LOCUS V020142 10928 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V020142 VERSION V020142 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..5654 /label="EGFR(NM_001346941)" /note="EGFR(NM_001346941)" /gene="EGFR" CDS 5661..5684 /codon_start=1 /product="FLAG® epitope tag, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDDDDK" /label="FLAG" promoter 5716..6215 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 6236..6835 /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 6849..7437 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 7644..8277 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(8406..8422) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 8430..8446 /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(8454..8484) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 8499..8520 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(8808..9396) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(9567..10427) /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(10428..10532) /gene="bla" /label="AmpR promoter" polyA_signal 10580..10714 /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 ccaccatgcg accctccggg acggccgggg cagcgctcct ggcgctgctg gctgcgctct 2881 gcccggcgag tcgggctctg gaggaaaaga aaggtaatta tgtggtgaca gatcacggct 2941 cgtgcgtccg agcctgtggg gccgacagct atgagatgga ggaagacggc gtccgcaagt 3001 gtaagaagtg cgaagggcct tgccgcaaag tgtgtaacgg aataggtatt ggtgaattta 3061 aagactcact ctccataaat gctacgaata ttaaacactt caaaaactgc acctccatca 3121 gtggcgatct ccacatcctg ccggtggcat ttaggggtga ctccttcaca catactcctc 3181 ctctggatcc acaggaactg gatattctga aaaccgtaaa ggaaatcaca gggtttttgc 3241 tgattcaggc ttggcctgaa aacaggacgg acctccatgc ctttgagaac ctagaaatca 3301 tacgcggcag gaccaagcaa catggtcagt tttctcttgc agtcgtcagc ctgaacataa 3361 catccttggg attacgctcc ctcaaggaga taagtgatgg agatgtgata atttcaggaa 3421 acaaaaattt gtgctatgca aatacaataa actggaaaaa actgtttggg acctccggtc 3481 agaaaaccaa aattataagc aacagaggtg aaaacagctg caaggccaca ggccaggtct 3541 gccatgcctt gtgctccccc gagggctgct ggggcccgga gcccagggac tgcgtctctt 3601 gccggaatgt cagccgaggc agggaatgcg tggacaagtg caaccttctg gagggtgagc 3661 caagggagtt tgtggagaac tctgagtgca tacagtgcca cccagagtgc ctgcctcagg 3721 ccatgaacat cacctgcaca ggacggggac cagacaactg tatccagtgt gcccactaca 3781 ttgacggccc ccactgcgtc aagacctgcc cggcaggagt catgggagaa aacaacaccc 3841 tggtctggaa gtacgcagac gccggccatg tgtgccacct gtgccatcca aactgcacct 3901 acggatgcac tgggccaggt cttgaaggct gtccaacgaa tgggcctaag atcccgtcca 3961 tcgccactgg gatggtgggg gccctcctct tgctgctggt ggtggccctg gggatcggcc 4021 tcttcatgcg aaggcgccac atcgttcgga agcgcacgct gcggaggctg ctgcaggaga 4081 gggagcttgt ggagcctctt acacccagtg gagaagctcc caaccaagct ctcttgagga 4141 tcttgaagga aactgaattc aaaaagatca aagtgctggg ctccggtgcg ttcggcacgg 4201 tgtataaggg actctggatc ccagaaggtg agaaagttaa aattcccgtc gctatcaagg 4261 aattaagaga agcaacatct ccgaaagcca acaaggaaat cctcgatgaa gcctacgtga 4321 tggccagcgt ggacaacccc cacgtgtgcc gcctgctggg catctgcctc acctccaccg 4381 tgcagctcat cacgcagctc atgcccttcg gctgcctcct ggactatgtc cgggaacaca 4441 aagacaatat tggctcccag tacctgctca actggtgtgt gcagatcgca aagggcatga 4501 actacttgga ggaccgtcgc ttggtgcacc gcgacctggc agccaggaac gtactggtga 4561 aaacaccgca gcatgtcaag atcacagatt ttgggctggc caaactgctg ggtgcggaag 4621 agaaagaata ccatgcagaa ggaggcaaag tgcctatcaa gtggatggca ttggaatcaa 4681 ttttacacag aatctatacc caccagagtg atgtctggag ctacggggtg actgtttggg 4741 agttgatgac ctttggatcc aagccatatg acggaatccc tgccagcgag atctcctcca 4801 tcctggagaa aggagaacgc ctccctcagc cacccatatg taccatcgat gtctacatga 4861 tcatggtcaa gtgctggatg atagacgcag atagtcgccc aaagttccgt gagttgatca 4921 tcgaattctc caaaatggcc cgagaccccc agcgctacct tgtcattcag ggggatgaaa 4981 gaatgcattt gccaagtcct acagactcca acttctaccg tgccctgatg gatgaagaag 5041 acatggacga cgtggtggat gccgacgagt acctcatccc acagcagggc ttcttcagca 5101 gcccctccac gtcacggact cccctcctga gctctctgag tgcaaccagc aacaattcca 5161 ccgtggcttg cattgataga aatgggctgc aaagctgtcc catcaaggaa gacagcttct 5221 tgcagcgata cagctcagac cccacaggcg ccttgactga ggacagcata gacgacacct 5281 tcctcccagt gcctgaatac ataaaccagt ccgttcccaa aaggcccgct ggctctgtgc 5341 agaatcctgt ctatcacaat cagcctctga accccgcgcc cagcagagac ccacactacc 5401 aggaccccca cagcactgca gtgggcaacc ccgagtatct caacactgtc cagcccacct 5461 gtgtcaacag cacattcgac agccctgccc actgggccca gaaaggcagc caccaaatta 5521 gcctggacaa ccctgactac cagcaggact tctttcccaa ggaagccaag ccaaatggca 5581 tctttaaggg ctccacagct gaaaatgcag aatacctaag ggtcgcgcca caaagcagtg 5641 aatttattgg agcagaattc gattacaagg atgacgacga taagtgagga tcccgcgact 5701 ctagataatt ctaccgggta ggggaggcgc ttttcccaag gcagtctgga gcatgcgctt 5761 tagcagcccc gctgggcact tggcgctaca caagtggcct ctggcctcgc acacattcca 5821 catccaccgg taggcgccaa ccggctccgt tctttggtgg ccccttcgcg ccaccttcta 5881 ctcctcccct agtcaggaag ttcccccccg ccccgcagct cgcgtcgtgc aggacgtgac 5941 aaatggaagt agcacgtctc actagtctcg tgcagatgga cagcaccgct gagcaatgga 6001 agcgggtagg cctttggggc agcggccaat agcagctttg ctccttcgct ttctgggctc 6061 agaggctggg aaggggtggg tccgggggcg ggctcagggg cgggctcagg ggcggggcgg 6121 gcgcccgaag gtcctccgga ggcccggcat tctgcacgct tcaaaagcgc acgtctgccg 6181 cgctgttctc ctcttcctca tctccgggcc tttcgacctg cagcccaagc ttaccatgac 6241 cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac 6301 cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca 6361 catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg 6421 caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt 6481 cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg 6541 gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc 6601 gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc 6661 cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac 6721 ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt 6781 cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgaccgcg 6841 tctggaacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact 6901 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg 6961 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg 7021 aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa 7081 cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc 7141 ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg 7201 ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaagctgacg tcctttccat 7261 ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt 7321 cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc 7381 cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg 7441 gaattaattc tgcagtcgag acctagaaaa acatggagca atcacaagta gcaatacagc 7501 agctaccaat gctgattgtg cctggctaga agcacaagag gaggaggagg tgggttttcc 7561 agtcacacct caggtacctt taagaccaat gacttacaag gcagctgtag atcttagcca 7621 ctttttaaaa gaaaagaggg gactggaagg gctaattcac tcccaacgaa gacaagatat 7681 ccttgatctg tggatctacc acacacaagg ctacttccct gattagcaga actacacacc 7741 agggccaggg gtcagatatc cactgacctt tggatggtgc tacaagctag taccagttga 7801 gccagataag gtagaagagg ccaataaagg agagaacacc agcttgttac accctgtgag 7861 cctgcatggg atggatgacc cggagagaga agtgttagag tggaggtttg acagccgcct 7921 agcatttcat cacgtggccc gagagctgca tccggagtac ttcaagaact gctgatatcg 7981 agcttgctac aagggacttt ccgctgggga ctttccaggg aggcgtggcc tgggcgggac 8041 tggggagtgg cgagccctca gatcctgcat ataagcagct gctttttgcc tgtactgggt 8101 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc 8161 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg 8221 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta 8281 gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga 8341 gtgagaggcc ttgacattgc tagcgtttta ccgtcgacct ctagctagag cttggcgtaa 8401 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata 8461 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta 8521 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa 8581 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg 8641 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag 8701 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa 8761 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc 8821 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca 8881 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg 8941 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct 9001 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt 9061 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag 9121 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc 9181 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac 9241 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga 9301 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc 9361 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg 9421 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca 9481 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt 9541 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca 9601 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg 9661 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca 9721 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt 9781 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt 9841 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca 9901 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca 9961 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga 10021 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact 10081 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga 10141 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg 10201 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc 10261 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga 10321 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat 10381 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt 10441 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt 10501 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac 10561 gtcgacggat cgggagatca acttgtttat tgcagcttat aatggttaca aataaagcaa 10621 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc 10681 caaactcatc aatgtatctt atcatgtctg gatcaactgg ataactcaag ctaaccaaaa 10741 tcatcccaaa cttcccaccc cataccctat taccactgcc aattacctgt ggtttcattt 10801 actctaaacc tgtgattcct ctgaattatt ttcattttaa agaaattgta tttgttaaat 10861 atgtactaca aacttagtag tttttaaaga aattgtattt gttaaatatg tactacaaac 10921 ttagtagt //