LOCUS V024700 9866 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V024700 VERSION V024700 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" promoter 2337..3519 /note="strong constitutive promoter for human elongation factor EF-1α" /label="EF-1α promoter" CDS 3551..3568 /codon_start=1 /product="6xHis affinity tag" /transl_table=1 /translation="HHHHHH" /label="6xHis" CDS 3569..4516 /label="OR2H1(NM_001318014)" /note="OR2H1(NM_001318014)" /gene="OR2H1" CDS 4523..4588 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 4616..5202 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" /label="IRES2" CDS 5222..5821 /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 5835..6423 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6630..7263 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(7391..7407) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7415..7431 /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(7439..7469) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7484..7505 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7793..8381) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8552..9412) /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(9413..9517) /gene="bla" /label="AmpR promoter" polyA_signal 9565..9699 /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 cgatgagtaa ttcatacaaa aggactcgcc cctgccttgg 2221 ggaatcccag ggaccgtcgt taaactccca ctaacgtaga acccagagat cgctgcgttc 2281 ccgccccctc acccgcccgc tctcgtcatc actgaggtgg agaagagcat gcgtgaggct 2341 ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag ttggggggag 2401 gggtcggcaa ttgaaccggt gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg 2461 tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata agtgcagtag 2521 tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacaggta agtgccgtgt 2581 gtggttcccg cgggcctggc ctctttacgg gttatggccc ttgcgtgcct tgaattactt 2641 ccacgcccct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg 2701 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg 2761 cttgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct 2821 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg 2881 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc 2941 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga 3001 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct 3061 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca 3121 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg 3181 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg 3241 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat 3301 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg 3361 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa 3421 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca 3481 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gaattagctt ggtacgaatt 3541 cgccaccatg caccatcacc atcaccatat ggttaaccaa agctccccca tgggcttcct 3601 ccttctgggc ttctctgaac acccagcact ggaaaggact ctctttgtgg ttgtcttcac 3661 ttcctacctc ttgaccctgg tgggcaacac actcatcatc ctgctgtctg tactgtaccc 3721 caggctccac tctccaatgt actttttcct ctctgacctc tccttcttgg acctctgctt 3781 taccacaagt tgtgtccccc agatgctggt caacctctgg ggcccaaaga agaccatcag 3841 cttcctggga tgctctgtcc agctcttcat cttcctgtcc ctggggacca ctgagtgcat 3901 cctcctgaca gtgatggcct ttgaccgata cgtggctgtc tgccagcccc tccactatgc 3961 caccatcatc cacccccgcc tgtgctggca gctggcatct gtggcctggg ttatgagtct 4021 ggttcaatcg atagtccaga caccatccac cctccacttg cccttctgtc cccaccagca 4081 gatagatgac tttttatgtg aggtcccatc tctgattcga ctctcctgtg gagatacctc 4141 ctacaatgaa atccagttgg ctgtgtccag tgtcatcttc gtggttgtgc ctctcagcct 4201 catccttgcc tcttatggag ccactgccca ggcagtgctg aggattaact ctgccacagc 4261 atggagaaag gcctttggga cctgctcctc ccatctcact gtggtcaccc tcttctacag 4321 ctcagtcatt gctgtctacc tccagcccaa aaatccgtat gcccaaggga ggggcaagtt 4381 ctttggtctc ttctatgcag tgggcactcc ttcacttaac cctctcgtat acaccctgag 4441 gaacaaggag ataaagcgag cactcaggag gttactaggg aaggaaagag actccaggga 4501 aagctggaga gctgctcata tggactacaa agaccatgac ggtgattata aagatcatga 4561 catcgactac aaggatgacg atgacaagta ggcctgcgat atccagcgga tcccgcccct 4621 ctccctcccc cccccctaac gttactggcc gaagccgctt ggaataaggc cggtgtgcgt 4681 ttgtctatat gttattttcc accatattgc cgtcttttgg caatgtgagg gcccggaaac 4741 ctggccctgt cttcttgacg agcattccta ggggtctttc ccctctcgcc aaaggaatgc 4801 aaggtctgtt gaatgtcgtg aaggaagcag ttcctctgga agcttcttga agacaaacaa 4861 cgtctgtagc gaccctttgc aggcagcgga accccccacc tggcgacagg tgcctctgcg 4921 gccaaaagcc acgtgtataa gatacacctg caaaggcggc acaaccccag tgccacgttg 4981 tgagttggat agttgtggaa agagtcaaat ggctctcctc aagcgtattc aacaaggggc 5041 tgaaggatgc ccagaaggta ccccattgta tgggatctga tctggggcct cggtgcacat 5101 gctttacatg tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac cacggggacg 5161 tggttttcct ttgaaaaaca cgatgataag cttgccacaa cccacaagga gacgaccttc 5221 catgaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc cccgggccgt 5281 acgcaccctc gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgacccgga 5341 ccgccacatc gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga 5401 catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga 5461 gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg 5521 ttcccggctg gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga 5581 gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg 5641 cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct 5701 ggagacctcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc 5761 cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgccta 5821 gacgcgtctg gaacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc 5881 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg 5941 ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc 6001 tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg 6061 acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg 6121 ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga 6181 caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct 6241 ttccatggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg 6301 tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc 6361 ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc 6421 cgcctggaat taattctgca gtcgagacct agaaaaacat ggagcaatca caagtagcaa 6481 tacagcagct accaatgctg attgtgcctg gctagaagca caagaggagg aggaggtggg 6541 ttttccagtc acacctcagg tacctttaag accaatgact tacaaggcag ctgtagatct 6601 tagccacttt ttaaaagaaa agaggggact ggaagggcta attcactccc aacgaagaca 6661 agatatcctt gatctgtgga tctaccacac acaaggctac ttccctgatt agcagaacta 6721 cacaccaggg ccaggggtca gatatccact gacctttgga tggtgctaca agctagtacc 6781 agttgagcca gataaggtag aagaggccaa taaaggagag aacaccagct tgttacaccc 6841 tgtgagcctg catgggatgg atgacccgga gagagaagtg ttagagtgga ggtttgacag 6901 ccgcctagca tttcatcacg tggcccgaga gctgcatccg gagtacttca agaactgctg 6961 atatcgagct tgctacaagg gactttccgc tggggacttt ccagggaggc gtggcctggg 7021 cgggactggg gagtggcgag ccctcagatc ctgcatataa gcagctgctt tttgcctgta 7081 ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc 7141 cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt 7201 tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta 7261 gcagtagtag ttcatgtcat cttattattc agtatttata acttgcaaag aaatgaatat 7321 cagagagtga gaggccttga cattgctagc gtttaccgtc gacctctagc tagagcttgg 7381 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca 7441 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca 7501 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc 7561 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt 7621 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact 7681 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag 7741 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata 7801 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc 7861 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg 7921 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc 7981 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg 8041 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc 8101 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga 8161 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg 8221 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa 8281 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg 8341 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt 8401 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat 8461 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct 8521 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta 8581 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa 8641 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac 8701 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa 8761 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag 8821 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg 8881 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag 8941 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg 9001 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc 9061 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat 9121 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata 9181 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa 9241 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca 9301 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc 9361 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc 9421 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg 9481 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac 9541 ctgacgtcga cggatcggga gatcaacttg tttattgcag cttataatgg ttacaaataa 9601 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt 9661 ttgtccaaac tcatcaatgt atcttatcat gtctggatca actggataac tcaagctaac 9721 caaaatcatc ccaaacttcc caccccatac cctattacca ctgccaatta cctgtggttt 9781 catttactct aaacctgtga ttcctctgaa ttattttcat tttaaagaaa ttgtatttgt 9841 taaatatgta ctacaaactt agtagt //