LOCUS V023888 8829 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V023888 VERSION V023888 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 2301..3683 /label="Pik3r3(NM_001355585)" /note="Pik3r3(NM_001355585)" /gene="Pik3r3" CDS 3690..3755 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3799..4010 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4023..4291 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 4318..4917 /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 4924..5512 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5586..5819 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 5891..6012 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6031..6166 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6204..6220) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6228..6244 /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(6252..6282) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6297..6318 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6606..7194) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7365..8225) /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(8226..8330) /gene="bla" /label="AmpR promoter" primer_bind 8804..8820 /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 tgaccgtcag atcgcctgga gacgccatcc acgctgtttt gacctccata gaagattcta 2281 gagctagcga attcgccacc atgtacaata cggtgtggag tatggaccgc gatgacgcag 2341 actggaggga ggtgatgatg ccctattcga cagaactgat attttatatt gaaatggatc 2401 ctccagctct tccaccaaag ccacctaagc caatgactcc agcagtcaca aatggaatga 2461 aggacagttt catttctctt caagatgcag agtggtactg gggagatatt tccagggaag 2521 aggtaaatga caaattgcgg gacatgccag atggtacctt cttagttcgt gatgcctcaa 2581 cgaaaatgca gggggattat acattgactt tgaggaaggg aggaaataat aaattaataa 2641 agatctatca tcgggatggt aaatatggct tctctgagcc cctgacgttt acttctgtgg 2701 tggagcttat taaccactac caccacgagt ctctcgctca gtacaatccc aaactcgacg 2761 tgaagctgac gtacccagta tccagattcc aacaggatca gttggtaaaa gaagataaca 2821 ttgatgcagt aggtaaaaat ctgcaggagt tccactctca gtatcaggag aagagcaaag 2881 agtatgacag gctgtatgaa gagtacacaa ggacatcaca ggaaatacaa atgaagagga 2941 ctgccattga agcctttaat gaaacaatta aaatatttga ggagcagtgt catacccaag 3001 aacaacacag taaagactat atcgagcgct ttcgcagaga ggggaatgag aaggagatcg 3061 agcgaattat gatgaattat gataaattga aatcacgtct tggtgagatt catgatagca 3121 aactgcgtct tgagcaggac ttgaagaaac aagctttgga caaccgggaa atagataaaa 3181 aaatgaatag catcaaaccc gacttgatcc agctgcgtaa gatccgggat cagcaccttg 3241 tatggctcaa tcacagagga gtgaggcaga ggcgcctgaa tgcctggctg gggatcaaga 3301 atgaggactc agatgaaagc tattttatca atgaggaaga tgagaacctg ccgcattatg 3361 atgagaaaac ctggtttgtg gaggatatca accgagtaca agcagaggac ttgctttatg 3421 ggaaaccaga tggtgcattc ttaattcgtg agagtagcaa gaaaggatgt tacgcttgtt 3481 ctgtggttgc agacggggaa gtgaagcact gtgtcatcta cagcacggct cgaggatatg 3541 gctttgcaga accctacaac ctgtacagct cactgaagga gctggtgctc cattaccagc 3601 agacatccct ggttcagcac aacgactccc tcaacgtcag gctcgcctac cctgtccatg 3661 cacagatgcc tacgctctgc agaggatccg actacaaaga ccatgacggt gattataaag 3721 atcatgacat cgactacaag gatgacgatg acaagtagcg gccgcgaagg atctgcgatc 3781 gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag aagttggggg 3841 gaggggtcgg caattgaacg ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg 3901 atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat ataagtgcag 3961 tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag ctgaagcttc 4021 gaggggctcg catctctcct tcacgcgccc gccgccctac ctgaggccgc catccacgcc 4081 ggttgagtcg cgttctgccg cctcccgcct gtggtgcctc ctgaactgcg tccgccgtct 4141 aggtaagttt aaagctcagg tcgagaccgg gcctttgtcc ggcgctccct tggagcctac 4201 ctagactcag ccggctctcc acgctttgcc tgaccctgct tgctcaactc tacgtctttg 4261 tttcgttttc tgttctgcgc cgttacagat ccaagctgtg accggcgcct acgctagatg 4321 accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtcccccg ggccgtacgc 4381 accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga cccggaccgc 4441 cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc 4501 ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc 4561 gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc 4621 cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc caaggagccc 4681 gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc agggcaaggg tctgggcagc 4741 gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag 4801 acctccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac 4861 gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg tgcctgagtc 4921 gacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt 4981 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc 5041 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag 5101 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc 5161 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc 5221 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg 5281 ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg 5341 ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc 5401 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt 5461 cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc gcctggtacc 5521 tttaagacca atgacttaca aggcagctgt agatcttagc cactttttaa aagaaaaggg 5581 gggactggaa gggctaattc actcccaacg aaaataagat ctgctttttg cttgtactgg 5641 gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact 5701 gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg 5761 tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag 5821 tagtagttca tgtcatctta ttattcagta tttataactt gcaaagaaat gaatatcaga 5881 gagtgagagg aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac 5941 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat 6001 caatgtatct tatcatgtct ggctctagct atcccgcccc taactccgcc cagttccgcc 6061 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg 6121 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctaga cttttgcaga 6181 gacggcccaa attcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc 6241 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga 6301 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg 6361 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg 6421 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg 6481 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga 6541 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg 6601 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag 6661 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc 6721 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg 6781 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt 6841 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc 6901 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc 6961 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg 7021 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca 7081 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc 7141 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat 7201 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt 7261 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt 7321 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc 7381 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc 7441 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata 7501 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg 7561 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc 7621 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct 7681 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa 7741 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt 7801 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca 7861 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac 7921 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca 7981 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt 8041 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc 8101 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca 8161 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata 8221 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc 8281 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc 8341 cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat 8401 aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga 8461 cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa 8521 gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca 8581 tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta 8641 aggagaaaat accgcatcag gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg 8701 cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg 8761 cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt 8821 gccaagctg //