LOCUS V038546 9954 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V038546 VERSION V038546 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 2297..3973 /label="YTHDF1(NM_017798)" /note="YTHDF1(NM_017798)" /gene="YTHDF1" CDS 3980..4069 /codon_start=1 /product="3 tandem Myc epitope tags" /transl_table=1 /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL" /label="3xMyc" promoter 4113..4324 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4337..4605 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4634..4643 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4664..5326 /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 5396..5449 /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 5450..6049 /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 6050..6637 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6711..6944 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 7016..7137 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 7156..7291 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(7329..7345) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7353..7369 /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(7377..7407) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7422..7443 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7731..8319) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8490..9350) /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(9351..9455) /gene="bla" /label="AmpR promoter" primer_bind 9929..9945 /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 tagcgaattc gccaccatgt cggccaccag cgtggacacc cagagaacaa aaggacaaga 2341 taataaagta caaaatggtt cgttacatca gaaggataca gttcatgaca atgactttga 2401 gccctacctt actggacagt caaatcagag taacagttac ccctcaatga gcgaccccta 2461 cctgtccagc tattacccgc cgtccattgg atttccttac tccctcaatg aggctccgtg 2521 gtctactgca ggggaccctc cgattccata cctcaccacc tacggacagc tcagtaacgg 2581 agaccatcat tttatgcacg atgctgtttt tgggcagcct gggggcctgg ggaacaacat 2641 ctatcagcac aggttcaatt ttttccctga aaaccctgcg ttctcagcat gggggacaag 2701 tgggtctcaa ggtcagcaga cccagagctc cgcgtatggg agcagctaca cctacccccc 2761 gagctccctg ggtggcacgg tggttgatgg gcagccaggc tttcacagcg acaccctcag 2821 caaggccccc gggatgaaca gcctggagca gggcatggtt ggcctgaaga ttggggacgt 2881 cagctcctcc gccgtcaaga cggtgggctc tgtcgtcagc agcgtggcac tgactggtgt 2941 cctttctggc aacggtggga caaatgtgaa catgccagtt tcaaagccga cctcgtgggc 3001 tgccattgcc agcaagcctg caaaaccaca gcctaaaatg aaaacaaaga gcgggcctgt 3061 catggggggt gggctgcccc ctccacccat aaagcataac atggacattg gcacctggga 3121 taacaagggg cctgtgccga aggccccagt cccccagcag gcaccctctc cacaggctgc 3181 cccacagccc cagcaggtgg ctcagcctct cccagcacag cccccagctt tggctcaacc 3241 gcagtatcag agccctcagc agccacccca gacccgctgg gttgccccac gcaacagaaa 3301 cgcggcgttt gggcagagcg gaggggctgg cagcgatagc aactctcctg gaaacgtcca 3361 gcctaattct gcccccagcg tcgaatccca ccccgtcctt gaaaaactga aggctgctca 3421 cagctacaac ccgaaagagt ttgagtggaa tctgaaaagc gggcgtgtgt tcatcatcaa 3481 gagctactct gaggacgaca tccaccgctc cattaagtac tccatctggt gtagcacaga 3541 gcacggcaac aagcgcctgg acagcgcctt ccgctgcatg agcagcaagg ggcccgtcta 3601 cctgctcttc agcgtcaatg ggagtgggca tttttgtggg gtggccgaga tgaagtcccc 3661 cgtggactac ggcaccagtg ccggggtctg gtctcaggac aagtggaagg ggaagtttga 3721 tgtccagtgg atttttgtta aggatgtacc caataaccag ctccggcaca tcaggctgga 3781 gaataacgac aacaaaccgg tcacaaactc ccgggacacc caggaggtgc ccttagaaaa 3841 agccaagcaa gtgctgaaaa ttatcagttc ctacaagcac acaacctcca tcttcgacga 3901 ctttgctcac tacgagaagc gccaggagga ggaggaggtg gtgcgcaagg aacggcagag 3961 tcgaaacaaa caaggatccg aacaaaaact aatatcagag gaagatttgg aacagaaact 4021 aatcagtgaa gaagatttag agcaaaagtt aatttccgag gaggacttat agcggccgcg 4081 aaggatctgc gatcgctccg gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc 4141 cgagaagttg gggggagggg tcggcaattg aacgggtgcc tagagaaggt ggcgcggggt 4201 aaactgggaa agtgatgtcg tgtactggct ccgccttttt cccgagggtg ggggagaacc 4261 gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg ccgccagaac 4321 acagctgaag cttcgagggg ctcgcatctc tccttcacgc gcccgccgcc ctacctgagg 4381 ccgccatcca cgccggttga gtcgcgttct gccgcctccc gcctgtggtg cctcctgaac 4441 tgcgtccgcc gtctaggtaa gtttaaagct caggtcgaga ccgggccttt gtccggcgct 4501 cccttggagc ctacctagac tcagccggct ctccacgctt tgcctgaccc tgcttgctca 4561 actctacgtc tttgtttcgt tttctgttct gcgccgttac agatccaagc tgtgaccggc 4621 gcctacgcta gacgccacca tggagagcga cgagagcggc ctgcccgcca tggagatcga 4681 gtgccgcatc accggcaccc tgaacggcgt ggagttcgag ctggtgggcg gcggagaggg 4741 cacccccaag cagggccgca tgaccaacaa gatgaagagc accaaaggcg ccctgacctt 4801 cagcccctac ctgctgagcc acgtgatggg ctacggcttc taccacttcg gcacctaccc 4861 cagcggctac gagaacccct tcctgcacgc catcaacaac ggcggctaca ccaacacccg 4921 catcgagaag tacgaggacg gcggcgtgct gcacgtgagc ttcagctacc gctacgaggc 4981 cggccgcgtg atcggcgact tcaaggtggt gggcaccggc ttccccgagg acagcgtgat 5041 cttcaccgac aagatcatcc gcagcaacgc caccgtggag cacctgcacc ccatgggcga 5101 taacgtgctg gtgggcagct tcgcccgcac cttcagcctg cgcgacggcg gctactacag 5161 cttcgtggtg gacagccaca tgcacttcaa gagcgccatc caccccagca tcctgcagaa 5221 cgggggcccc atgttcgcct tccgccgcgt ggaggagctg cacagcaaca ccgagctggg 5281 catcgtggag taccagcacg ccttcaagac ccccatcgcc ttcgccagat cccgcgctca 5341 gtcgtccaat tctgccgtgg acggcaccgc cggacccggc tccaccggat ctcgcgaggg 5401 cagaggaagt cttctaacat gcggtgacgt ggaggagaat cccggcccta tgaccgagta 5461 caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac gcaccctcgc 5521 cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gatccggacc gccacatcga 5581 gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt 5641 gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc 5701 gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc 5761 cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt 5821 cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt 5881 gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc 5941 gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt 6001 gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgaa atcaacctct 6061 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct 6121 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat 6181 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt 6241 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat 6301 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc 6361 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga 6421 caattccgtg gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc 6481 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga 6541 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtctcc gccttcgccc 6601 tcagacgagt cggatctccc tttggccgcc tccccgcctg gtacctttaa gaccaatgac 6661 ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct 6721 aattcactcc caacgaaaat aagatctgct ttttgcttgt actgggtctc tctggttaga 6781 ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata 6841 aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta 6901 gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtagta gttcatgtca 6961 tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggaactt 7021 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa 7081 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca 7141 tgtctggctc tagctatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat 7201 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc 7261 cagaagtagt gaggaggctt ttttggaggc ctagactttt gcagagacgg cccaaattcg 7321 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac 7381 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca 7441 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat 7501 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc 7561 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca 7621 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca 7681 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg 7741 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg 7801 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt 7861 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt 7921 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc 7981 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt 8041 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt 8101 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc 8161 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa 8221 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt 8281 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct 8341 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta 8401 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa 8461 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc 8521 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact 8581 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc 8641 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt 8701 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta 8761 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg 8821 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt 8881 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc 8941 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt 9001 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc 9061 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc 9121 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa 9181 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac 9241 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa 9301 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt 9361 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa 9421 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct 9481 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg 9541 ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg 9601 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg 9661 tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta 9721 ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc 9781 atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc 9841 tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta 9901 acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa gctg //