LOCUS V024534 9352 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V024534 VERSION V024534 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..4048 /label="Ager(NM_007425)" /note="Ager(NM_007425)" /gene="Ager" CDS 4055..4120 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 4140..4639 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 4660..5259 /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 5273..5861 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6068..6701 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(6830..6846) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6854..6870 /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(6878..6908) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6923..6944 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7232..7820) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7991..8851) /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(8852..8956) /gene="bla" /label="AmpR promoter" polyA_signal 9004..9138 /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 ccatgccagc ggggacagca gctagagcct gggtgctggt 2881 tcttgctcta tggggagctg tagctggtgg tcagaacatc acagcccgga ttggagagcc 2941 acttgtgcta agctgtaagg gggcccctaa gaagccgccc cagcagctag aatggaaact 3001 gaacacagga agaactgaag cttggaaggt cctctctccc cagggaggcc cctgggacag 3061 cgtggctcga atcctcccca atggttccct cctccttcca gccactggaa ttgtcgatga 3121 ggggactttc cggtgtcggg caactaacag gcgagggaag gaggtcaagt ccaactaccg 3181 agtccgagtc taccagattc ctgggaagcc agaaattgtg gatcctgcct ctgaactcac 3241 agccagtgtc cctaataagg tggggacatg tgtgtctgag ggaagctacc ctgcagggac 3301 ccttagctgg cacttagatg ggaaacttct gattcccgat ggcaaagaaa cactcgtgaa 3361 ggaagagacc aggagacacc ctgagacggg actctttaca ctgcggtcag agctgacagt 3421 gatccccacc caaggaggaa cccatcctac cttctcctgc agtttcagcc tgggccttcc 3481 ccggcgcaga cccctgaaca cagcccccat ccaactccga gtcagggagc ctgggcctcc 3541 agagggcatt cagctgttgg ttgagcctga aggtggaata gtcgctcctg gtgggactgt 3601 gaccttgacc tgtgccatct ctgcccagcc ccctcctcag gtccactgga taaaggatgg 3661 tgcacccttg cccctggctc ccagccctgt gctgctcctc cctgaggtgg ggcacgagga 3721 tgagggcacc tatagctgcg tggccaccca ccctagccac ggacctcagg aaagccctcc 3781 tgtcagcatc agggtcacag aaaccggcga tgaggggcca gctgaaggct ctgtgggtga 3841 gtctgggctg ggtacgctag ccctggcctt ggggatcctg ggaggcctgg gagtagtagc 3901 cctgctcgtc ggggctatcc tgtggcgaaa acgacaaccc aggcgtgagg agaggaaggc 3961 cccggaaagc caggaggatg aggaggaacg tgcagagctg aatcagtcag aggaagcgga 4021 gatgccagag aatggtgccg ggggaccggg atccgactac aaagaccatg acggtgatta 4081 taaagatcat gacatcgact acaaggatga cgatgacaag tagtctagat aattctaccg 4141 ggtaggggag gcgcttttcc caaggcagtc tggagcatgc gctttagcag ccccgctggg 4201 cacttggcgc tacacaagtg gcctctggcc tcgcacacat tccacatcca ccggtaggcg 4261 ccaaccggct ccgttctttg gtggcccctt cgcgccacct tctactcctc ccctagtcag 4321 gaagttcccc cccgccccgc agctcgcgtc gtgcaggacg tgacaaatgg aagtagcacg 4381 tctcactagt ctcgtgcaga tggacagcac cgctgagcaa tggaagcggg taggcctttg 4441 gggcagcggc caatagcagc tttgctcctt cgctttctgg gctcagaggc tgggaagggg 4501 tgggtccggg ggcgggctca ggggcgggct caggggcggg gcgggcgccc gaaggtcctc 4561 cggaggcccg gcattctgca cgcttcaaaa gcgcacgtct gccgcgctgt tctcctcttc 4621 ctcatctccg ggcctttcga cctgcagccc aagcttacca tgaccgagta caagcccacg 4681 gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc 4741 gccgactacc ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc 4801 gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg 4861 gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg 4921 ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa 4981 cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc 5041 gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga 5101 gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac 5161 ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga 5221 ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgac cgcgtctgga acaatcaacc 5281 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac 5341 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt 5401 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt 5461 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg 5521 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac 5581 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac 5641 tgacaattcc gtggtgttgt cggggaagct gacgtccttt ccatggctgc tcgcctgtgt 5701 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc 5761 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg 5821 ccctcagacg agtcggatct ccctttgggc cgcctccccg cctggaatta attctgcagt 5881 cgagacctag aaaaacatgg agcaatcaca agtagcaata cagcagctac caatgctgat 5941 tgtgcctggc tagaagcaca agaggaggag gaggtgggtt ttccagtcac acctcaggta 6001 cctttaagac caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag 6061 aggggactgg aagggctaat tcactcccaa cgaagacaag atatccttga tctgtggatc 6121 taccacacac aaggctactt ccctgattag cagaactaca caccagggcc aggggtcaga 6181 tatccactga cctttggatg gtgctacaag ctagtaccag ttgagccaga taaggtagaa 6241 gaggccaata aaggagagaa caccagcttg ttacaccctg tgagcctgca tgggatggat 6301 gacccggaga gagaagtgtt agagtggagg tttgacagcc gcctagcatt tcatcacgtg 6361 gcccgagagc tgcatccgga gtacttcaag aactgctgat atcgagcttg ctacaaggga 6421 ctttccgctg gggactttcc agggaggcgt ggcctgggcg ggactgggga gtggcgagcc 6481 ctcagatcct gcatataagc agctgctttt tgcctgtact gggtctctct ggttagacca 6541 gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag 6601 cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag 6661 atccctcaga cccttttagt cagtgtggaa aatctctagc agtagtagtt catgtcatct 6721 tattattcag tatttataac ttgcaaagaa atgaatatca gagagtgaga ggccttgaca 6781 ttgctagcgt tttaccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt 6841 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa 6901 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac 6961 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg 7021 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc 7081 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat 7141 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca 7201 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc 7261 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc 7321 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg 7381 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta 7441 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg 7501 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac 7561 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag 7621 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat 7681 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat 7741 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc 7801 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt 7861 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct 7921 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt 7981 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc 8041 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac 8101 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat 8161 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg 8221 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata 8281 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta 8341 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt 8401 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag 8461 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa 8521 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc 8581 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt 8641 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc 8701 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta 8761 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa 8821 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca 8881 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac 8941 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag 9001 atcaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc 9061 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta 9121 tcttatcatg tctggatcaa ctggataact caagctaacc aaaatcatcc caaacttccc 9181 accccatacc ctattaccac tgccaattac ctgtggtttc atttactcta aacctgtgat 9241 tcctctgaat tattttcatt ttaaagaaat tgtatttgtt aaatatgtac tacaaactta 9301 gtagttttta aagaaattgt atttgttaaa tatgtactac aaacttagta gt //