LOCUS V025069 10672 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V025069 VERSION V025069 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..5368 /label="Tas1r1(NM_031867)" /note="Tas1r1(NM_031867)" /gene="Tas1r1" CDS 5375..5440 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 5460..5959 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 5980..6579 /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 6593..7181 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 7388..8021 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(8150..8166) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 8174..8190 /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(8198..8228) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 8243..8264 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(8552..9140) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(9311..10171) /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(10172..10276) /gene="bla" /label="AmpR promoter" polyA_signal 10324..10458 /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 ccatgctttt ctgggcagct cacctgctgc tcagcctgca 2881 gctggccgtt gcttactgct gggctttcag ctgccaaagg acagaatcct ctccaggttt 2941 cagcctccct ggggacttcc tcctggcagg cctgttctcc ctccatgctg actgtctgca 3001 ggtgagacac agacctctgg tgacaagttg tgacaggtct gacagcttca acggccatgg 3061 ctatcacctc ttccaagcca tgcggttcac cgttgaggag ataaacaact ccacagctct 3121 gcttcccaac atcaccctgg ggtatgaact gtatgacgtg tgctcagagt cttccaatgt 3181 ctatgccacc ctgagggtgc tcgcccagca agggacaggc cacctagaga tgcagagaga 3241 tcttcgcaac cactcctcca aggtggtggc actcattggg cctgataaca ctgaccacgc 3301 tgtcaccact gctgccctgc tgagcccttt tctgatgccc ctggtcagct atgaggcgag 3361 cagcgtgatc ctcagtggga agcgcaagtt cccgtccttc ttgcgcacca tccccagcga 3421 taagtaccag gtggaagtca tagtgcggct gctgcagagc ttcggctggg tctggatctc 3481 gctcgttggc agctatggtg actacgggca gctgggcgta caggcgctgg aggagctggc 3541 cactccacgg ggcatctgcg tcgccttcaa ggacgtggtg cctctctccg cccaggcggg 3601 tgacccaagg atgcagcgca tgatgctgcg tctggctcga gccaggacca ccgtggtcgt 3661 ggtcttctct aaccggcacc tggctggagt gttcttcagg tctgtggtgc tggccaacct 3721 gactggcaaa gtgtggatcg cctccgaaga ctgggccatc tccacgtaca tcaccaatgt 3781 gcccgggatc cagggcattg ggacggtgct gggggtggcc atccagcaga gacaagtccc 3841 tggcctgaag gagtttgaag agtcctatgt ccaggcagtg atgggtgctc ccagaacttg 3901 cccagagggg tcctggtgcg gcactaacca gctgtgcagg gagtgtcacg ctttcacgac 3961 atggaacatg cccgagcttg gagccttctc catgagcgct gcctacaatg tgtatgaggc 4021 tgtgtatgct gtggcccacg gcctccacca gctcctggga tgtacctctg ggacctgtgc 4081 cagaggccca gtctacccct ggcagcttct tcagcagatc tacaaggtga atttccttct 4141 acataagaag actgtagcat tcgatgacaa gggggaccct ctaggttatt atgacatcat 4201 cgcctgggac tggaatggac ctgaatggac ctttgaggtc attggttctg cctcactgtc 4261 tccagttcat ctagacataa ataagacaaa aatccagtgg cacgggaaga acaatcaggt 4321 gcctgtgtca gtgtgtacca gggactgtct cgaagggcac cacaggttgg tcatgggttc 4381 ccaccactgc tgcttcgagt gcatgccctg tgaagctggg acatttctca acacgagtga 4441 gcttcacacc tgccagcctt gtggaacaga agaatgggcc cctgagggga gctcagcctg 4501 cttctcacgc accgtggagt tcttggggtg gcatgaaccc atctctttgg tgctattagc 4561 agctaacacg ctattgctgc tgctgctgat tgggactgct ggcctgtttg cctggcgtct 4621 tcacacgcct gttgtgaggt cagctggggg taggctgtgc ttcctcatgc tgggttcctt 4681 ggtagctggg agttgcagcc tctacagctt cttcgggaag cccacggtgc ccgcgtgctt 4741 gctgcgtcag cccctctttt ctctcgggtt tgccattttc ctctcctgtc tgacaatccg 4801 ctccttccaa ctggtcatca tcttcaagtt ttctaccaag gtacccacat tctaccacac 4861 ttgggcccaa aaccatggtg ccggaatatt cgtcattgtc agctccacgg tccatttgtt 4921 cctctgtctc acgtggcttg caatgtggac cccacggccc accagggagt accagcgctt 4981 cccccatctg gtgattcttg agtgcacaga ggtcaactct gtgggcttcc tggtggcttt 5041 cgcacacaac atcctcctct ccatcagcac ctttgtctgc agctacctgg gtaaggaact 5101 gccggagaac tataacgaag ccaaatgtgt caccttcagc ctgctcctcc acttcgtatc 5161 ctggatcgct ttcttcacca tgtccagcat ttaccagggc agctacctac ccgcggtcaa 5221 tgtgctggca gggctggcca ctctgagtgg cggcttcagc ggctatttcc tccctaaatg 5281 ctacgtgatt ctctgccgtc cagaactcaa caacacagaa cactttcagg cctccatcca 5341 ggactacacg aggcgctgcg gcactaccgg atccgactac aaagaccatg acggtgatta 5401 taaagatcat gacatcgact acaaggatga cgatgacaag tagtctagat aattctaccg 5461 ggtaggggag gcgcttttcc caaggcagtc tggagcatgc gctttagcag ccccgctggg 5521 cacttggcgc tacacaagtg gcctctggcc tcgcacacat tccacatcca ccggtaggcg 5581 ccaaccggct ccgttctttg gtggcccctt cgcgccacct tctactcctc ccctagtcag 5641 gaagttcccc cccgccccgc agctcgcgtc gtgcaggacg tgacaaatgg aagtagcacg 5701 tctcactagt ctcgtgcaga tggacagcac cgctgagcaa tggaagcggg taggcctttg 5761 gggcagcggc caatagcagc tttgctcctt cgctttctgg gctcagaggc tgggaagggg 5821 tgggtccggg ggcgggctca ggggcgggct caggggcggg gcgggcgccc gaaggtcctc 5881 cggaggcccg gcattctgca cgcttcaaaa gcgcacgtct gccgcgctgt tctcctcttc 5941 ctcatctccg ggcctttcga cctgcagccc aagcttacca tgaccgagta caagcccacg 6001 gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc 6061 gccgactacc ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc 6121 gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg 6181 gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg 6241 ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa 6301 cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc 6361 gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga 6421 gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac 6481 ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga 6541 ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgac cgcgtctgga acaatcaacc 6601 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac 6661 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt 6721 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt 6781 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg 6841 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac 6901 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac 6961 tgacaattcc gtggtgttgt cggggaagct gacgtccttt ccatggctgc tcgcctgtgt 7021 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc 7081 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg 7141 ccctcagacg agtcggatct ccctttgggc cgcctccccg cctggaatta attctgcagt 7201 cgagacctag aaaaacatgg agcaatcaca agtagcaata cagcagctac caatgctgat 7261 tgtgcctggc tagaagcaca agaggaggag gaggtgggtt ttccagtcac acctcaggta 7321 cctttaagac caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag 7381 aggggactgg aagggctaat tcactcccaa cgaagacaag atatccttga tctgtggatc 7441 taccacacac aaggctactt ccctgattag cagaactaca caccagggcc aggggtcaga 7501 tatccactga cctttggatg gtgctacaag ctagtaccag ttgagccaga taaggtagaa 7561 gaggccaata aaggagagaa caccagcttg ttacaccctg tgagcctgca tgggatggat 7621 gacccggaga gagaagtgtt agagtggagg tttgacagcc gcctagcatt tcatcacgtg 7681 gcccgagagc tgcatccgga gtacttcaag aactgctgat atcgagcttg ctacaaggga 7741 ctttccgctg gggactttcc agggaggcgt ggcctgggcg ggactgggga gtggcgagcc 7801 ctcagatcct gcatataagc agctgctttt tgcctgtact gggtctctct ggttagacca 7861 gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag 7921 cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag 7981 atccctcaga cccttttagt cagtgtggaa aatctctagc agtagtagtt catgtcatct 8041 tattattcag tatttataac ttgcaaagaa atgaatatca gagagtgaga ggccttgaca 8101 ttgctagcgt tttaccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt 8161 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa 8221 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac 8281 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg 8341 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc 8401 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat 8461 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca 8521 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc 8581 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc 8641 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg 8701 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta 8761 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg 8821 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac 8881 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag 8941 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat 9001 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat 9061 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc 9121 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt 9181 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct 9241 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt 9301 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc 9361 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac 9421 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat 9481 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg 9541 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata 9601 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta 9661 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt 9721 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag 9781 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa 9841 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc 9901 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt 9961 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc 10021 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta 10081 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa 10141 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca 10201 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac 10261 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag 10321 atcaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc 10381 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta 10441 tcttatcatg tctggatcaa ctggataact caagctaacc aaaatcatcc caaacttccc 10501 accccatacc ctattaccac tgccaattac ctgtggtttc atttactcta aacctgtgat 10561 tcctctgaat tattttcatt ttaaagaaat tgtatttgtt aaatatgtac tacaaactta 10621 gtagttttta aagaaattgt atttgttaaa tatgtactac aaacttagta gt //