LOCUS V044534 9940 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V044534 VERSION V044534 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers promoter 9..235 /note="Rous sarcoma virus enhancer/promoter" /label="RSV promoter" LTR 236..416 /note="truncated 5' long terminal repeat (LTR) from HIV-1" /label="5' LTR (truncated)" misc_feature 463..588 /note="packaging signal of human immunodeficiency virus type 1" /label="HIV-1 Ψ" misc_feature 1081..1314 /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." /label="RRE" CDS 1499..1543 /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 1810..1926 /note="central polypurine tract and central termination sequence of HIV-1" /label="cPPT/CTS" promoter 2130..3312 /note="strong constitutive promoter for human elongation factor EF-1α" /label="EF-1α promoter" promoter 3329..3347 /note="promoter for bacteriophage T7 RNA polymerase" /label="T7 promoter" CDS 3369..4676 /label="FBXL20(NM_032875)" /note="FBXL20(NM_032875)" /gene="FBXL20" CDS 4677..4703 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" CDS 4707..4733 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" CDS 4737..4763 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" misc_feature 4772..5345 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" /label="IRES" CDS 5379..5777 /codon_start=1 /gene="Aspergillus terreus BSD" /note="confers resistance to blasticidin" /product="blasticidin S deaminase" /transl_table=1 /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI KAIVKDSDGQPTAVGIRELLPSGYVWEG*" /label="BSD" misc_feature 5792..6380 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6467..6700 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6772..6893 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6933..7068 /note="SV40 origin of replication" /label="SV40 ori" promoter complement(7089..7107) /note="promoter for bacteriophage T7 RNA polymerase" /label="T7 promoter" primer_bind complement(7117..7133) /note="common sequencing primer, one of multiple similar variants" /label="M13 fwd" rep_origin 7275..7730 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 7756..7860 /gene="bla" /label="AmpR promoter" CDS 7861..8721 /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" rep_origin 8892..9480 /direction=RIGHT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" protein_bind 9768..9789 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" promoter 9804..9834 /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 9842..9858 /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" primer_bind 9866..9882 /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" promoter 9903..9921 /note="promoter for bacteriophage T3 RNA polymerase" /label="T3 promoter" ORIGIN 1 aagcttaatg tagtcttatg caatactctt gtagtcttgc aacatggtaa cgatgagtta 61 gcaacatgcc ttacaaggag agaaaaagca ccgtgcatgc cgattggtgg aagtaaggtg 121 gtacgatcgt gccttattag gaaggcaaca gacgggtctg acatggattg gacgaaccac 181 tgaattgccg cattgcagag atattgtatt taagtgccta gctcgataca taaacgggtc 241 tctctggtta gaccagatct gagcctggga gctctctggc taactaggga acccactgct 301 taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga 361 ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg 421 cgcccgaaca gggacttgaa agcgaaaggg aaaccagagg agctctctcg acgcaggact 481 cggcttgctg aagcgcgcac ggcaagaggc gaggggcggc gactggtgag tacgccaaaa 541 attttgacta gcggaggcta gaaggagaga gatgggtgcg agagcgtcag tattaagcgg 601 gggagaatta gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat 661 aaattaaaac atatagtatg ggcaagcagg gagctagaac gattcgcagt taatcctggc 721 ctgttagaaa catcagaagg ctgtagacaa atactgggac agctacaacc atcccttcag 781 acaggatcag aagaacttag atcattatat aatacagtag caaccctcta ttgtgtgcat 841 caaaggatag agataaaaga caccaaggaa gctttagaca agatagagga agagcaaaac 901 aaaagtaaga ccaccgcaca gcaagcggcc gctgatcttc agacctggag gaggagatat 961 gagggacaat tggagaagtg aattatataa atataaagta gtaaaaattg aaccattagg 1021 agtagcaccc accaaggcaa agagaagagt ggtgcagaga gaaaaaagag cagtgggaat 1081 aggagctttg ttccttgggt tcttgggagc agcaggaagc actatgggcg cagcgtcaat 1141 gacgctgacg gtacaggcca gacaattatt gtctggtata gtgcagcagc agaacaattt 1201 gctgagggct attgaggcgc aacagcatct gttgcaactc acagtctggg gcatcaagca 1261 gctccaggca agaatcctgg ctgtggaaag atacctaaag gatcaacagc tcctggggat 1321 ttggggttgc tctggaaaac tcatttgcac cactgctgtg ccttggaatg ctagttggag 1381 taataaatct ctggaacaga tttggaatca cacgacctgg atggagtggg acagagaaat 1441 taacaattac acaagcttaa tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa 1501 gaatgaacaa gaattattgg aattagataa atgggcaagt ttgtggaatt ggtttaacat 1561 aacaaattgg ctgtggtata taaaattatt cataatgata gtaggaggct tggtaggttt 1621 aagaatagtt tttgctgtac tttctatagt gaatagagtt aggcagggat attcaccatt 1681 atcgtttcag acccacctcc caaccccgag gggacccgac aggcccgaag gaatagaaga 1741 agaaggtgga gagagagaca gagacagatc cattcgatta gtgaacggat ctcgacggta 1801 tcggttaact tttaaaagaa aaggggggat tggggggtac agtgcagggg aaagaatagt 1861 agacataata gcaacagaca tacaaactaa agaattacaa aaacaaatta caaaattcaa 1921 aattttatcg atgcctcccc gtcaccaccc cccccaaccc gccccgaccg gagctgagag 1981 taattcatac aaaaggactc gcccctgcct tggggaatcc cagggaccgt cgttaaactc 2041 ccactaacgt agaacccaga gatcgctgcg ttcccgcccc ctcacccgcc cgctctcgtc 2101 atcactgagg tggagaagag catgcgtgag gctccggtgc ccgtcagtgg gcagagcgca 2161 catcgcccac agtccccgag aagttggggg gaggggtcgg caattgaacc ggtgcctaga 2221 gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg 2281 agggtggggg agaaccgtat ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg 2341 ggtttgccgc cagaacacag gtaagtgccg tgtgtggttc ccgcgggcct ggcctcttta 2401 cgggttatgg cccttgcgtg ccttgaatta cttccacgcc cctggctgca gtacgtgatt 2461 cttgatcccg agcttcgggt tggaagtggg tgggagagtt cgaggccttg cgcttaagga 2521 gccccttcgc ctcgtgcttg agttgaggcc tggcctgggc gctggggccg ccgcgtgcga 2581 atctggtggc accttcgcgc ctgtctcgct gctttcgata agtctctagc catttaaaat 2641 ttttgatgat atcctgcgac gctttttttc tggcaagata gtcttgtaaa tgcgggccaa 2701 gatctgcaca ctggtatttc ggtttttggg gccgcgggcg gcgacggggc ccgtgcgtcc 2761 cagcgcacat gttcggcgag gcggggcctg cgagcgcggc caccgagaat cggacggggg 2821 tagtctcaag ctggccggcc tgctctggtg cctggcctcg cgccgccgtg tatcgccccg 2881 ccctgggcgg caaggctggc ccggtcggca ccagttgcgt gagcggaaag atggccgctt 2941 cccggccctg ctgcagggag ctcaaaatgg aggacgcggc gctcgggaga gcgggcgggt 3001 gagtcaccca cacaaaggaa aagggccttt ccgtcctcag ccgtcgcttc atgtgactcc 3061 acggagtacc gggcgccgtc caggcacctc gattagttct cgagcttttg gagtacgtcg 3121 tctttaggtt ggggggaggg gttttatgcg atggagtttc cccacactga gtgggtggag 3181 actgaagtta ggccagcttg gcacttgatg taattctcct tggaatttgc cctttttgag 3241 tttggatctt ggttcattct caagcctcag acagtggttc aaagtttttt tcttccattt 3301 caggtgtcgt gaaaactacc cctctagata atacgactca ctatagggcc cgggttggat 3361 ccgccaccat gaggagggac gtgaacggag tgaccaagag caggtttgag atgttctcaa 3421 atagtgatga agctgtaatc aataaaaaac ttcccaaaga actcctgtta cggatatttt 3481 cttttctaga tgttgttacc ctgtgccgct gtgctcaggt ctccagggcc tggaatgttc 3541 tggctctgga tggcagtaac tggcagcgaa ttgacctatt tgatttccag agggatattg 3601 agggccgagt agtggagaat atttcaaaac gatgtggggg ctttttacga aagttaagtc 3661 ttcgtggatg tcttggagtg ggagacaatg cattaagaac ctttgcacaa aactgcagga 3721 acattgaagt actgaatcta aatgggtgta caaagacaac agacgctaca tgtactagcc 3781 ttagcaagtt ctgttccaaa ctcaggcacc ttgacttggc ttcctgtaca tcaataacaa 3841 acatgtctct aaaagctctg agtgagggat gtccactgtt ggagcagttg aacatttcct 3901 ggtgtgacca agtaaccaag gatggcattc aagcactagt gaggggctgt gggggtctca 3961 aggccttatt cttaaaaggc tgcacgcagc tagaagatga agctctcaag tacataggtg 4021 cacactgccc tgaactggtg actttgaact tgcagacttg cttgcaaatc acagatgaag 4081 gtctcattac tatatgcaga gggtgccata agttacaatc cctttgtgcc tctggctgct 4141 ccaacatcac agatgccatc ctgaatgctc taggtcagaa ctgcccacgg cttagaatat 4201 tggaagtggc aagatgttct caattaacag atgtgggctt taccactcta gccaggaatt 4261 gccatgaact tgaaaagatg gacctggaag agtgtgttca gataacagat agcacattaa 4321 tccaactttc tatacactgt cctcgacttc aagtattgag tctgtctcac tgtgagctga 4381 tcacagatga tggaattcgt cacctgggga atggggcctg cgcccatgac cagctggagg 4441 tgattgagct ggacaactgc ccactaatca cagatgcatc cctggagcac ttgaagagct 4501 gtcatagcct tgagcggata gaactctatg actgccagca aatcacacgg gctggaatca 4561 agagactcag gacccattta cccaatatta aagtccacgc ctacttcgca cctgtcactc 4621 cacccccatc agtagggggc agcagacagc gcttctgcag atgctgcatc atcctatacc 4681 catacgacgt acctgattat gcaacttatc cttatgatgt acctgattat gctagctatc 4741 cttatgatgt cccagactac gcataggcta gcccctctcc ctcccccccc cctaacgtta 4801 ctggccgaag ccgcttggaa taaggccggt gtgcgtttgt ctatatgtta ttttccacca 4861 tattgccgtc ttttggcaat gtgagggccc ggaaacctgg ccctgtcttc ttgacgagca 4921 ttcctagggg tctttcccct ctcgccaaag gaatgcaagg tctgttgaat gtcgtgaagg 4981 aagcagttcc tctggaagct tcttgaagac aaacaacgtc tgtagcgacc ctttgcaggc 5041 agcggaaccc cccacctggc gacaggtgcc tctgcggcca aaagccacgt gtataagata 5101 cacctgcaaa ggcggcacaa ccccagtgcc acgttgtgag ttggatagtt gtggaaagag 5161 tcaaatggct ctcctcaagc gtattcaaca aggggctgaa ggatgcccag aaggtacccc 5221 attgtatggg atctgatctg gggcctcggt acacatgctt tacatgtgtt tagtcgaggt 5281 taaaaaaacg tctaggcccc ccgaaccacg gggacgtggt tttcctttga aaaacacgat 5341 gataatatgg ccacactaga gatccaccgg tcgccaccat ggccaagcct ttgtctcaag 5401 aagaatccac cctcattgaa agagcaacgg ctacaatcaa cagcatcccc atctctgaag 5461 actacagcgt cgccagcgca gctctctcta gcgacggccg catcttcact ggtgtcaatg 5521 tatatcattt tactggggga ccttgtgcag aactcgtggt gctgggcact gctgctgctg 5581 cggcagctgg caacctgact tgtatcgtcg cgatcggaaa tgagaacagg ggcatcttga 5641 gcccctgcgg acggtgccga caggtgcttc tcgatctgca tcctgggatc aaagccatag 5701 tgaaggacag tgatggacag ccgacggcag ttgggattcg tgaattgctg ccctctggtt 5761 atgtgtggga gggctaagcg gccgcgtcga caatcaacct ctggattaca aaatttgtga 5821 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt 5881 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa 5941 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt 6001 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct 6061 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg 6121 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc 6181 ggggaagctg acgtcctttc catggctgct cgcctgtgtt gccacctgga ttctgcgcgg 6241 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct 6301 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc 6361 cctttgggcc gcctccccgc ctggaattcg agctcggtac ctttaagacc aatgacttac 6421 aaggcagctg tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt 6481 cactcccaac gaagacaaga tctgcttttt gcttgtactg ggtctctctg gttagaccag 6541 atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc 6601 ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga 6661 tccctcagac ccttttagtc agtgtggaaa atctctagca gtagtagttc atgtcatctt 6721 attattcagt atttataact tgcaaagaaa tgaatatcag agagtgagag gaacttgttt 6781 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca 6841 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc 6901 tggctctagc tatcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg 6961 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct 7021 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta gggacgtacc 7081 caattcgccc tatagtgagt cgtattacgc gcgctcactg gccgtcgttt tacaacgtcg 7141 tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc 7201 cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct 7261 gaatggcgaa tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac 7321 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc 7381 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt 7441 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg 7501 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac 7561 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta 7621 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat 7681 ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt aggtggcact 7741 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg 7801 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt 7861 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct 7921 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca 7981 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc 8041 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc 8101 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg 8161 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta 8221 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc 8281 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt 8341 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg 8401 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct 8461 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc 8521 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct 8581 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac 8641 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc 8701 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat 8761 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg 8821 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc 8881 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa 8941 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag 9001 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta 9061 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta 9121 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag 9181 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg 9241 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg 9301 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag 9361 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc 9421 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa 9481 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg 9541 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct 9601 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa 9661 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg 9721 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag 9781 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga 9841 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cgccaagcgc 9901 gcaattaacc ctcactaaag ggaacaaaag ctggagctgc //