LOCUS V018244 9453 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V018244 VERSION V018244 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..3538 /label="IDH1(NM_005896)" /note="IDH1(NM_005896)" /gene="IDH1" CDS 3539..3562 /codon_start=1 /product="FLAG® epitope tag, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDDDDK" /label="FLAG" promoter 3612..3823 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3836..4104 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4133..4142 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4163..4825 /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 4895..4948 /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 4949..5548 /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 5549..6136 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6210..6443 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6515..6636 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6655..6790 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6828..6844) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6852..6868 /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(6876..6906) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6921..6942 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7230..7818) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7989..8849) /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(8850..8954) /gene="bla" /label="AmpR promoter" primer_bind 9428..9444 /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 ccaaaaaaat cagtggcggt tctgtggtag agatgcaagg 2341 agatgaaatg acacgaatca tttgggaatt gattaaagag aaactcattt ttccctacgt 2401 ggaattggat ctacatagct atgatttagg catagagaat cgtgatgcca ccaacgacca 2461 agtcaccaag gatgctgcag aagctataaa gaagcataat gttggcgtca aatgtgccac 2521 tatcactcct gatgagaaga gggttgagga gttcaagttg aaacaaatgt ggaaatcacc 2581 aaatggcacc atacgaaata ttctgggtgg cacggtcttc agagaagcca ttatctgcaa 2641 aaatatcccc cggcttgtga gtggatgggt aaaacctatc atcataggtc gtcatgctta 2701 tggggatcaa tacagagcaa ctgattttgt tgttcctggg cctggaaaag tagagataac 2761 ctacacacca agtgacggaa cccaaaaggt gacatacctg gtacataact ttgaagaagg 2821 tggtggtgtt gccatgggga tgtataatca agataagtca attgaagatt ttgcacacag 2881 ttccttccaa atggctctgt ctaagggttg gcctttgtat ctgagcacca aaaacactat 2941 tctgaagaaa tatgatgggc gttttaaaga catctttcag gagatatatg acaagcagta 3001 caagtcccag tttgaagctc aaaagatctg gtatgagcat aggctcatcg acgacatggt 3061 ggcccaagct atgaaatcag agggaggctt catctgggcc tgtaaaaact atgatggtga 3121 cgtgcagtcg gactctgtgg cccaagggta tggctctctc ggcatgatga ccagcgtgct 3181 ggtttgtcca gatggcaaga cagtagaagc agaggctgcc cacgggactg taacccgtca 3241 ctaccgcatg taccagaaag gacaggagac gtccaccaat cccattgctt ccatttttgc 3301 ctggaccaga gggttagccc acagagcaaa gcttgataac aataaagagc ttgccttctt 3361 tgcaaatgct ttggaagaag tctctattga gacaattgag gctggcttca tgaccaagga 3421 cttggctgct tgcattaaag gtttacccaa tgtgcaacgt tctgactact tgaatacatt 3481 tgagttcatg gataaacttg gagaaaactt gaagatcaaa ctagctcagg ccaaacttga 3541 ttacaaggat gacgacgata agtaaggatc cgcggccgca aggatctgcg atcgctccgg 3601 tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgg ggggaggggt 3661 cggcaattga acgggtgcct agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt 3721 gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg cagtagtcgc 3781 cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca cagctgaagc ttcgaggggc 3841 tcgcatctct ccttcacgcg cccgccgccc tacctgaggc cgccatccac gccggttgag 3901 tcgcgttctg ccgcctcccg cctgtggtgc ctcctgaact gcgtccgccg tctaggtaag 3961 tttaaagctc aggtcgagac cgggcctttg tccggcgctc ccttggagcc tacctagact 4021 cagccggctc tccacgcttt gcctgaccct gcttgctcaa ctctacgtct ttgtttcgtt 4081 ttctgttctg cgccgttaca gatccaagct gtgaccggcg cctacgctag acgccaccat 4141 ggagagcgac gagagcggcc tgcccgccat ggagatcgag tgccgcatca ccggcaccct 4201 gaacggcgtg gagttcgagc tggtgggcgg cggagagggc acccccaagc agggccgcat 4261 gaccaacaag atgaagagca ccaaaggcgc cctgaccttc agcccctacc tgctgagcca 4321 cgtgatgggc tacggcttct accacttcgg cacctacccc agcggctacg agaacccctt 4381 cctgcacgcc atcaacaacg gcggctacac caacacccgc atcgagaagt acgaggacgg 4441 cggcgtgctg cacgtgagct tcagctaccg ctacgaggcc ggccgcgtga tcggcgactt 4501 caaggtggtg ggcaccggct tccccgagga cagcgtgatc ttcaccgaca agatcatccg 4561 cagcaacgcc accgtggagc acctgcaccc catgggcgat aacgtgctgg tgggcagctt 4621 cgcccgcacc ttcagcctgc gcgacggcgg ctactacagc ttcgtggtgg acagccacat 4681 gcacttcaag agcgccatcc accccagcat cctgcagaac gggggcccca tgttcgcctt 4741 ccgccgcgtg gaggagctgc acagcaacac cgagctgggc atcgtggagt accagcacgc 4801 cttcaagacc cccatcgcct tcgccagatc ccgcgctcag tcgtccaatt ctgccgtgga 4861 cggcaccgcc ggacccggct ccaccggatc tcgcgagggc agaggaagtc ttctaacatg 4921 cggtgacgtg gaggagaatc ccggccctat gaccgagtac aagcccacgg tgcgcctcgc 4981 cacccgcgac gacgtcccca gggccgtacg caccctcgcc gccgcgttcg ccgactaccc 5041 cgccacgcgc cacaccgtcg atccggaccg ccacatcgag cgggtcaccg agctgcaaga 5101 actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc 5161 cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat 5221 cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaagg 5281 cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc 5341 gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggcggc 5401 cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc tccccttcta 5461 cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg 5521 gtgcatgacc cgcaagcccg gtgcctgaaa tcaacctctg gattacaaaa tttgtgaaag 5581 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat 5641 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc 5701 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg 5761 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct 5821 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct 5881 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg 5941 gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac 6001 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct 6061 gccggctctg cggcctcttc cgcgtctccg ccttcgccct cagacgagtc ggatctccct 6121 ttggccgcct ccccgcctgg tacctttaag accaatgact tacaaggcag ctgtagatct 6181 tagccacttt ttaaaagaaa aggggggact ggaagggcta attcactccc aacgaaaata 6241 agatctgctt tttgcttgta ctgggtctct ctggttagac cagatctgag cctgggagct 6301 ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca 6361 agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta 6421 gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat cttattattc agtatttata 6481 acttgcaaag aaatgaatat cagagagtga gaggaacttg tttattgcag cttataatgg 6541 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc 6601 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggctct agctatcccg 6661 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt 6721 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt 6781 tttggaggcc tagacttttg cagagacggc ccaaattcgt aatcatggtc atagctgttt 6841 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag 6901 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg 6961 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg 7021 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc 7081 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc 7141 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg 7201 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat 7261 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag 7321 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga 7381 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg 7441 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt 7501 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac 7561 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc 7621 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt 7681 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc 7741 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc 7801 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg 7861 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag 7921 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg 7981 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt 8041 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca 8101 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca 8161 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc 8221 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt 8281 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg 8341 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc 8401 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg 8461 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga 8521 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga 8581 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta 8641 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg 8701 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact 8761 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata 8821 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt 8881 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa 8941 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt 9001 atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc 9061 ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg 9121 taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt 9181 cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg 9241 tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgcca ttcgccattc 9301 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg 9361 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca 9421 cgacgttgta aaacgacggc cagtgccaag ctg //