LOCUS V020128 9274 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V020128 VERSION V020128 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..4004 /label="MAP2K1(NM_002755)" /note="MAP2K1(NM_002755)" /gene="MAP2K1" CDS 4005..4031 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" promoter 4062..4561 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 4582..5181 /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 5195..5783 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5990..6623 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(6752..6768) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6776..6792 /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(6800..6830) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6845..6866 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7154..7742) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7913..8773) /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(8774..8878) /gene="bla" /label="AmpR promoter" polyA_signal 8926..9060 /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 ccaccatgcc caagaagaag ccgacgccca tccagctgaa cccggccccc gacggctctg 2881 cagttaacgg gaccagctct gcggagacca acttggaggc cttgcagaag aagctggagg 2941 agctagagct tgatgagcag cagcgaaagc gccttgaggc ctttcttacc cagaagcaga 3001 aggtgggaga actgaaggat gacgactttg agaagatcag tgagctgggg gctggcaatg 3061 gcggtgtggt gttcaaggtc tcccacaagc cttctggcct ggtcatggcc agaaagctaa 3121 ttcatctgga gatcaaaccc gcaatccgga accagatcat aagggagctg caggttctgc 3181 atgagtgcaa ctctccgtac atcgtgggct tctatggtgc gttctacagc gatggcgaga 3241 tcagtatctg catggagcac atggatggag gttctctgga tcaagtcctg aagaaagctg 3301 gaagaattcc tgaacaaatt ttaggaaaag ttagcattgc tgtaataaaa ggcctgacat 3361 atctgaggga gaagcacaag atcatgcaca gagatgtcaa gccctccaac atcctagtca 3421 actcccgtgg ggagatcaag ctctgtgact ttggggtcag cgggcagctc atcgactcca 3481 tggccaactc cttcgtgggc acaaggtcct acatgtcgcc agaaagactc caggggactc 3541 attactctgt gcagtcagac atctggagca tgggactgtc tctggtagag atggcggttg 3601 ggaggtatcc catccctcct ccagatgcca aggagctgga gctgatgttt gggtgccagg 3661 tggaaggaga tgcggctgag accccaccca ggccaaggac ccccgggagg ccccttagct 3721 catacggaat ggacagccga cctcccatgg caatttttga gttgttggat tacatagtca 3781 acgagcctcc tccaaaactg cccagtggag tgttcagtct ggaatttcaa gattttgtga 3841 ataaatgctt aataaaaaac cccgcagaga gagcagattt gaagcaactc atggttcatg 3901 cttttatcaa gagatctgat gctgaggaag tggattttgc aggttggctc tgctccacca 3961 tcggccttaa ccagcccagc acaccaaccc atgctgctgg cgtctaccca tacgacgtac 4021 cagattacgc ttaggatccc gcgactctag ataattctac cgggtagggg aggcgctttt 4081 cccaaggcag tctggagcat gcgctttagc agccccgctg ggcacttggc gctacacaag 4141 tggcctctgg cctcgcacac attccacatc caccggtagg cgccaaccgg ctccgttctt 4201 tggtggcccc ttcgcgccac cttctactcc tcccctagtc aggaagttcc cccccgcccc 4261 gcagctcgcg tcgtgcagga cgtgacaaat ggaagtagca cgtctcacta gtctcgtgca 4321 gatggacagc accgctgagc aatggaagcg ggtaggcctt tggggcagcg gccaatagca 4381 gctttgctcc ttcgctttct gggctcagag gctgggaagg ggtgggtccg ggggcgggct 4441 caggggcggg ctcaggggcg gggcgggcgc ccgaaggtcc tccggaggcc cggcattctg 4501 cacgcttcaa aagcgcacgt ctgccgcgct gttctcctct tcctcatctc cgggcctttc 4561 gacctgcagc ccaagcttac catgaccgag tacaagccca cggtgcgcct cgccacccgc 4621 gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg 4681 cgccacaccg tcgatccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc 4741 ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg 4801 gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg 4861 cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga aggcctcctg 4921 gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac 4981 caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc 5041 gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg 5101 ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg 5161 acccgcaagc ccggtgcctg accgcgtctg gaacaatcaa cctctggatt acaaaatttg 5221 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc 5281 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta 5341 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt 5401 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca 5461 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc 5521 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt 5581 gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct ggattctgcg 5641 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg 5701 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat 5761 ctccctttgg gccgcctccc cgcctggaat taattctgca gtcgagacct agaaaaacat 5821 ggagcaatca caagtagcaa tacagcagct accaatgctg attgtgcctg gctagaagca 5881 caagaggagg aggaggtggg ttttccagtc acacctcagg tacctttaag accaatgact 5941 tacaaggcag ctgtagatct tagccacttt ttaaaagaaa agaggggact ggaagggcta 6001 attcactccc aacgaagaca agatatcctt gatctgtgga tctaccacac acaaggctac 6061 ttccctgatt agcagaacta cacaccaggg ccaggggtca gatatccact gacctttgga 6121 tggtgctaca agctagtacc agttgagcca gataaggtag aagaggccaa taaaggagag 6181 aacaccagct tgttacaccc tgtgagcctg catgggatgg atgacccgga gagagaagtg 6241 ttagagtgga ggtttgacag ccgcctagca tttcatcacg tggcccgaga gctgcatccg 6301 gagtacttca agaactgctg atatcgagct tgctacaagg gactttccgc tggggacttt 6361 ccagggaggc gtggcctggg cgggactggg gagtggcgag ccctcagatc ctgcatataa 6421 gcagctgctt tttgcctgta ctgggtctct ctggttagac cagatctgag cctgggagct 6481 ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca 6541 agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta 6601 gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat cttattattc agtatttata 6661 acttgcaaag aaatgaatat cagagagtga gaggccttga cattgctagc gttttaccgt 6721 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt 6781 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg 6841 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg 6901 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc 6961 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc 7021 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata 7081 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg 7141 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct 7201 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa 7261 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc 7321 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt 7381 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg 7441 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg 7501 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct 7561 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc 7621 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg 7681 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc 7741 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt 7801 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa 7861 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat 7921 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct 7981 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg 8041 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag 8101 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta 8161 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg 8221 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg 8281 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct 8341 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta 8401 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg 8461 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc 8521 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg 8581 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga 8641 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg 8701 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat 8761 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc 8821 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca 8881 catttccccg aaaagtgcca cctgacgtcg acggatcggg agatcaactt gtttattgca 8941 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt 9001 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc 9061 aactggataa ctcaagctaa ccaaaatcat cccaaacttc ccaccccata ccctattacc 9121 actgccaatt acctgtggtt tcatttactc taaacctgtg attcctctga attattttca 9181 ttttaaagaa attgtatttg ttaaatatgt actacaaact tagtagtttt taaagaaatt 9241 gtatttgtta aatatgtact acaaacttag tagt //