LOCUS V036423 9845 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V036423 VERSION V036423 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers LTR 234..414 /note="truncated 5' long terminal repeat (LTR) from HIV-1" /label="5' LTR (truncated)" misc_feature 458..583 /note="packaging signal of human immunodeficiency virus type 1" /label="HIV-1 Ψ" misc_feature 1076..1309 /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." /label="RRE" CDS 1493..1537 /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 1804..1920 /note="central polypurine tract and central termination sequence of HIV-1" /label="cPPT/CTS" promoter 2010..2213 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" regulatory 2296..2305 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 2305..2370 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" CDS 2371..2649 /codon_start=1 /gene="" /gene_synonym="" /note="binds synthetic ligands such as AP21967" /product="T2098L mutant of FKBP-rapamycin binding domain of human FRAP" /protein_id="" /transl_table=1 /translation="ILWHEMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTL KETSFNQAYGRDLMEAQEWCRKYMKSGNVKDLLQAWDLYYHVFRRISK" /label="FRB*" CDS 2656..4050 /label="TRIM15(NM_033229)" /note="TRIM15(NM_033229)" /gene="TRIM15" CDS 4057..4767 /codon_start=1 /note="mammalian codon-optimized" /product="monomeric derivative of DsRed fluorescent protein (Shaner et al., 2004)" /transl_table=1 /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA EGRHSTGGMDELYK*" /label="mCherry" promoter 4815..5026 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 5039..5307 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 5334..5933 /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 5940..6528 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6602..6835 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6907..7028 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 7047..7182 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(7220..7236) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7244..7260 /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(7268..7298) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7313..7334 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7622..8210) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8381..9241) /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(9242..9346) /gene="bla" /label="AmpR promoter" primer_bind 9820..9836 /note="common sequencing primer, one of multiple similar variants" /label="M13 fwd" ORIGIN 1 acgcgtgtag tcttatgcaa tactcttgta gtcttgcaac atggtaacga tgagttagca 61 acatgcctta caaggagaga aaaagcaccg tgcatgccga ttggtggaag taaggtggta 121 cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca tggattggac gaaccactga 181 attgccgcat tgcagagata ttgtatttaa gtgcctagct cgatacaata aacgggtctc 241 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta 301 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact 361 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg 421 cccgaacagg gacctgaaag cgaaagggaa accagagctc tctcgacgca ggactcggct 481 tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt 541 gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag 601 aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt 661 aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt 721 agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg 781 atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag 841 gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag 901 taagaccacc gcacagcaag cggccactga tcttcagacc tggaggagga gatatgaggg 961 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag 1021 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag 1081 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcc tcaatgacgc 1141 tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga 1201 gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc 1261 aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg 1321 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata 1381 aatctctgga acagattgga atcacacgac ctggatggag tgggacagag aaattaacaa 1441 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga 1501 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa 1561 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat 1621 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt 1681 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg 1741 tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac ggtatcggtt 1801 aacttttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat 1861 aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaat tcaaaatttt 1921 atcgatacta gtattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca 1981 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc 2041 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga 2101 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat 2161 tgacgcaaat gggcggtagg cgtgtacggt gggaggttta tataagcaga gctcgtttag 2221 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagattct 2281 agagctagcg aattcgccac catggactac aaagaccatg acggtgatta taaagatcat 2341 gacatcgact acaaggatga cgatgacaag atcctctggc atgagatgtg gcatgaaggc 2401 ctggaagagg catctcgttt gtactttggg gaaaggaacg tgaaaggcat gtttgaggtg 2461 ctggagccct tgcatgctat gatggaacgg ggcccccaga ctctgaagga aacatccttt 2521 aatcaggcct atggtcgaga tttaatggag gcccaagagt ggtgcaggaa gtacatgaaa 2581 tcagggaatg tcaaggacct cctccaagcc tgggacctct attatcatgt gttccgacga 2641 atctcaaagg gatccatgcc cgcaaccccg tccctgaagg tggtccatga gctgcctgcc 2701 tgtaccctct gtgcggggcc gctggaggat gcggtgacca ttccctgtgg acacaccttc 2761 tgccggctct gcctccccgc gctctcccag atgggggccc aatcctcggg caagatcctg 2821 ctctgcccgc tctgccaaga ggaggagcag gcagagactc ccatggcccc tgtgcccctg 2881 ggcccgctgg gagaaactta ctgcgaggag cacggcgaga agatctactt cttctgcgag 2941 aacgatgccg agttcctctg tgtgttctgc agggagggtc ccacgcacca ggcgcacacc 3001 gtggggttcc tggacgaggc cattcagccc taccgggatc gtctcaggag tcgactggaa 3061 gctctgagca cggagagaga tgagattgag gatgtaaagt gtcaagaaga ccagaagctt 3121 caagtgctgc tgactcagat cgaaagcaag aagcatcagg tggaaacagc ttttgagagg 3181 ctgcagcagg agctggagca gcagcgatgt ctcctgctgg ccaggctgag ggagctggag 3241 cagcagattt ggaaggagag ggatgaatat atcacaaagg tctctgagga agtcacccgg 3301 cttggagccc aggtcaagga gctggaggag aagtgtcagc agccagcaag tgagcttcta 3361 caagatgtca gagtcaacca gagcaggtgt gagatgaaga cttttgtgag tcctgaggcc 3421 atttctcctg accttgtcaa gaagatccgt gatttccaca ggaaaatact caccctccca 3481 gagatgatga ggatgttctc agaaaacttg gcgcatcatc tggaaataga ttcaggggtc 3541 atcactctgg accctcagac cgccagccgg agcctggttc tctcggaaga caggaagtca 3601 gtgaggtaca cccggcagaa gaagagcctg ccagacagcc ccctgcgctt cgacggcctc 3661 ccggcggttc tgggcttccc gggcttctcc tccgggcgcc accgctggca ggttgacctg 3721 cagctgggcg acggcggcgg ctgcacggtg ggggtggccg gggagggggt gaggaggaag 3781 ggagagatgg gactcagcgc cgaggacggc gtctgggccg tgatcatctc gcaccagcag 3841 tgctgggcca gcacctcccc gggcaccgac ctgccgctga gcgagatccc gcgcggcgtg 3901 agagtcgccc tggactacga ggcggggcag gtgaccctcc acaacgccca gacccaggag 3961 cccatcttca ccttcactgc ctctttctcc ggcaaagtct tccctttctt tgccgtctgg 4021 aaaaaaggtt cctgccttac gctgaaaggc ggatccatgg tgagcaaggg cgaggaggat 4081 aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc acatggaggg ctccgtgaac 4141 ggccacgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggg cacccagacc 4201 gccaagctga aggtgaccaa gggtggcccc ctgcccttcg cctgggacat cctgtcccct 4261 cagttcatgt acggctccaa ggcctacgtg aagcaccccg ccgacatccc cgactacttg 4321 aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc 4381 gtggtgaccg tgacccagga ctcctccctg caggacggcg agttcatcta caaggtgaag 4441 ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc agaagaagac catgggctgg 4501 gaggcctcct ccgagcggat gtaccccgag gacggcgccc tgaagggcga gatcaagcag 4561 aggctgaagc tgaaggacgg cggccactac gacgctgagg tcaagaccac ctacaaggcc 4621 aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca tcaagttgga catcacctcc 4681 cacaacgagg actacaccat cgtggaacag tacgaacgcg ccgagggccg ccactccacc 4741 ggcggcatgg acgagctgta caagtgagga tccgcggccg cgaaggatct gcgatcgctc 4801 cggtgcccgt cagtgggcag agcgcacatc gcccacagtc cccgagaagt tggggggagg 4861 ggtcggcaat tgaacgggtg cctagagaag gtggcgcggg gtaaactggg aaagtgatgt 4921 cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt 4981 cgccgtgaac gttctttttc gcaacgggtt tgccgccaga acacagctga agcttcgagg 5041 ggctcgcatc tctccttcac gcgcccgccg ccctacctga ggccgccatc cacgccggtt 5101 gagtcgcgtt ctgccgcctc ccgcctgtgg tgcctcctga actgcgtccg ccgtctaggt 5161 aagtttaaag ctcaggtcga gaccgggcct ttgtccggcg ctcccttgga gcctacctag 5221 actcagccgg ctctccacgc tttgcctgac cctgcttgct caactctacg tctttgtttc 5281 gttttctgtt ctgcgccgtt acagatccaa gctgtgaccg gcgcctacgc tagatgaccg 5341 agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt cccccgggcc gtacgcaccc 5401 tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgacccg gaccgccaca 5461 tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca 5521 aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg 5581 aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc 5641 tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt 5701 ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg 5761 tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct 5821 ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg 5881 aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tgagtcgaca 5941 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc 6001 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta 6061 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt 6121 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg 6181 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta 6241 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt 6301 tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg 6361 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca 6421 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc 6481 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct ggtaccttta 6541 agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga 6601 ctggaagggc taattcactc ccaacgaaaa taagatctgc tttttgcttg tactgggtct 6661 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt 6721 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac 6781 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtagt 6841 agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat atcagagagt 6901 gagaggaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat 6961 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat 7021 gtatcttatc atgtctggct ctagctatcc cgcccctaac tccgcccagt tccgcccatt 7081 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct 7141 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctagacttt tgcagagacg 7201 gcccaaattc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa 7261 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga 7321 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt 7381 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct 7441 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat 7501 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga 7561 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt 7621 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt 7681 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc 7741 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa 7801 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct 7861 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta 7921 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg 7981 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc 8041 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta 8101 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg 8161 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt 8221 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg 8281 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta 8341 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg 8401 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg 8461 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc 8521 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg 8581 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg 8641 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag 8701 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat 8761 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc 8821 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc 8881 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa 8941 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac 9001 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt 9061 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc 9121 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa 9181 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca 9241 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat 9301 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa 9361 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc 9421 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca 9481 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc 9541 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag 9601 agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga 9661 gaaaataccg catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat 9721 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat 9781 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgcca 9841 agctg //