LOCUS V045328 9584 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V045328 VERSION V045328 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..3585 /label="Runx3(NM_019732)" /note="Runx3(NM_019732)" /gene="Runx3" CDS 3592..3618 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" CDS 3622..3648 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" CDS 3652..3678 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" promoter 3743..3954 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3967..4235 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4264..4273 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4294..4956 /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 5026..5079 /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 5080..5679 /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 5680..6267 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6341..6574 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6646..6767 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6786..6921 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6959..6975) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6983..6999 /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(7007..7037) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7052..7073 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7361..7949) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8120..8980) /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(8981..9085) /gene="bla" /label="AmpR promoter" primer_bind 9559..9575 /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 gccaccatgg cttccaacag catctttgac 2341 tccttcccca actatacacc aaccttcata cgagacccga gcaccagccg ccgcttcact 2401 cccccctcca cggccttccc ctgcggcggc ggcggcggcg gcaagatggg cgagaacagc 2461 ggcgcgctaa gcgcgcaggc aaccgcgggc cccggcggcc gcacccggcc cgaagtgcgc 2521 tcgatggtgg acgtgctggc cgaccacgcg ggagagctcg tgcgcaccga cagccccaac 2581 ttcctctgct ccgtgctgcc ctcgcactgg cgctgcaaca agacgctgcc ggtcgccttc 2641 aaggtggtgg ccctggggga tgtgccggat ggaacggtgg tgaccgtgat ggccggcaat 2701 gatgagaact actccgccga gctgcgcaac gcttccgctg tcatgaagaa ccaagtggcc 2761 aggttcaacg accttcgatt cgtgggccgc agtgggcgag ggaagagttt cacgctcaca 2821 atcaccgtgt tcaccaaccc tacccaagtg gctacctacc accgagccat caaggtcact 2881 gtggatggac cccgggaacc ccgacggcac cggcagaaga tagaagacca gaccaaggcc 2941 ttccccgacc gctttggaga cctgcgcatg cgtgtaacac caagcacacc cagcccccgt 3001 ggctctctca gcaccacgag ccacttcagc agccaggccc agaccccaat ccaaggctcc 3061 tcagacctga accccttctc cgacccccgc cagtttgacc gctccttccc tacgctgcag 3121 agcctcacag agagccgctt cccggacccc aggatgcact acccgggagc catgtctgcc 3181 gccttcccct acagcgccac accatcgggc accagcctgg gcagcctgag cgtggcgggc 3241 atgccggcca gcagccgctt ccaccacacc tacctccctc cgccctaccc cggggcccca 3301 cagagccaga gcgggccctt tcaggccaac cccgcgccct accacctctt ttacggcgcc 3361 tcctccggct cctaccagtt ctccatggca gccgcgggag gtggtgagcg ctcgcccacc 3421 cgcatgctga cctcctgccc cagcggcgct tcggtgtcag caggcaacct catgaacccc 3481 agcctgggcc aggctgatgg cgtggaagcc gacggcagcc acagcaactc gcccacggcc 3541 ctgagcacgc cgggccgcat ggacgaggcc gtgtggcggc cctacctcga gtacccatac 3601 gacgtacctg attatgcaac ttatccttat gatgtacctg attatgctag ctatccttat 3661 gatgtcccag actacgcata gggatccctg cgatatccag cgcggccgcg aaggatctgc 3721 gatcgctccg gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc cgagaagttg 3781 gggggagggg tcggcaattg aacgggtgcc tagagaaggt ggcgcggggt aaactgggaa 3841 agtgatgtcg tgtactggct ccgccttttt cccgagggtg ggggagaacc gtatataagt 3901 gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg ccgccagaac acagctgaag 3961 cttcgagggg ctcgcatctc tccttcacgc gcccgccgcc ctacctgagg ccgccatcca 4021 cgccggttga gtcgcgttct gccgcctccc gcctgtggtg cctcctgaac tgcgtccgcc 4081 gtctaggtaa gtttaaagct caggtcgaga ccgggccttt gtccggcgct cccttggagc 4141 ctacctagac tcagccggct ctccacgctt tgcctgaccc tgcttgctca actctacgtc 4201 tttgtttcgt tttctgttct gcgccgttac agatccaagc tgtgaccggc gcctacgcta 4261 gacgccacca tggagagcga cgagagcggc ctgcccgcca tggagatcga gtgccgcatc 4321 accggcaccc tgaacggcgt ggagttcgag ctggtgggcg gcggagaggg cacccccaag 4381 cagggccgca tgaccaacaa gatgaagagc accaaaggcg ccctgacctt cagcccctac 4441 ctgctgagcc acgtgatggg ctacggcttc taccacttcg gcacctaccc cagcggctac 4501 gagaacccct tcctgcacgc catcaacaac ggcggctaca ccaacacccg catcgagaag 4561 tacgaggacg gcggcgtgct gcacgtgagc ttcagctacc gctacgaggc cggccgcgtg 4621 atcggcgact tcaaggtggt gggcaccggc ttccccgagg acagcgtgat cttcaccgac 4681 aagatcatcc gcagcaacgc caccgtggag cacctgcacc ccatgggcga taacgtgctg 4741 gtgggcagct tcgcccgcac cttcagcctg cgcgacggcg gctactacag cttcgtggtg 4801 gacagccaca tgcacttcaa gagcgccatc caccccagca tcctgcagaa cgggggcccc 4861 atgttcgcct tccgccgcgt ggaggagctg cacagcaaca ccgagctggg catcgtggag 4921 taccagcacg ccttcaagac ccccatcgcc ttcgccagat cccgcgctca gtcgtccaat 4981 tctgccgtgg acggcaccgc cggacccggc tccaccggat ctcgcgaggg cagaggaagt 5041 cttctaacat gcggtgacgt ggaggagaat cccggcccta tgaccgagta caagcccacg 5101 gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc 5161 gccgactacc ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc 5221 gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg 5281 gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg 5341 ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa 5401 cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc 5461 gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga 5521 gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac 5581 ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga 5641 ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgaa atcaacctct ggattacaaa 5701 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac 5761 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc 5821 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt 5881 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc 5941 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc 6001 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg 6061 gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt 6121 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc 6181 cgcggcctgc tgccggctct gcggcctctt ccgcgtctcc gccttcgccc tcagacgagt 6241 cggatctccc tttggccgcc tccccgcctg gtacctttaa gaccaatgac ttacaaggca 6301 gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc 6361 caacgaaaat aagatctgct ttttgcttgt actgggtctc tctggttaga ccagatctga 6421 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct 6481 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc 6541 agaccctttt agtcagtgtg gaaaatctct agcagtagta gttcatgtca tcttattatt 6601 cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggaactt gtttattgca 6661 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt 6721 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggctc 6781 tagctatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa 6841 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt 6901 gaggaggctt ttttggaggc ctagactttt gcagagacgg cccaaattcg taatcatggt 6961 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg 7021 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt 7081 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg 7141 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg 7201 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa 7261 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc 7321 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc 7381 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat 7441 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc 7501 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct 7561 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg 7621 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc 7681 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga 7741 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa 7801 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta 7861 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc 7921 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg 7981 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga 8041 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg 8101 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct 8161 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg 8221 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc 8281 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa 8341 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc 8401 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt 8461 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc 8521 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt 8581 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc 8641 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt 8701 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata 8761 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga 8821 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag 8881 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa 8941 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt 9001 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga 9061 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag 9121 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc 9181 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca 9241 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg 9301 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc 9361 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc 9421 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat 9481 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt 9541 tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa gctg //