LOCUS V039175 9786 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V039175 VERSION V039175 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..1921 /note="central polypurine tract and central termination sequence of HIV-1" /label="cPPT/CTS" promoter 2011..2214 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 2303..3868 /label="TRAF6(NM_145803)" /note="TRAF6(NM_145803)" /gene="TRAF6" CDS 3875..3940 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3984..4195 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4208..4476 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 4511..5218 /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" CDS 5228..5281 /codon_start=1 /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /product="2A peptide from Thosea asigna virus capsid protein" /protein_id="" /transl_table=1 /translation="EGRGSLLTCGDVEENPGP" /label="T2A" CDS 5282..5881 /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 5882..6469 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6543..6776 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6848..6969 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6988..7123 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(7161..7177) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7185..7201 /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(7209..7239) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7254..7275 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7563..8151) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8322..9182) /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(9183..9287) /gene="bla" /label="AmpR promoter" primer_bind 9761..9777 /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 attacaaaaa ttcaaaattt 1921 tatcgatact agtattatgc ccagtacatg accttatggg actttcctac ttggcagtac 1981 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg 2041 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg 2101 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca 2161 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggttt atataagcag agctcgttta 2221 gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagattc 2281 tagagctagc gaattcgcca ccatgagtct gctaaactgt gaaaacagct gtggatccag 2341 ccagtctgaa agtgactgct gtgtggccat ggccagctcc tgtagcgctg taacaaaaga 2401 tgatagtgtg ggtggaactg ccagcacggg gaacctctcc agctcattta tggaggagat 2461 ccagggatat gatgtagagt ttgacccacc cctggaaagc aagtatgaat gccccatctg 2521 cttgatggca ttacgagaag cagtgcaaac gccatgcggc cataggttct gcaaagcctg 2581 catcataaaa tcaataaggg atgcaggtca caaatgtcca gttgacaatg aaatactgct 2641 ggaaaatcaa ctatttccag acaattttgc aaaacgtgag attctttctc tgatggtgaa 2701 atgtccaaat gaaggttgtt tgcacaagat ggaactgaga catcttgagg atcatcaagc 2761 acattgtgag tttgctctta tggattgtcc ccaatgccag cgtcccttcc aaaaattcca 2821 tattaatatt cacattctga aggattgtcc aaggagacag gtttcttgtg acaactgtgc 2881 tgcatcaatg gcatttgaag ataaagagat ccatgaccag aactgtcctt tggcaaatgt 2941 catctgtgaa tactgcaata ctatactcat cagagaacag atgcctaatc attatgatct 3001 agactgccct acagccccaa ttccatgcac attcagtact tttggttgcc atgaaaagat 3061 gcagaggaat cacttggcac gccacctaca agagaacacc cagtcacaca tgagaatgtt 3121 ggcccaggct gttcatagtt tgagcgttat acccgactct gggtatatct cagaggtccg 3181 gaatttccag gaaactattc accagttaga gggtcgcctt gtaagacaag accatcaaat 3241 ccgggagctg actgctaaaa tggaaactca gagtatgtat gtaagtgagc tcaaacgaac 3301 cattcgaacc cttgaggaca aagttgctga aatcgaagca cagcagtgca atggaattta 3361 tatttggaag attggcaact ttggaatgca tttgaaatgt caagaagagg agaaacctgt 3421 tgtgattcat agccctggat tctacactgg caaacccggg tacaaactgt gcatgcgctt 3481 gcaccttcag ttaccgactg ctcagcgctg tgcaaactat atatcccttt ttgtccacac 3541 aatgcaagga gaatatgaca gccacctccc ttggcccttc cagggtacaa tacgccttac 3601 aattcttgat cagtctgaag cacctgtaag gcaaaaccac gaagagataa tggatgccaa 3661 accagagctg cttgctttcc agcgacccac aatcccacgg aacccaaaag gttttggcta 3721 tgtaactttt atgcatctgg aagccctaag acaaagaact ttcattaagg atgacacatt 3781 attagtgcgc tgtgaggtct ccacccgctt tgacatgggt agccttcgga gggagggttt 3841 tcagccacga agtactgatg caggggtagg atccgactac aaagaccatg acggtgatta 3901 taaagatcat gacatcgact acaaggatga cgatgacaag tagcggccgc gaaggatctg 3961 cgatcgctcc ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt 4021 ggggggaggg gtcggcaatt gaacgggtgc ctagagaagg tggcgcgggg taaactggga 4081 aagtgatgtc gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag 4141 tgcagtagtc gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacagctgaa 4201 gcttcgaggg gctcgcatct ctccttcacg cgcccgccgc cctacctgag gccgccatcc 4261 acgccggttg agtcgcgttc tgccgcctcc cgcctgtggt gcctcctgaa ctgcgtccgc 4321 cgtctaggta agtttaaagc tcaggtcgag accgggcctt tgtccggcgc tcccttggag 4381 cctacctaga ctcagccggc tctccacgct ttgcctgacc ctgcttgctc aactctacgt 4441 ctttgtttcg ttttctgttc tgcgccgtta cagatccaag ctgtgaccgg cgcctacgct 4501 agacgccacc atggtgagca agggcgagga ggataacatg gccatcatca aggagttcat 4561 gcgcttcaag gtgcacatgg agggctccgt gaacggccac gagttcgaga tcgagggcga 4621 gggcgagggc cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggtgg 4681 ccccctgccc ttcgcctggg acatcctgtc ccctcagttc atgtacggct ccaaggccta 4741 cgtgaagcac cccgccgaca tccccgacta cttgaagctg tccttccccg agggcttcaa 4801 gtgggagcgc gtgatgaact tcgaggacgg cggcgtggtg accgtgaccc aggactcctc 4861 cctgcaggac ggcgagttca tctacaaggt gaagctgcgc ggcaccaact tcccctccga 4921 cggccccgta atgcagaaga agaccatggg ctgggaggcc tcctccgagc ggatgtaccc 4981 cgaggacggc gccctgaagg gcgagatcaa gcagaggctg aagctgaagg acggcggcca 5041 ctacgacgct gaggtcaaga ccacctacaa ggccaagaag cccgtgcagc tgcccggcgc 5101 ctacaacgtc aacatcaagt tggacatcac ctcccacaac gaggactaca ccatcgtgga 5161 acagtacgaa cgcgccgagg gccgccactc caccggcggc atggacgagc tgtacaaggg 5221 atctcgcgag ggcagaggaa gtcttctaac atgcggtgac gtggaggaga atcccggccc 5281 tatgaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt 5341 acgcaccctc gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgatccgga 5401 ccgccacatc gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga 5461 catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga 5521 gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg 5581 ttcccggctg gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga 5641 gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg 5701 cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct 5761 ggagacctcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc 5821 cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg 5881 aaatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc 5941 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg 6001 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt 6061 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac 6121 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc 6181 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct 6241 gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct 6301 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct 6361 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct 6421 ccgccttcgc cctcagacga gtcggatctc cctttggccg cctccccgcc tggtaccttt 6481 aagaccaatg acttacaagg cagctgtaga tcttagccac tttttaaaag aaaagggggg 6541 actggaaggg ctaattcact cccaacgaaa ataagatctg ctttttgctt gtactgggtc 6601 tctctggtta gaccagatct gagcctggga gctctctggc taactaggga acccactgct 6661 taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga 6721 ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtag 6781 tagttcatgt catcttatta ttcagtattt ataacttgca aagaaatgaa tatcagagag 6841 tgagaggaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa 6901 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa 6961 tgtatcttat catgtctggc tctagctatc ccgcccctaa ctccgcccag ttccgcccat 7021 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc 7081 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctagactt ttgcagagac 7141 ggcccaaatt cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca 7201 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg 7261 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg 7321 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc 7381 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta 7441 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag 7501 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg 7561 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg 7621 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg 7681 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga 7741 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc 7801 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt 7861 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact 7921 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg 7981 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt 8041 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt 8101 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct 8161 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg 8221 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt 8281 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt 8341 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc 8401 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg 8461 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc 8521 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg 8581 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca 8641 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga 8701 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct 8761 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg 8821 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca 8881 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata 8941 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct 9001 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact 9061 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa 9121 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc 9181 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga 9241 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga 9301 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg 9361 cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac 9421 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc 9481 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca 9541 gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg 9601 agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga 9661 tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga 9721 ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc 9781 aagctg //