LOCUS V043095 8545 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V043095 VERSION V043095 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" CDS 2302..3399 /label="IDH3A(NM_005530)" /note="IDH3A(NM_005530)" /gene="IDH3A" CDS 3406..3471 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3515..3726 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3739..4007 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 4034..4633 /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 4640..5228 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5302..5535 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 5607..5728 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 5747..5882 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(5920..5936) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5944..5960 /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(5968..5998) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6013..6034 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6322..6910) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7081..7941) /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(7942..8046) /gene="bla" /label="AmpR promoter" primer_bind 8520..8536 /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 catggctggg cccgcgtgga tctctaaggt ctctcggctg 2341 ctgggggcat tccacaaccc aaaacaggtg accagaggtt ttactggtgg tgttcagaca 2401 gtaactttaa ttccaggaga tggtattggc ccagaaattt cagctgcagt tatgaagatt 2461 tttgatgctg ccaaagcacc tattcagtgg gaggagcgga acgtcactgc cattcaagga 2521 cctggaggaa agtggatgat cccttcagag gctaaagagt ccatggataa gaacaagatg 2581 ggcttgaaag gccctttgaa gaccccaata gcagccggtc acccatctat gaatttactg 2641 ctgcgcaaaa catttgacct ttacgcgaat gtccgaccat gtgtctctat cgaaggctat 2701 aaaacccctt acaccgatgt aaatattgtg accattcgag agaacacaga aggagaatac 2761 agtggaattg agcatgtgat tgttgatgga gtcgtgcaga gtatcaagct catcaccgag 2821 ggggcgagca agcgcattgc tgagtttgcc tttgagtatg cccggaacaa ccaccggagc 2881 aacgtcacgg cggtgcacaa agccaacatc atgcggatgt cagatgggct ttttctacaa 2941 aaatgcaggg aagttgcaga aagctgtaaa gatattaaat ttaatgagat gtaccttgat 3001 acagtatgtt tgaatatggt acaagatcct tcccaatttg atgttcttgt tatgccaaat 3061 ttgtatggag acatccttag tgacttgtgt gcaggattga tcggaggtct cggtgtgaca 3121 ccaagtggca acattggagc caatggggtt gcaatttttg agtcggttca tgggacggct 3181 ccagacattg caggcaagga catggcgaat cccacagccc tcctgctcag tgccgtgatg 3241 atgctgcgcc acatgggact ttttgaccat gctgcaagaa ttgaggctgc gtgttttgct 3301 acaattaagg acggaaagag cttgacaaaa gatttgggag gcaatgcaaa atgctcagac 3361 ttcacagagg aaatctgtcg ccgagtaaaa gatttagatg gatccgacta caaagaccat 3421 gacggtgatt ataaagatca tgacatcgac tacaaggatg acgatgacaa gtagcggccg 3481 cgaaggatct gcgatcgctc cggtgcccgt cagtgggcag agcgcacatc gcccacagtc 3541 cccgagaagt tggggggagg ggtcggcaat tgaacgggtg cctagagaag gtggcgcggg 3601 gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa 3661 ccgtatataa gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt tgccgccaga 3721 acacagctga agcttcgagg ggctcgcatc tctccttcac gcgcccgccg ccctacctga 3781 ggccgccatc cacgccggtt gagtcgcgtt ctgccgcctc ccgcctgtgg tgcctcctga 3841 actgcgtccg ccgtctaggt aagtttaaag ctcaggtcga gaccgggcct ttgtccggcg 3901 ctcccttgga gcctacctag actcagccgg ctctccacgc tttgcctgac cctgcttgct 3961 caactctacg tctttgtttc gttttctgtt ctgcgccgtt acagatccaa gctgtgaccg 4021 gcgcctacgc tagatgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt 4081 cccccgggcc gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac 4141 cgtcgacccg gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg 4201 cgtcgggctc gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg 4261 gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc 4321 cgagttgagc ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca 4381 ccggcccaag gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg 4441 caagggtctg ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt 4501 gcccgccttc ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt 4561 caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa 4621 gcccggtgcc tgagtcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg 4681 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta 4741 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct 4801 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt 4861 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac 4921 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg 4981 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc 5041 gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg 5101 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct 5161 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc 5221 ctccccgcct ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact 5281 ttttaaaaga aaagggggga ctggaagggc taattcactc ccaacgaaaa taagatctgc 5341 tttttgcttg tactgggtct ctctggttag accagatctg agcctgggag ctctctggct 5401 aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt 5461 gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt 5521 ggaaaatctc tagcagtagt agttcatgtc atcttattat tcagtattta taacttgcaa 5581 agaaatgaat atcagagagt gagaggaact tgtttattgc agcttataat ggttacaaat 5641 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg 5701 gtttgtccaa actcatcaat gtatcttatc atgtctggct ctagctatcc cgcccctaac 5761 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga 5821 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg 5881 cctagacttt tgcagagacg gcccaaattc gtaatcatgg tcatagctgt ttcctgtgtg 5941 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc 6001 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt 6061 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg 6121 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt 6181 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc 6241 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa 6301 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa 6361 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc 6421 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc 6481 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag 6541 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga 6601 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc 6661 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac 6721 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg 6781 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca 6841 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa 6901 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa 6961 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt 7021 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag 7081 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat 7141 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc 7201 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa 7261 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca 7321 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa 7381 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt 7441 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc 7501 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact 7561 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc 7621 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg 7681 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct 7741 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc 7801 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag 7861 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac 7921 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg 7981 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt 8041 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac 8101 attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga 8161 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga 8221 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg 8281 gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat 8341 accgcacaga tgcgtaagga gaaaataccg catcaggcgc cattcgccat tcaggctgcg 8401 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg 8461 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg 8521 taaaacgacg gccagtgcca agctg //