LOCUS V026714 9778 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V026714 VERSION V026714 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 2317..3804 /label="Xiap(NM_022231)" /note="Xiap(NM_022231)" /gene="Xiap" CDS 3811..3876 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3920..4131 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4144..4412 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4441..4450 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4471..5133 /codon_start=1 /product="green fluorescent protein 2 from Pontellina plumata, also known as ppluGFP2 (Shagin et al., 2004)" /transl_table=1 /translation="PAMEIECRITGTLNGVEFELVGGGEGTPKQGRMTNKMKSTKGALT FSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVLHVSFSYRY EAGRVIGDFKVVGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNVLVGSFARTFSLRDGG YYSFVVDSHMHFKSAIHPSILQNGGPMFAFRRVEELHSNTELGIVEYQHAFKTPIAFA" /label="CopGFP" CDS 5203..5256 /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 5257..5856 /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 5857..6445 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6519..6752 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6824..6945 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6985..7120 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(7153..7169) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7177..7193 /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(7201..7231) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7246..7267 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7555..8143) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8314..9174) /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(9175..9279) /gene="bla" /label="AmpR promoter" primer_bind 9753..9769 /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 ctgatacgaa ctcggaattc gccaccatga cttttaacag ttttgaagga 2341 tctagaactg ttgtacctgc agacaccaat aaggatgaag aatttgtaga agagtttaat 2401 agattaaaaa catttgctaa cttcccaagc agcagtcctg tttcagcatc aacattggcg 2461 cgagcggggt ttctctacac tggtgaagga gacaccgtgc agtgtttcag ttgtcacgcg 2521 gcagtagata gatggcagta tggagactca gctgttggaa gacacaggag aatatcccca 2581 aattgcagat ttatcaatgg tttttatttt gaaaacggtg ccacacagtc tacatctcct 2641 ggcatccaaa atggccagta caaatctgaa aactgtgtgg gaaacagaaa tcattttgct 2701 cttgacaggc cgtcggagac tcatgcagat tatctcctga gaactggaca ggttgtagat 2761 atttcagata ccatataccc gaggaacccg gccatgtgta gtgaagaagc cagactgaag 2821 acgtttcaga actggccaga ctatgcccat ttaagcccca gagagttagc tagtgctgga 2881 ctctactaca cggggattga tgatcaagtg caatgctttt gttgtggtgg aaaactgaaa 2941 aattgggaac cctgtgaccg tgcctggtca gagcacagga gacactttcc caactgcttc 3001 ttcgttttgg gccggaatgt taatgttcga agtgagtctg gtgtgagttc agataggaat 3061 ttcccaaatt caacaaattc tccaagaaat ccagccatgg cagaatatga cgcacggatc 3121 gttacttttg gaacatggct atactcagtt aacaaggagc agcttgcaag agctggattt 3181 tatgctttag gtgaaggtga taaagtgaag tgctttcact gtggaggagg gctcacggat 3241 tggaagccaa gtgaagaccc ttgggaacag catgctaagt ggtatccagg gtgtaaatat 3301 ctattggatg agaagggaca agaatatata aataatattc atttaaccca ttcacttggg 3361 gaatctgtgg taagaactgc tgaaaaaaca ccatcagtaa ctaaaaaaat cgatgatacc 3421 atcttccaga atcctatggt gcaagaagct atacgaatgg gattcaactt caaggacatc 3481 aagaaaacaa tggaagaaaa gctccaaaca tctgggagca actatctatc acttgaggtt 3541 ctgattgcag atcttgtgag tgctcagaaa gataattcgc aggatgagtc aagtcagact 3601 tcattgcaga aagacatcag tactgaagag cagctaaggc gcctacaaga ggagaagctt 3661 tgcaaaatct gtatggatag aaatattgct atagtttttg ttccttgtgg acatctggtc 3721 acttgtaaac agtgtgcgga agcagttgac aaatgtccca tgtgctgcac agtcattacg 3781 ttcaagcaaa aaatttttat gtctggatcc gactacaaag accatgacgg tgattataaa 3841 gatcatgaca tcgactacaa ggatgacgat gacaagtagc ggccgcgaag gatctgcgat 3901 cgctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg 3961 ggaggggtcg gcaattgaac gggtgcctag agaaggtggc gcggggtaaa ctgggaaagt 4021 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca 4081 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca gctgaagctt 4141 cgaggggctc gcatctctcc ttcacgcgcc cgccgcccta cctgaggccg ccatccacgc 4201 cggttgagtc gcgttctgcc gcctcccgcc tgtggtgcct cctgaactgc gtccgccgtc 4261 taggtaagtt taaagctcag gtcgagaccg ggcctttgtc cggcgctccc ttggagccta 4321 cctagactca gccggctctc cacgctttgc ctgaccctgc ttgctcaact ctacgtcttt 4381 gtttcgtttt ctgttctgcg ccgttacaga tccaagctgt gaccggcgcc tacgctagac 4441 gccaccatgg agagcgacga gagcggcctg cccgccatgg agatcgagtg ccgcatcacc 4501 ggcaccctga acggcgtgga gttcgagctg gtgggcggcg gagagggcac ccccaagcag 4561 ggccgcatga ccaacaagat gaagagcacc aaaggcgccc tgaccttcag cccctacctg 4621 ctgagccacg tgatgggcta cggcttctac cacttcggca cctaccccag cggctacgag 4681 aaccccttcc tgcacgccat caacaacggc ggctacacca acacccgcat cgagaagtac 4741 gaggacggcg gcgtgctgca cgtgagcttc agctaccgct acgaggccgg ccgcgtgatc 4801 ggcgacttca aggtggtggg caccggcttc cccgaggaca gcgtgatctt caccgacaag 4861 atcatccgca gcaacgccac cgtggagcac ctgcacccca tgggcgataa cgtgctggtg 4921 ggcagcttcg cccgcacctt cagcctgcgc gacggcggct actacagctt cgtggtggac 4981 agccacatgc acttcaagag cgccatccac cccagcatcc tgcagaacgg gggccccatg 5041 ttcgccttcc gccgcgtgga ggagctgcac agcaacaccg agctgggcat cgtggagtac 5101 cagcacgcct tcaagacccc catcgccttc gccagatccc gcgctcagtc gtccaattct 5161 gccgtggacg gcaccgccgg acccggctcc accggatctc gcgagggcag aggaagtctt 5221 ctaacatgcg gtgacgtgga ggagaatccc ggccctatga ccgagtacaa gcccacggtg 5281 cgcctcgcca cccgcgacga cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc 5341 gactaccccg ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag 5401 ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac 5461 gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc 5521 gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc gcagcaacag 5581 atggaaggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct ggccaccgtc 5641 ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg 5701 gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc 5761 cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg 5821 cgcacctggt gcatgacccg caagcccggt gcctgaaatc aacctctgga ttacaaaatt 5881 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct 5941 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg 6001 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc 6061 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt 6121 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc 6181 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg 6241 ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac ctggattctg 6301 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc 6361 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg 6421 atctcccttt gggccgcctc cccgcctggt acctttaaga ccaatgactt acaaggcagc 6481 tgtagatctt agccactttt taaaagaaaa ggggggactg gaagggctaa ttcactccca 6541 acgaagataa gatctgcttt ttgcttgtac tgggtctctc tggttagacc agatctgagc 6601 ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg 6661 agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag 6721 acccttttag tcagtgtgga aaatctctag cagtagtagt tcatgtcatc ttattattca 6781 gtatttataa cttgcaaaga aatgaatatc agagagtgag aggaacttgt ttattgcagc 6841 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc 6901 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggctcta 6961 gctatcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct 7021 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct 7081 gagctattcc agaagtagtg aggaggcttt tttggaggcc tagacttttg cagagaccaa 7141 attcgtaatc atgtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca 7201 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact 7261 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct 7321 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc 7381 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca 7441 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg 7501 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca 7561 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa 7621 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc 7681 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc 7741 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct 7801 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg 7861 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag 7921 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta 7981 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg 8041 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt 8101 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt 8161 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag 8221 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat 8281 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc 8341 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat 8401 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc 8461 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag 8521 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag 8581 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt 8641 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg 8701 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt 8761 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc 8821 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc 8881 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa 8941 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg 9001 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc 9061 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag 9121 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt 9181 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt 9241 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc 9301 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac 9361 gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct 9421 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg 9481 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat 9541 tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata 9601 ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg 9661 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg 9721 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg ccaagctg //