LOCUS V043304 9673 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V043304 VERSION V043304 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 2288..2314 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" CDS 2318..2344 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" CDS 2348..2374 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" CDS 2381..3781 /label="PPP2R5E(NM_001282179)" /note="PPP2R5E(NM_001282179)" /gene="PPP2R5E" promoter 3832..4043 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4056..4324 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4353..4362 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4383..5045 /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 5115..5168 /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 5169..5768 /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 5769..6356 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6430..6663 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6735..6856 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6875..7010 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(7048..7064) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7072..7088 /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(7096..7126) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7141..7162 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7450..8038) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8209..9069) /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(9070..9174) /gene="bla" /label="AmpR promoter" primer_bind 9648..9664 /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 caccatgtac ccatacgacg tacctgatta tgcaacttat ccttatgatg tacctgatta 2341 tgctagctat ccttatgatg tcccagacta cgcagaattc atgtcctcag caccaactac 2401 tcctccatca gtggataaag tagacggatt ttctcggaag tccgtcagaa aagccagaca 2461 gaagaggtcg caaagttcct cacagtttag gtctcaaggc aagcctattg agttaacacc 2521 tctgccgctg ctaaaagacg ttccatcctc agagcagcct gaactgttcc taaagaaact 2581 tcagcagtgc tgtgtcattt ttgacttcat ggacacgcta tctgatctta aaatgaaaga 2641 atacaagcgc tccactctta atgaactggt ggactacatt acaataagca gaggctgttt 2701 gacagagcag acttaccctg aagtagttag aatggtatct tgcaatatat tcagaactct 2761 ccctcctagt gacagcaatg aatttgatcc agaagaagat gaacctaccc ttgaggcatc 2821 gtggccacac ttacagcttg tatatgaatt tttcatacga tttttggaaa gccaagaatt 2881 ccaacccagc attgccaaaa aatatataga tcagaaattt gtattacagc ttctggagct 2941 atttgacagc gaagaccctc gggaacggga ctacttaaaa acagtcttac acagaattta 3001 tggcaagttt cttggtctta gagcatttat ccgaaaacag attaacaata tttttctaag 3061 gtttgtttat gaaacagaac acttcaatgg tgtagctgaa ctgctggaaa tattaggaag 3121 tattatcaat ggctttgctt tacctcttaa ggcagaacac aaacagtttc tggtgaaagt 3181 attgatccct ttacacactg tcaggagctt atcactcttc catgcacagc tggcatattg 3241 tatagtacag tttctggaga aagatccttc actcacagaa ccagttatta gggggttaat 3301 gaaattttgg cctaaaacat gtagtcaaaa agaggtcatg ttccttgggg aactggaaga 3361 aatattggat gtgattgaac cttcacaatt tgttaaaatc caagaacctt tgtttaaaca 3421 aatcgccaag tgtgtatcta gcccccattt tcaggtggca gaaagagcac tctattattg 3481 gaataatgaa tacatcatga gtttgataga agaaaactct aacgtcatcc ttcccatcat 3541 gttttccagc ctttatagga tttcaaaaga acattggaat ccggctattg tggcgttggt 3601 gtacaatgtg ttgaaggcat ttatggaaat gaacagcacc atgtttgacg agctgacagc 3661 cacatacaag tcagatcgtc agcgtgagaa aaagaaagaa aaggagcgtg aagaattgtg 3721 gaaaaaattg gaggatctgg agttaaagag aggtcttaga cgtgatggaa taattccaac 3781 ttaaggatcc gcggccgcga aggatctgcg atcgctccgg tgcccgtcag tgggcagagc 3841 gcacatcgcc cacagtcccc gagaagttgg ggggaggggt cggcaattga acgggtgcct 3901 agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt gtactggctc cgcctttttc 3961 ccgagggtgg gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca 4021 acgggtttgc cgccagaaca cagctgaagc ttcgaggggc tcgcatctct ccttcacgcg 4081 cccgccgccc tacctgaggc cgccatccac gccggttgag tcgcgttctg ccgcctcccg 4141 cctgtggtgc ctcctgaact gcgtccgccg tctaggtaag tttaaagctc aggtcgagac 4201 cgggcctttg tccggcgctc ccttggagcc tacctagact cagccggctc tccacgcttt 4261 gcctgaccct gcttgctcaa ctctacgtct ttgtttcgtt ttctgttctg cgccgttaca 4321 gatccaagct gtgaccggcg cctacgctag acgccaccat ggagagcgac gagagcggcc 4381 tgcccgccat ggagatcgag tgccgcatca ccggcaccct gaacggcgtg gagttcgagc 4441 tggtgggcgg cggagagggc acccccaagc agggccgcat gaccaacaag atgaagagca 4501 ccaaaggcgc cctgaccttc agcccctacc tgctgagcca cgtgatgggc tacggcttct 4561 accacttcgg cacctacccc agcggctacg agaacccctt cctgcacgcc atcaacaacg 4621 gcggctacac caacacccgc atcgagaagt acgaggacgg cggcgtgctg cacgtgagct 4681 tcagctaccg ctacgaggcc ggccgcgtga tcggcgactt caaggtggtg ggcaccggct 4741 tccccgagga cagcgtgatc ttcaccgaca agatcatccg cagcaacgcc accgtggagc 4801 acctgcaccc catgggcgat aacgtgctgg tgggcagctt cgcccgcacc ttcagcctgc 4861 gcgacggcgg ctactacagc ttcgtggtgg acagccacat gcacttcaag agcgccatcc 4921 accccagcat cctgcagaac gggggcccca tgttcgcctt ccgccgcgtg gaggagctgc 4981 acagcaacac cgagctgggc atcgtggagt accagcacgc cttcaagacc cccatcgcct 5041 tcgccagatc ccgcgctcag tcgtccaatt ctgccgtgga cggcaccgcc ggacccggct 5101 ccaccggatc tcgcgagggc agaggaagtc ttctaacatg cggtgacgtg gaggagaatc 5161 ccggccctat gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca 5221 gggccgtacg caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg 5281 atccggaccg ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg 5341 ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca 5401 cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc atggccgagt 5461 tgagcggttc ccggctggcc gcgcagcaac agatggaagg cctcctggcg ccgcaccggc 5521 ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac cagggcaagg 5581 gtctgggcag cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg 5641 ccttcctgga gacctccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg 5701 tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg 5761 gtgcctgaaa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact 5821 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg 5881 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg 5941 aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa 6001 cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc 6061 ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg 6121 ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaagctgacg tcctttccat 6181 ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt 6241 cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc 6301 cgcgtctccg ccttcgccct cagacgagtc ggatctccct ttggccgcct ccccgcctgg 6361 tacctttaag accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa 6421 aggggggact ggaagggcta attcactccc aacgaaaata agatctgctt tttgcttgta 6481 ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc 6541 cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt 6601 tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta 6661 gcagtagtag ttcatgtcat cttattattc agtatttata acttgcaaag aaatgaatat 6721 cagagagtga gaggaacttg tttattgcag cttataatgg ttacaaataa agcaatagca 6781 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac 6841 tcatcaatgt atcttatcat gtctggctct agctatcccg cccctaactc cgcccagttc 6901 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc 6961 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc tagacttttg 7021 cagagacggc ccaaattcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc 7081 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta 7141 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa 7201 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat 7261 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg 7321 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc 7381 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt 7441 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag 7501 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc 7561 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc 7621 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt 7681 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt 7741 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc 7801 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa 7861 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa 7921 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg 7981 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga 8041 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg 8101 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg 8161 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt 8221 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact 8281 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat 8341 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg 8401 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg 8461 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat 8521 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc 8581 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt 8641 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc 8701 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga 8761 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc 8821 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa 8881 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta 8941 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg 9001 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg 9061 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat 9121 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt 9181 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa 9241 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct 9301 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag 9361 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc 9421 ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg 9481 cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggga 9541 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc 9601 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc 9661 cagtgccaag ctg //