LOCUS V027195 9451 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V027195 VERSION V027195 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..3477 /label="DNMT3L(NM_013369)" /note="DNMT3L(NM_013369)" /gene="DNMT3L" CDS 3484..3549 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3593..3804 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3817..4085 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4114..4123 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4144..4806 /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 4876..4929 /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 4930..5529 /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 5530..6118 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6192..6425 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6497..6618 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6658..6793 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6826..6842) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6850..6866 /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(6874..6904) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6919..6940 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7228..7816) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7987..8847) /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(8848..8952) /gene="bla" /label="AmpR promoter" primer_bind 9426..9442 /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 gccaccatgg cggccatccc agccctggac 2341 ccagaggccg agcccagcat ggacgtgatt ttggtgggat ccagtgagct ctcaagctcc 2401 gtttcacccg ggacaggcag agatcttatt gcatatgaag tcaaggctaa ccagcgaaat 2461 atagaagaca tctgcatctg ctgcggaagt ctccaggttc acacacagca ccctctgttt 2521 gagggaggga tctgcgcccc atgtaaggac aagttcctgg atgccctctt cctgtacgac 2581 gatgacgggt accaatccta ctgctccatc tgctgctccg gagagacgct gctcatctgc 2641 ggaaaccctg attgcacccg atgctactgc ttcgagtgtg tggatagcct ggtcggcccc 2701 gggacctcgg ggaaggtgca cgccatgagc aactgggtgt gctacctgtg cctgccgtcc 2761 tcccgaagcg ggctgctgca gcgtcggagg aagtggcgca gccagctcaa ggccttctac 2821 gaccgagagt cggagaatcc ccttgagatg ttcgaaaccg tgcctgtgtg gaggagacag 2881 ccagtccggg tgctgtccct ttttgaagac atcaagaaag agctgacgag tttgggcttt 2941 ttggaaagtg gttctgaccc gggacaactg aagcatgtgg ttgatgtcac agacacagtg 3001 aggaaggatg tggaggagtg gggacccttc gatcttgtgt acggcgccac acctcccctg 3061 ggccacacct gtgaccgtcc tcccagctgg tacctgttcc agttccaccg gctcctgcag 3121 tacgcacggc ccaagccagg cagccccagg cccttcttct ggatgttcgt ggacaatctg 3181 gtgctgaaca aggaagacct ggacgtcgca tctcgcttcc tggagatgga gccagtcacc 3241 atcccagatg tccacggcgg atccttgcag aatgctgtcc gcgtgtggag caacatccca 3301 gccataagga gcagcaggca ctgggctctg gtttcggaag aagaattgtc cctgctggcc 3361 cagaacaagc agagctcgaa gctcgcggcc aagtggccca ccaagctggt gaagaactgc 3421 tttctccccc taagagaata tttcaagtat ttttcaacag aactcacttc ctctttagga 3481 tccgactaca aagaccatga cggtgattat aaagatcatg acatcgacta caaggatgac 3541 gatgacaagt agcggccgcg aaggatctgc gatcgctccg gtgcccgtca gtgggcagag 3601 cgcacatcgc ccacagtccc cgagaagttg gggggagggg tcggcaattg aacgggtgcc 3661 tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg tgtactggct ccgccttttt 3721 cccgagggtg ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc 3781 aacgggtttg ccgccagaac acagctgaag cttcgagggg ctcgcatctc tccttcacgc 3841 gcccgccgcc ctacctgagg ccgccatcca cgccggttga gtcgcgttct gccgcctccc 3901 gcctgtggtg cctcctgaac tgcgtccgcc gtctaggtaa gtttaaagct caggtcgaga 3961 ccgggccttt gtccggcgct cccttggagc ctacctagac tcagccggct ctccacgctt 4021 tgcctgaccc tgcttgctca actctacgtc tttgtttcgt tttctgttct gcgccgttac 4081 agatccaagc tgtgaccggc gcctacgcta gacgccacca tggagagcga cgagagcggc 4141 ctgcccgcca tggagatcga gtgccgcatc accggcaccc tgaacggcgt ggagttcgag 4201 ctggtgggcg gcggagaggg cacccccaag cagggccgca tgaccaacaa gatgaagagc 4261 accaaaggcg ccctgacctt cagcccctac ctgctgagcc acgtgatggg ctacggcttc 4321 taccacttcg gcacctaccc cagcggctac gagaacccct tcctgcacgc catcaacaac 4381 ggcggctaca ccaacacccg catcgagaag tacgaggacg gcggcgtgct gcacgtgagc 4441 ttcagctacc gctacgaggc cggccgcgtg atcggcgact tcaaggtggt gggcaccggc 4501 ttccccgagg acagcgtgat cttcaccgac aagatcatcc gcagcaacgc caccgtggag 4561 cacctgcacc ccatgggcga taacgtgctg gtgggcagct tcgcccgcac cttcagcctg 4621 cgcgacggcg gctactacag cttcgtggtg gacagccaca tgcacttcaa gagcgccatc 4681 caccccagca tcctgcagaa cgggggcccc atgttcgcct tccgccgcgt ggaggagctg 4741 cacagcaaca ccgagctggg catcgtggag taccagcacg ccttcaagac ccccatcgcc 4801 ttcgccagat cccgcgctca gtcgtccaat tctgccgtgg acggcaccgc cggacccggc 4861 tccaccggat ctcgcgaggg cagaggaagt cttctaacat gcggtgacgt ggaggagaat 4921 cccggcccta tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc 4981 agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc 5041 gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc 5101 gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc 5161 acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag 5221 ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg 5281 cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag 5341 ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc 5401 gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc 5461 gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc 5521 ggtgcctgaa atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac 5581 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt 5641 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat 5701 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca 5761 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc 5821 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg 5881 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct 5941 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct 6001 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt 6061 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct 6121 ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga 6181 aaagggggga ctggaagggc taattcactc ccaacgaaga taagatctgc tttttgcttg 6241 tactgggtct ctctggttag accagatctg agcctgggag ctctctggct aactagggaa 6301 cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct 6361 gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc 6421 tagcagtagt agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat 6481 atcagagagt gagaggaact tgtttattgc agcttataat ggttacaaat aaagcaatag 6541 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa 6601 actcatcaat gtatcttatc atgtctggct ctagctatcc cgcccctaac tccgcccatc 6661 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt 6721 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc 6781 ttttttggag gcctagactt ttgcagagac caaattcgta atcatgtcat agctgtttcc 6841 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg 6901 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc 6961 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg 7021 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc 7081 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac 7141 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa 7201 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca 7261 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc 7321 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata 7381 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta 7441 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca 7501 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga 7561 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg 7621 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg 7681 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg 7741 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag 7801 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa 7861 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat 7921 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc 7981 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc 8041 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc 8101 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc 8161 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc 8221 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt 8281 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc 8341 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa 8401 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt 8461 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg 8521 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc 8581 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa 8641 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt 8701 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt 8761 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag 8821 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta 8881 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat 8941 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat 9001 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg 9061 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta 9121 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg 9181 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg 9241 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag 9301 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc 9361 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg 9421 acgttgtaaa acgacggcca gtgccaagct g //