LOCUS V026995 9908 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V026995 VERSION V026995 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..3909 /label="PKM(NM_002654)" /note="PKM(NM_002654)" /gene="PKM" CDS 3916..3942 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" CDS 3946..3972 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" CDS 3976..4002 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" promoter 4067..4278 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4291..4559 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4588..4597 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4618..5280 /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 5350..5403 /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 5404..6003 /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 6004..6591 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6665..6898 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6970..7091 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 7110..7245 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(7283..7299) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7307..7323 /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(7331..7361) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7376..7397 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7685..8273) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8444..9304) /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(9305..9409) /gene="bla" /label="AmpR promoter" primer_bind 9883..9899 /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 gccaccatgt cgaagcccca tagtgaagcc 2341 gggactgcct tcattcagac ccagcagctg cacgcagcca tggctgacac attcctggag 2401 cacatgtgcc gcctggacat tgattcacca cccatcacag cccggaacac tggcatcatc 2461 tgtaccattg gcccagcttc ccgatcagtg gagacgttga aggagatgat taagtctgga 2521 atgaatgtgg ctcgtctgaa cttctctcat ggaactcatg agtaccatgc ggagaccatc 2581 aagaatgtgc gcacagccac ggaaagcttt gcttctgacc ccatcctcta ccggcccgtt 2641 gctgtggctc tagacactaa aggacctgag atccgaactg ggctcatcaa gggcagcggc 2701 actgcagagg tggagctgaa gaagggagcc actctcaaaa tcacgctgga taacgcctac 2761 atggaaaagt gtgacgagaa catcctgtgg ctggactaca agaacatctg caaggtggtg 2821 gaagtgggca gcaagatcta cgtggatgat gggcttattt ctctccaggt gaagcagaaa 2881 ggtgccgact tcctggtgac ggaggtggaa aatggtggct ccttgggcag caagaagggt 2941 gtgaaccttc ctggggctgc tgtggacttg cctgctgtgt cggagaagga catccaggat 3001 ctgaagtttg gggtcgagca ggatgttgat atggtgtttg cgtcattcat ccgcaaggca 3061 tctgatgtcc atgaagttag gaaggtcctg ggagagaagg gaaagaacat caagattatc 3121 agcaaaatcg agaatcatga gggggttcgg aggtttgatg aaatcctgga ggccagtgat 3181 gggatcatgg tggctcgtgg tgatctaggc attgagattc ctgcagagaa ggtcttcctt 3241 gctcagaaga tgatgattgg acggtgcaac cgagctggga agcctgtcat ctgtgctact 3301 cagatgctgg agagcatgat caagaagccc cgccccactc gggctgaagg cagtgatgtg 3361 gccaatgcag tcctggatgg agccgactgc atcatgctgt ctggagaaac agccaaaggg 3421 gactatcctc tggaggctgt gcgcatgcag cacctgattg cccgtgaggc agaggctgcc 3481 atctaccact tgcaattatt tgaggaactc cgccgcctgg cgcccattac cagcgacccc 3541 acagaagcca ccgccgtggg tgccgtggag gcctccttca agtgctgcag tggggccata 3601 atcgtcctca ccaagtctgg caggtctgct caccaggtgg ccagataccg cccacgtgcc 3661 cccatcattg ctgtgacccg gaatccccag acagctcgtc aggcccacct gtaccgtggc 3721 atcttccctg tgctgtgcaa ggacccagtc caggaggcct gggctgagga cgtggacctc 3781 cgggtgaact ttgccatgaa tgttggcaag gcccgaggct tcttcaagaa gggagatgtg 3841 gtcattgtgc tgaccggatg gcgccctggc tccggcttca ccaacaccat gcgtgttgtt 3901 cctgtgccgc tcgagtaccc atacgacgta cctgattatg caacttatcc ttatgatgta 3961 cctgattatg ctagctatcc ttatgatgtc ccagactacg catagggatc cctgcgatat 4021 ccagcgcggc cgcgaaggat ctgcgatcgc tccggtgccc gtcagtgggc agagcgcaca 4081 tcgcccacag tccccgagaa gttgggggga ggggtcggca attgaacggg tgcctagaga 4141 aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag 4201 ggtgggggag aaccgtatat aagtgcagta gtcgccgtga acgttctttt tcgcaacggg 4261 tttgccgcca gaacacagct gaagcttcga ggggctcgca tctctccttc acgcgcccgc 4321 cgccctacct gaggccgcca tccacgccgg ttgagtcgcg ttctgccgcc tcccgcctgt 4381 ggtgcctcct gaactgcgtc cgccgtctag gtaagtttaa agctcaggtc gagaccgggc 4441 ctttgtccgg cgctcccttg gagcctacct agactcagcc ggctctccac gctttgcctg 4501 accctgcttg ctcaactcta cgtctttgtt tcgttttctg ttctgcgccg ttacagatcc 4561 aagctgtgac cggcgcctac gctagacgcc accatggaga gcgacgagag cggcctgccc 4621 gccatggaga tcgagtgccg catcaccggc accctgaacg gcgtggagtt cgagctggtg 4681 ggcggcggag agggcacccc caagcagggc cgcatgacca acaagatgaa gagcaccaaa 4741 ggcgccctga ccttcagccc ctacctgctg agccacgtga tgggctacgg cttctaccac 4801 ttcggcacct accccagcgg ctacgagaac cccttcctgc acgccatcaa caacggcggc 4861 tacaccaaca cccgcatcga gaagtacgag gacggcggcg tgctgcacgt gagcttcagc 4921 taccgctacg aggccggccg cgtgatcggc gacttcaagg tggtgggcac cggcttcccc 4981 gaggacagcg tgatcttcac cgacaagatc atccgcagca acgccaccgt ggagcacctg 5041 caccccatgg gcgataacgt gctggtgggc agcttcgccc gcaccttcag cctgcgcgac 5101 ggcggctact acagcttcgt ggtggacagc cacatgcact tcaagagcgc catccacccc 5161 agcatcctgc agaacggggg ccccatgttc gccttccgcc gcgtggagga gctgcacagc 5221 aacaccgagc tgggcatcgt ggagtaccag cacgccttca agacccccat cgccttcgcc 5281 agatcccgcg ctcagtcgtc caattctgcc gtggacggca ccgccggacc cggctccacc 5341 ggatctcgcg agggcagagg aagtcttcta acatgcggtg acgtggagga gaatcccggc 5401 cctatgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc 5461 gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgatccg 5521 gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc 5581 gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg 5641 gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc 5701 ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag 5761 gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg 5821 ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc 5881 ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc 5941 gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc 6001 tgaaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt 6061 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc 6121 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag 6181 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc 6241 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc 6301 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg 6361 ctgttgggca ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt tccatggctg 6421 ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc 6481 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt 6541 ctccgccttc gccctcagac gagtcggatc tccctttggc cgcctccccg cctggtacct 6601 ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg 6661 ggactggaag ggctaattca ctcccaacga aaataagatc tgctttttgc ttgtactggg 6721 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg 6781 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt 6841 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt 6901 agtagttcat gtcatcttat tattcagtat ttataacttg caaagaaatg aatatcagag 6961 agtgagagga acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca 7021 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc 7081 aatgtatctt atcatgtctg gctctagcta tcccgcccct aactccgccc agttccgccc 7141 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg 7201 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctagac ttttgcagag 7261 acggcccaaa ttcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca 7321 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag 7381 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt 7441 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc 7501 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg 7561 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa 7621 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg 7681 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga 7741 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg 7801 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg 7861 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc 7921 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg 7981 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca 8041 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt 8101 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag 8161 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg 8221 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc 8281 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt 8341 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt 8401 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca 8461 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg 8521 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac 8581 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg 8641 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc 8701 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta 8761 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac 8821 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc 8881 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac 8941 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact 9001 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa 9061 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt 9121 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca 9181 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa 9241 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac 9301 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg 9361 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc 9421 gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata 9481 ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac 9541 acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag 9601 cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat 9661 cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa 9721 ggagaaaata ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc 9781 gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc 9841 gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg 9901 ccaagctg //