LOCUS V026538 9193 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V026538 VERSION V026538 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 2311..3219 /label="Cd47(NM_001368415)" /note="Cd47(NM_001368415)" /gene="Cd47" CDS 3226..3291 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3335..3546 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3559..3827 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 3856..3865 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 3886..4548 /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 4618..4671 /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 4672..5271 /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 5272..5860 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5934..6167 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6239..6360 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6400..6535 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6568..6584) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6592..6608 /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(6616..6646) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6661..6682 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6970..7558) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7729..8589) /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(8590..8694) /gene="bla" /label="AmpR promoter" primer_bind 9168..9184 /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 atgtggccct tggcggcggc gctgttgctg 2341 ggctcctgct gctgcggttc agctcaacta ctgtttagta acgtcaactc catagagttc 2401 acttcatgca atgaaactgt ggtcatccct tgcatcgtcc gtaatgtgga ggcgcaaagc 2461 accgaagaaa tgtttgtgaa gtggaagttg aacaaatcgt atattttcat ctatgatgga 2521 aataaaaata gcactactac agatcaaaac tttaccagtg caaaaatctc agtctcagac 2581 ttaatcaatg gcattgcctc tttgaaaatg gataagcgcg atgccatggt gggaaactac 2641 acttgcgaag tgacagagtt atccagagaa ggcaaaacag ttatagagct gaaaaaccgc 2701 acggtttcgt ggttttctcc aaatgaaaag atcctcattg ttattttccc aattttggct 2761 atactcctgt tctggggaaa gtttggtatt ttaacactca aatataaatc cagccatacg 2821 aataagagaa tcattctgct gctcgttgcc gggctggtgc tcacagtcat cgtggttgtt 2881 ggagccatcc ttctcatccc aggagaaaag cccgtgaaga atgcttctgg acttggcctc 2941 attgtaatct ctacggggat attaatacta cttcagtaca atgtgtttat gacagctttt 3001 ggaatgacct ctttcaccat tgccatattg atcactcaag tgctgggcta cgtccttgct 3061 ttggtcgggc tgtgtctctg catcatggca tgtgagccag tgcacggccc ccttttgatt 3121 tcaggtttgg ggatcatagc tctagcagaa ctacttggat tagtttatat gaagtttgtc 3181 gcttccaacc agaggactat ccaacctcct aggaataggg gatccgacta caaagaccat 3241 gacggtgatt ataaagatca tgacatcgac tacaaggatg acgatgacaa gtagcggccg 3301 cgaaggatct gcgatcgctc cggtgcccgt cagtgggcag agcgcacatc gcccacagtc 3361 cccgagaagt tggggggagg ggtcggcaat tgaacgggtg cctagagaag gtggcgcggg 3421 gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa 3481 ccgtatataa gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt tgccgccaga 3541 acacagctga agcttcgagg ggctcgcatc tctccttcac gcgcccgccg ccctacctga 3601 ggccgccatc cacgccggtt gagtcgcgtt ctgccgcctc ccgcctgtgg tgcctcctga 3661 actgcgtccg ccgtctaggt aagtttaaag ctcaggtcga gaccgggcct ttgtccggcg 3721 ctcccttgga gcctacctag actcagccgg ctctccacgc tttgcctgac cctgcttgct 3781 caactctacg tctttgtttc gttttctgtt ctgcgccgtt acagatccaa gctgtgaccg 3841 gcgcctacgc tagacgccac catggagagc gacgagagcg gcctgcccgc catggagatc 3901 gagtgccgca tcaccggcac cctgaacggc gtggagttcg agctggtggg cggcggagag 3961 ggcaccccca agcagggccg catgaccaac aagatgaaga gcaccaaagg cgccctgacc 4021 ttcagcccct acctgctgag ccacgtgatg ggctacggct tctaccactt cggcacctac 4081 cccagcggct acgagaaccc cttcctgcac gccatcaaca acggcggcta caccaacacc 4141 cgcatcgaga agtacgagga cggcggcgtg ctgcacgtga gcttcagcta ccgctacgag 4201 gccggccgcg tgatcggcga cttcaaggtg gtgggcaccg gcttccccga ggacagcgtg 4261 atcttcaccg acaagatcat ccgcagcaac gccaccgtgg agcacctgca ccccatgggc 4321 gataacgtgc tggtgggcag cttcgcccgc accttcagcc tgcgcgacgg cggctactac 4381 agcttcgtgg tggacagcca catgcacttc aagagcgcca tccaccccag catcctgcag 4441 aacgggggcc ccatgttcgc cttccgccgc gtggaggagc tgcacagcaa caccgagctg 4501 ggcatcgtgg agtaccagca cgccttcaag acccccatcg ccttcgccag atcccgcgct 4561 cagtcgtcca attctgccgt ggacggcacc gccggacccg gctccaccgg atctcgcgag 4621 ggcagaggaa gtcttctaac atgcggtgac gtggaggaga atcccggccc tatgaccgag 4681 tacaagccca cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt acgcaccctc 4741 gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgatccgga ccgccacatc 4801 gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag 4861 gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa 4921 gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg 4981 gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg 5041 ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc 5101 gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc 5161 gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag 5221 gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg aaatcaacct 5281 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg 5341 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc 5401 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt 5461 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc 5521 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg 5581 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact 5641 gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt 5701 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg 5761 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc 5821 cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggtacctt taagaccaat 5881 gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg 5941 gctaattcac tcccaacgaa gataagatct gctttttgct tgtactgggt ctctctggtt 6001 agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca 6061 ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa 6121 ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta gtagttcatg 6181 tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga gtgagaggaa 6241 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa 6301 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta 6361 tcatgtctgg ctctagctat cccgccccta actccgccca tcccgcccct aactccgccc 6421 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag 6481 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctagac 6541 ttttgcagag accaaattcg taatcatgtc atagctgttt cctgtgtgaa attgttatcc 6601 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta 6661 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa 6721 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat 6781 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg 6841 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc 6901 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt 6961 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag 7021 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc 7081 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc 7141 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt 7201 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt 7261 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc 7321 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa 7381 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa 7441 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg 7501 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga 7561 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg 7621 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg 7681 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt 7741 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact 7801 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat 7861 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg 7921 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg 7981 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat 8041 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc 8101 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt 8161 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc 8221 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga 8281 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc 8341 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa 8401 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta 8461 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg 8521 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg 8581 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat 8641 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt 8701 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa 8761 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct 8821 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag 8881 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc 8941 ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg 9001 cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggga 9061 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc 9121 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc 9181 cagtgccaag ctg //