagggcc aggggtcaga tatccactga cctttggatg 5521 gtgctacaag ctagtaccag ttgagccaga taaggtagaa gaggccaata aaggagagaa 5581 caccagcttg ttacaccctg tgagcctgca tgggatggat gacccggaga gagaagtgtt 5641 agagtggagg tttgacagcc gcctagcatt tcatcacgtg gcccgagagc tgcatccgga 5701 gtacttcaag aactgctgat atcgagcttg ctacaaggga ctttccgctg gggactttcc 5761 agggaggcgt ggcctgggcg ggactgggga gtggcgagcc ctcagatcct gcatataagc 5821 agctgctttt tgcctgtact gggtctctct ggttagacca gatctgagcc tgggagctct 5881 ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag 5941 tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt 6001 cagtgtggaa aatctctagc agtagtagtt catgtcatct tattattcag tatttataac 6061 ttgcaaagaa atgaatatca gagagtgaga ggccttgaca ttgctagcgt tttaccgtcg 6121 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat 6181 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc 6241 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga 6301 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt 6361 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg 6421 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac 6481 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg 6541 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca 6601 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc 6661 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc 6721 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag 6781 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc 6841 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca 6901 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg 6961 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg 7021 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct 7081 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa 7141 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa 7201 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa 7261 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc 7321 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga 7381 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca 7441 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc 7501 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat 7561 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc 7621 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt 7681 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc 7741 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg 7801 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt 7861 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg 7921 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga 7981 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg 8041 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg 8101 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt 8161 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc 8221 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca 8281 tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atcaacttgt ttattgcagc 8341 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc 8401 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatcaa 8461 ctggataact caagctaacc aaaatcatcc caaacttccc accccatacc ctattaccac 8521 tgccaattac ctgtggtttc atttactcta aacctgtgat tcctctgaat tattttcatt 8581 ttaaagaaat tgtatttgtt aaatatgtac tacaaactta gtagttttta aagaaattgt 8641 atttgttaaa tatgtactac aaacttagta gt //