LOCUS V033258 9704 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V033258 VERSION V033258 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 2303..3541 /label="E2F4(NM_001950)" /note="E2F4(NM_001950)" /gene="E2F4" CDS 3548..3613 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3657..3868 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3881..4149 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4178..4187 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4208..4870 /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 4940..4993 /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 5000..5794 /codon_start=1 /gene="aph(3')-II (or nptII)" /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin®)" /product="aminoglycoside phosphotransferase from Tn5" /transl_table=1 /translation="MAEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*" /label="NeoR/KanR" misc_feature 5801..6389 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6461..6694 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6766..6887 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6906..7041 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(7079..7095) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7103..7119 /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(7127..7157) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7172..7193 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7481..8069) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8240..9100) /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(9101..9205) /gene="bla" /label="AmpR promoter" primer_bind 9679..9695 /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 gaattcgcca ccatggcgga ggccgggcca caggcgccgc cgcccccggg 2341 cactccaagc cggcacgaaa agagcctggg actgctcacc accaagttcg tgtcccttct 2401 gcaggaggcc aaggacggcg tgcttgacct caagctggca gctgacaccc tagctgtacg 2461 ccagaagcgg cggatttacg acattaccaa tgttttggaa ggtatcgggc taatcgagaa 2521 aaagtccaag aacagcatcc agtggaaggg tgtggggcct ggctgcaata cccgggagat 2581 tgctgacaaa ctgattgagc tcaaggcaga gatcgaggag ctgcagcagc gggagcaaga 2641 actagaccag cacaaggtgt gggtgcagca gagcatccgg aacgtcacag aggacgtgca 2701 gaacagctgt ttggcctacg tcactcatga ggacatctgc agatgctttg ctggagatac 2761 cctcttggcc atccgggccc catcaggcac cagcctggag gtgcccatcc cagagggtct 2821 caatgggcag aagaagtacc agattcacct gaagagtgtg agtggtccca ttgaggttct 2881 gctggtgaac aaggaggcat ggagctcacc ccctgtggct gtgcctgtgc caccacctga 2941 agatttgctc cagagcccat ctgctgtttc tacacctcca cctctgccca agcctgccct 3001 agcccagtcc caggaagcct cacgtccaaa tagtcctcag ctcactccca ctgctgtccc 3061 tggcagtgca gaagtccagg gaatggctgg cccagcagct gagatcacag tgagtggcgg 3121 ccctgggact gatagcaagg acagtggtga gctcagttca ctcccactgg gcccaacaac 3181 actggacacc cggccactgc agtcttctgc cctgctggac agcagcagca gcagcagcag 3241 cagcagcagc agcagcagca acagtaacag cagcagttcg tccggaccca acccttctac 3301 ctcctttgag cccatcaagg cagaccccac aggtgttttg gaactcccca aagagctgtc 3361 agaaatcttt gatcccacac gagagtgcat gagctcggag ctgctggagg agttgatgtc 3421 ctcagaagtg tttgcccctc tgcttcgtct ttctccaccc ccgggagacc acgattatat 3481 ctacaacctg gacgagagtg aaggtgtctg tgacctcttt gatgtgcctg ttctcaacct 3541 cggatccgac tacaaagacc atgacggtga ttataaagat catgacatcg actacaagga 3601 tgacgatgac aagtagcggc cgcgaaggat ctgcgatcgc tccggtgccc gtcagtgggc 3661 agagcgcaca tcgcccacag tccccgagaa gttgggggga ggggtcggca attgaacggg 3721 tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct 3781 ttttcccgag ggtgggggag aaccgtatat aagtgcagta gtcgccgtga acgttctttt 3841 tcgcaacggg tttgccgcca gaacacagct gaagcttcga ggggctcgca tctctccttc 3901 acgcgcccgc cgccctacct gaggccgcca tccacgccgg ttgagtcgcg ttctgccgcc 3961 tcccgcctgt ggtgcctcct gaactgcgtc cgccgtctag gtaagtttaa agctcaggtc 4021 gagaccgggc ctttgtccgg cgctcccttg gagcctacct agactcagcc ggctctccac 4081 gctttgcctg accctgcttg ctcaactcta cgtctttgtt tcgttttctg ttctgcgccg 4141 ttacagatcc aagctgtgac cggcgcctac gctagacgcc accatggaga gcgacgagag 4201 cggcctgccc gccatggaga tcgagtgccg catcaccggc accctgaacg gcgtggagtt 4261 cgagctggtg ggcggcggag agggcacccc caagcagggc cgcatgacca acaagatgaa 4321 gagcaccaaa ggcgccctga ccttcagccc ctacctgctg agccacgtga tgggctacgg 4381 cttctaccac ttcggcacct accccagcgg ctacgagaac cccttcctgc acgccatcaa 4441 caacggcggc tacaccaaca cccgcatcga gaagtacgag gacggcggcg tgctgcacgt 4501 gagcttcagc taccgctacg aggccggccg cgtgatcggc gacttcaagg tggtgggcac 4561 cggcttcccc gaggacagcg tgatcttcac cgacaagatc atccgcagca acgccaccgt 4621 ggagcacctg caccccatgg gcgataacgt gctggtgggc agcttcgccc gcaccttcag 4681 cctgcgcgac ggcggctact acagcttcgt ggtggacagc cacatgcact tcaagagcgc 4741 catccacccc agcatcctgc agaacggggg ccccatgttc gccttccgcc gcgtggagga 4801 gctgcacagc aacaccgagc tgggcatcgt ggagtaccag cacgccttca agacccccat 4861 cgccttcgcc agatcccgcg ctcagtcgtc caattctgcc gtggacggca ccgccggacc 4921 cggctccacc ggatctcgcg agggcagagg aagtcttcta acatgcggtg acgtggagga 4981 gaatcccggc cctgccacca tggctgaaca agatggattg cacgcaggtt ctccggccgc 5041 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc 5101 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc 5161 cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg 5221 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt 5281 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc 5341 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga 5401 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga 5461 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct 5521 caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc 5581 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt 5641 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg 5701 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat 5761 cgccttctat cgccttcttg acgagttctt ctgagtcgac aatcaacctc tggattacaa 5821 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata 5881 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc 5941 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg 6001 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac 6061 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat 6121 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt 6181 ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat 6241 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc 6301 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag 6361 tcggatctcc ctttgggccg cctccccgcg gtacctttaa gaccaatgac ttacaaggca 6421 gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc 6481 caacgaagat aagatctgct ttttgcttgt actgggtctc tctggttaga ccagatctga 6541 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct 6601 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc 6661 agaccctttt agtcagtgtg gaaaatctct agcagtagta gttcatgtca tcttattatt 6721 cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggaactt gtttattgca 6781 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt 6841 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggctc 6901 tagctatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa 6961 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt 7021 gaggaggctt ttttggaggc ctagactttt gcagagacgg cccaaattcg taatcatggt 7081 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg 7141 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt 7201 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg 7261 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg 7321 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa 7381 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc 7441 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc 7501 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat 7561 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc 7621 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct 7681 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg 7741 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc 7801 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga 7861 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa 7921 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta 7981 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc 8041 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg 8101 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga 8161 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg 8221 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct 8281 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg 8341 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc 8401 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa 8461 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc 8521 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt 8581 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc 8641 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt 8701 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc 8761 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt 8821 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata 8881 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga 8941 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag 9001 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa 9061 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt 9121 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga 9181 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag 9241 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc 9301 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca 9361 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg 9421 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc 9481 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc 9541 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat 9601 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt 9661 tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa gctg //