LOCUS V032467 8908 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V032467 VERSION V032467 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 2843..3604 /label="Pgam1(NM_023418)" /note="Pgam1(NM_023418)" /gene="Pgam1" CDS 3611..3676 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3696..4195 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 4216..4815 /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 4829..5417 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5624..6257 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(6386..6402) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6410..6426 /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(6434..6464) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6479..6500 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6788..7376) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7547..8407) /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(8408..8512) /gene="bla" /label="AmpR promoter" polyA_signal 8560..8694 /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 ctcaagcttc gaattcgcca ccatggctgc ctacaagctg gtgctgatcc ggcatggcga 2881 gagcgcctgg aacctggaga accgcttcag cggctggtac gacgccgacc tgagcccggc 2941 gggccacgag gaggcgaagc gcggcgggca ggcgttgcga gatgctggct atgaatttga 3001 catctgcttc acctctgtgc agaagagagc aatccggacc ctctggacag tcctggatgc 3061 cattgaccag atgtggttgc cagtggtcag gacttggcgc ctcaatgagc gacactatgg 3121 cggtctgaca ggtctcaaca aagcagaaac tgctgctaag catggtgagg cccaggtaaa 3181 gatctggaga cgatcttatg atgtcccacc gcctcccatg gagcctgatc accccttcta 3241 cagcaacatc agcaaggatc gcaggtacgc agaccttact gaagaccagc tcccctcctg 3301 tgagagcctg aaggacacta ttgccagagc actgcccttc tggaatgaag aaattgtccc 3361 ccagatcaaa gaggggaaaa gggtcttgat tgccgcccac ggcaacagcc ttcggggcat 3421 cgttaagcat ctggagggtc tctcagaaga ggccatcatg gagctgaacc tgcccacggg 3481 catccctatc gtctatgaac tggacaagaa cttgaagccc atcaagccca tgcagttcct 3541 gggagacgag gagaccgtgc gcaaagccat ggaagctgtg gctgctcagg gcaaggtgaa 3601 gaagggatcc gactacaaag accatgacgg tgattataaa gatcatgaca tcgactacaa 3661 ggatgacgat gacaagtagt ctagataatt ctaccgggta ggggaggcgc ttttcccaag 3721 gcagtctgga gcatgcgctt tagcagcccc gctgggcact tggcgctaca caagtggcct 3781 ctggcctcgc acacattcca catccaccgg taggcgccaa ccggctccgt tctttggtgg 3841 ccccttcgcg ccaccttcta ctcctcccct agtcaggaag ttcccccccg ccccgcagct 3901 cgcgtcgtgc aggacgtgac aaatggaagt agcacgtctc actagtctcg tgcagatgga 3961 cagcaccgct gagcaatgga agcgggtagg cctttggggc agcggccaat agcagctttg 4021 ctccttcgct ttctgggctc agaggctggg aaggggtggg tccgggggcg ggctcagggg 4081 cgggctcagg ggcggggcgg gcgcccgaag gtcctccgga ggcccggcat tctgcacgct 4141 tcaaaagcgc acgtctgccg cgctgttctc ctcttcctca tctccgggcc tttcgacctg 4201 cagcccaagc ttaccatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac 4261 gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac 4321 accgtcgatc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg 4381 cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc 4441 tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg 4501 gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct cctggcgccg 4561 caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag 4621 ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg 4681 gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc 4741 ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg catgacccgc 4801 aagcccggtg cctgaccgcg tctggaacaa tcaacctctg gattacaaaa tttgtgaaag 4861 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat 4921 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc 4981 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg 5041 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct 5101 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct 5161 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg 5221 gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac 5281 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct 5341 gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct 5401 ttgggccgcc tccccgcctg gaattaattc tgcagtcgag acctagaaaa acatggagca 5461 atcacaagta gcaatacagc agctaccaat gctgattgtg cctggctaga agcacaagag 5521 gaggaggagg tgggttttcc agtcacacct caggtacctt taagaccaat gacttacaag 5581 gcagctgtag atcttagcca ctttttaaaa gaaaagaggg gactggaagg gctaattcac 5641 tcccaacgaa gacaagatat ccttgatctg tggatctacc acacacaagg ctacttccct 5701 gattagcaga actacacacc agggccaggg gtcagatatc cactgacctt tggatggtgc 5761 tacaagctag taccagttga gccagataag gtagaagagg ccaataaagg agagaacacc 5821 agcttgttac accctgtgag cctgcatggg atggatgacc cggagagaga agtgttagag 5881 tggaggtttg acagccgcct agcatttcat cacgtggccc gagagctgca tccggagtac 5941 ttcaagaact gctgatatcg agcttgctac aagggacttt ccgctgggga ctttccaggg 6001 aggcgtggcc tgggcgggac tggggagtgg cgagccctca gatcctgcat ataagcagct 6061 gctttttgcc tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg 6121 ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt 6181 gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt 6241 gtggaaaatc tctagcagta gtagttcatg tcatcttatt attcagtatt tataacttgc 6301 aaagaaatga atatcagaga gtgagaggcc ttgacattgc tagcgtttta ccgtcgacct 6361 ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc 6421 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat 6481 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc 6541 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg 6601 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag 6661 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag 6721 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc 6781 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc 6841 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc 6901 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt 6961 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg 7021 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat 7081 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag 7141 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt 7201 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc 7261 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta 7321 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag 7381 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga 7441 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa 7501 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa 7561 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc 7621 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga 7681 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa 7741 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt 7801 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg 7861 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc 7921 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg 7981 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag 8041 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt 8101 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt 8161 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac 8221 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac 8281 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag 8341 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa 8401 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga 8461 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc 8521 cccgaaaagt gccacctgac gtcgacggat cgggagatca acttgtttat tgcagcttat 8581 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg 8641 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatcaactgg 8701 ataactcaag ctaaccaaaa tcatcccaaa cttcccaccc cataccctat taccactgcc 8761 aattacctgt ggtttcattt actctaaacc tgtgattcct ctgaattatt ttcattttaa 8821 agaaattgta tttgttaaat atgtactaca aacttagtag tttttaaaga aattgtattt 8881 gttaaatatg tactacaaac ttagtagt //