LOCUS V021552 9068 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V021552 VERSION V021552 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 2826..3794 /label="CD47(NM_001777)" /note="CD47(NM_001777)" /gene="CD47" CDS 3801..3824 /codon_start=1 /product="FLAG® epitope tag, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDDDDK" /label="FLAG" promoter 3856..4355 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 4376..4975 /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 4989..5577 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5784..6417 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(6546..6562) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6570..6586 /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(6594..6624) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6639..6660 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6948..7536) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7707..8567) /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(8568..8672) /gene="bla" /label="AmpR promoter" polyA_signal 8720..8854 /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 ccaccatgtg gcccctggta gcggcgctgt tgctgggctc ggcgtgctgc ggatcagctc 2881 agctactatt taataaaaca aaatctgtag aattcacgtt ttgtaatgac actgtcgtca 2941 ttccatgctt tgttactaat atggaggcac aaaacactac tgaagtatac gtaaagtgga 3001 aatttaaagg aagagatatt tacacctttg atggagctct aaacaagtcc actgtcccca 3061 ctgactttag tagtgcaaaa attgaagtct cacaattact aaaaggagat gcctctttga 3121 agatggataa gagtgatgct gtctcacaca caggaaacta cacttgtgaa gtaacagaat 3181 taaccagaga aggtgaaacg atcatcgagc taaaatatcg tgttgtttca tggttttctc 3241 caaatgaaaa tattcttatt gttattttcc caatttttgc tatactcctg ttctggggac 3301 agtttggtat taaaacactt aaatatagat ccggtggtat ggatgagaaa acaattgctt 3361 tacttgttgc tggactagtg atcactgtca ttgtcattgt tggagccatt cttttcgtcc 3421 caggtgaata ttcattaaag aatgctactg gccttggttt aattgtgact tctacaggga 3481 tattaatatt acttcactac tatgtgttta gtacagcgat tggattaacc tccttcgtca 3541 ttgccatatt ggttattcag gtgatagcct atatcctcgc tgtggttgga ctgagtctct 3601 gtattgcggc gtgtatacca atgcatggcc ctcttctgat ttcaggtttg agtatcttag 3661 ctctagcaca attacttgga ctagtttata tgaaatttgt ggcttccaat cagaagacta 3721 tacaacctcc taggaaagct gtagaggaac cccttaatgc attcaaagaa tcaaaaggaa 3781 tgatgaatga tgaagaattc gattacaagg atgacgacga taagtgagga tcccgcgact 3841 ctagataatt ctaccgggta ggggaggcgc ttttcccaag gcagtctgga gcatgcgctt 3901 tagcagcccc gctgggcact tggcgctaca caagtggcct ctggcctcgc acacattcca 3961 catccaccgg taggcgccaa ccggctccgt tctttggtgg ccccttcgcg ccaccttcta 4021 ctcctcccct agtcaggaag ttcccccccg ccccgcagct cgcgtcgtgc aggacgtgac 4081 aaatggaagt agcacgtctc actagtctcg tgcagatgga cagcaccgct gagcaatgga 4141 agcgggtagg cctttggggc agcggccaat agcagctttg ctccttcgct ttctgggctc 4201 agaggctggg aaggggtggg tccgggggcg ggctcagggg cgggctcagg ggcggggcgg 4261 gcgcccgaag gtcctccgga ggcccggcat tctgcacgct tcaaaagcgc acgtctgccg 4321 cgctgttctc ctcttcctca tctccgggcc tttcgacctg cagcccaagc ttaccatgac 4381 cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac 4441 cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca 4501 catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg 4561 caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt 4621 cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg 4681 gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc 4741 gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc 4801 cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac 4861 ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt 4921 cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgaccgcg 4981 tctggaacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact 5041 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg 5101 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg 5161 aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa 5221 cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc 5281 ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg 5341 ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaagctgacg tcctttccat 5401 ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt 5461 cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc 5521 cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg 5581 gaattaattc tgcagtcgag acctagaaaa acatggagca atcacaagta gcaatacagc 5641 agctaccaat gctgattgtg cctggctaga agcacaagag gaggaggagg tgggttttcc 5701 agtcacacct caggtacctt taagaccaat gacttacaag gcagctgtag atcttagcca 5761 ctttttaaaa gaaaagaggg gactggaagg gctaattcac tcccaacgaa gacaagatat 5821 ccttgatctg tggatctacc acacacaagg ctacttccct gattagcaga actacacacc 5881 agggccaggg gtcagatatc cactgacctt tggatggtgc tacaagctag taccagttga 5941 gccagataag gtagaagagg ccaataaagg agagaacacc agcttgttac accctgtgag 6001 cctgcatggg atggatgacc cggagagaga agtgttagag tggaggtttg acagccgcct 6061 agcatttcat cacgtggccc gagagctgca tccggagtac ttcaagaact gctgatatcg 6121 agcttgctac aagggacttt ccgctgggga ctttccaggg aggcgtggcc tgggcgggac 6181 tggggagtgg cgagccctca gatcctgcat ataagcagct gctttttgcc tgtactgggt 6241 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc 6301 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg 6361 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta 6421 gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga 6481 gtgagaggcc ttgacattgc tagcgtttta ccgtcgacct ctagctagag cttggcgtaa 6541 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata 6601 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta 6661 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa 6721 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg 6781 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag 6841 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa 6901 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc 6961 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca 7021 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg 7081 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct 7141 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt 7201 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag 7261 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc 7321 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac 7381 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga 7441 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc 7501 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg 7561 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca 7621 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt 7681 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca 7741 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg 7801 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca 7861 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt 7921 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt 7981 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca 8041 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca 8101 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga 8161 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact 8221 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga 8281 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg 8341 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc 8401 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga 8461 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat 8521 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt 8581 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt 8641 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac 8701 gtcgacggat cgggagatca acttgtttat tgcagcttat aatggttaca aataaagcaa 8761 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc 8821 caaactcatc aatgtatctt atcatgtctg gatcaactgg ataactcaag ctaaccaaaa 8881 tcatcccaaa cttcccaccc cataccctat taccactgcc aattacctgt ggtttcattt 8941 actctaaacc tgtgattcct ctgaattatt ttcattttaa agaaattgta tttgttaaat 9001 atgtactaca aacttagtag tttttaaaga aattgtattt gttaaatatg tactacaaac 9061 ttagtagt //