LOCUS V020282 9844 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V020282 VERSION V020282 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 1798..1915 /note="central polypurine tract and central termination sequence of HIV-1" /label="cPPT/CTS" promoter 2005..2208 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 2289..4031 /codon_start=1 /product="human PTEN-induced putative kinase 1, a mitochondria serine/threonine protein kinase" /transl_table=1 /translation="MAVRQALGRGLQLGRALLLRFTGKPGRAYGLGRPGPAAGCVRGER PGWAAGPGAEPRRVGLGLPNRLRFFRQSVAGLAARLQRQFVVRAWGCAGPCGRAVFLAF GLGLGLIEEKQAESRRAVSACQEIQAIFTQKSKPGPDPLDTRRLQGFRLEEYLIGQSIG KGCSAAVYEATMPTLPQNLEVTKSTGLLPGRGPGTSAPGEGQERAPGAPAFPLAIKMMW NISAGSSSEAILNTMSQELVPASRVALAGEYGAVTYRKSKRGPKQLAPHPNIIRVLRAF TSSVPLLPGALVDYPDVLPSRLHPEGLGHGRTLFLVMKNYPCTLRQYLCVNTPSPRLAA MMLLQLLEGVDHLVQQGIAHRDLKSDNILVELDPDGCPWLVIADFGCCLADESIGLQLP FSSWYVDRGGNGCLMAPEVSTARPGPRAVIDYSKADAWAVGAIAYEIFGLVNPFYGQGK AHLESRSYQEAQLPALPESVPPDVRQLVRALLQREASKRPSARVAANVLHLSLWGEHIL ALKNLKLDKMVGWLLQQSAATLLANRLTEKCCVETKMKMLFLANLECETLCQAALLLCS WRAAL" /label="PINK1" CDS 4047..4766 /codon_start=1 /note="mammalian codon-optimized" /product="enhanced YFP" /transl_table=1 /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL TLKFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFK DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMV LLEFVTAAGITLGMDELYK*" /label="EYFP" promoter 4814..5025 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 5038..5306 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 5333..5932 /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 5939..6527 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6601..6834 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6906..7027 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 7046..7181 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(7219..7235) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7243..7259 /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(7267..7297) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7312..7333 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7621..8209) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8380..9240) /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(9241..9345) /gene="bla" /label="AmpR promoter" primer_bind 9819..9835 /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 ggttaacttt 1801 taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc 1861 aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa attttatcga 1921 tactagtatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac 1981 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga 2041 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg 2101 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg 2161 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctcg tttagtgaac 2221 cgtcagatcg cctggagacg ccatccacgc tgttttgacc tccatagaag attctagagc 2281 tagccaccat ggcggtgcga caggcgctgg gccgcggcct gcagctgggt cgagcgctgc 2341 tgctgcgctt cacgggcaag cccggccggg cctacggctt ggggcggccg ggcccggcgg 2401 cgggctgtgt ccgcggggag cgtccaggct gggccgcagg accgggcgcg gagcctcgca 2461 gggtcgggct cgggctccct aaccgtctcc gcttcttccg ccagtcggtg gccgggctgg 2521 cggcgcggtt gcagcggcag ttcgtggtgc gggcctgggg ctgcgcgggc ccttgcggcc 2581 gggcagtctt tctggccttc gggctagggc tgggcctcat cgaggaaaaa caggcggaga 2641 gccggcgggc ggtctcggcc tgtcaggaga tccaggcaat ttttacccag aaaagcaagc 2701 cggggcctga cccgttggac acgagacgct tgcagggctt tcggctggag gagtatctga 2761 tagggcagtc cattggtaag ggctgcagtg ctgctgtgta tgaagccacc atgcctacat 2821 tgccccagaa cctggaggtg acaaagagca ccgggttgct tccagggaga ggcccaggta 2881 ccagtgcacc aggagaaggg caggagcgag ctccgggggc ccctgccttc cccttggcca 2941 tcaagatgat gtggaacatc tcggcaggtt cctccagcga agccatcttg aacacaatga 3001 gccaggagct ggtcccagcg agccgagtgg ccttggctgg ggagtatgga gcagtcactt 3061 acagaaaatc caagagaggt cccaagcaac tagcccctca ccccaacatc atccgggttc 3121 tccgcgcctt cacctcttcc gtgccgctgc tgccaggggc cctggtcgac taccctgatg 3181 tgctgccctc acgcctccac cctgaaggcc tgggccatgg ccggacgctg ttcctcgtta 3241 tgaagaacta tccctgtacc ctgcgccagt acctttgtgt gaacacaccc agcccccgcc 3301 tcgccgccat gatgctgctg cagctgctgg aaggcgtgga ccatctggtt caacagggca 3361 tcgcgcacag agacctgaaa tccgacaaca tccttgtgga gctggaccca gacggctgcc 3421 cctggctggt gatcgcagat tttggctgct gcctggctga tgagagcatc ggcctgcagt 3481 tgcccttcag cagctggtac gtggatcggg gcggaaacgg ctgtctgatg gccccagagg 3541 tgtccacggc ccgtcctggc cccagggcag tgattgacta cagcaaggct gatgcctggg 3601 cagtgggagc catcgcctat gaaatcttcg ggcttgtcaa tcccttctac ggccagggca 3661 aggcccacct tgaaagccgc agctaccaag aggctcagct acctgcactg cccgagtcag 3721 tgcctccaga cgtgagacag ttggtgaggg cactgctcca gcgagaggcc agcaagagac 3781 catctgcccg agtagccgca aatgtgcttc atctaagcct ctggggtgaa catattctag 3841 ccctgaagaa tctgaagtta gacaagatgg ttggctggct cctccaacaa tcggccgcca 3901 ctttgttggc caacaggctc acagagaagt gttgtgtgga aacaaaaatg aagatgctct 3961 ttctggctaa cctggagtgt gaaacgctct gccaggcagc cctcctcctc tgctcatgga 4021 gggcagccct gggaggaggg ggaagcatgg tgagcaaggg cgaggagctg ttcaccgggg 4081 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg 4141 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg 4201 gcaagctgcc cgtgccctgg cccaccctcg tgaccacctt cggctacggc ctgcagtgct 4261 tcgcccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag 4321 gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg 4381 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca 4441 aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct 4501 atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca 4561 tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg 4621 gccccgtgct gctgcccgac aaccactacc tgagctacca gtccgccctg agcaaagacc 4681 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc 4741 tcggcatgga cgagctgtac aagtaaggat ccgcggccgc gaaggatctg cgatcgctcc 4801 ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg 4861 gtcggcaatt gaacgggtgc ctagagaagg tggcgcgggg taaactggga aagtgatgtc 4921 gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag tgcagtagtc 4981 gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacagctgaa gcttcgaggg 5041 gctcgcatct ctccttcacg cgcccgccgc cctacctgag gccgccatcc acgccggttg 5101 agtcgcgttc tgccgcctcc cgcctgtggt gcctcctgaa ctgcgtccgc cgtctaggta 5161 agtttaaagc tcaggtcgag accgggcctt tgtccggcgc tcccttggag cctacctaga 5221 ctcagccggc tctccacgct ttgcctgacc ctgcttgctc aactctacgt ctttgtttcg 5281 ttttctgttc tgcgccgtta cagatccaag ctgtgaccgg cgcctacgct agatgaccga 5341 gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc ccccgggccg tacgcaccct 5401 cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgacccgg accgccacat 5461 cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa 5521 ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga 5581 agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct 5641 ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg 5701 gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt 5761 cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc 5821 cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga 5881 ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gagtcgacaa 5941 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc 6001 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat 6061 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg 6121 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg 6181 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat 6241 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt 6301 gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc 6361 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa 6421 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg 6481 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg gtacctttaa 6541 gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac 6601 tggaagggct aattcactcc caacgaaaat aagatctgct ttttgcttgt actgggtctc 6661 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta 6721 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact 6781 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtagta 6841 gttcatgtca tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg 6901 agaggaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt 6961 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg 7021 tatcttatca tgtctggctc tagctatccc gcccctaact ccgcccagtt ccgcccattc 7081 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc 7141 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctagactttt gcagagacgg 7201 cccaaattcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat 7261 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag 7321 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg 7381 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc 7441 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc 7501 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa 7561 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt 7621 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg 7681 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg 7741 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag 7801 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc 7861 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa 7921 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg 7981 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc 8041 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac 8101 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg 8161 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt 8221 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt 8281 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa 8341 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga 8401 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt 8461 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg 8521 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga 8581 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga 8641 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg 8701 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc 8761 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc 8821 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca 8881 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac 8941 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg 9001 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc 9061 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg 9121 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac 9181 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat 9241 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata 9301 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa 9361 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg 9421 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat 9481 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg 9541 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga 9601 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag 9661 aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc 9721 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt 9781 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa 9841 gctg //