LOCUS V041395 9182 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V041395 VERSION V041395 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..3019 /label="Tnfsf14(NM_019418)" /note="Tnfsf14(NM_019418)" /gene="Tnfsf14" CDS 3026..3091 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3135..3346 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3359..3627 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 3656..3665 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 3686..4348 /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 4418..4471 /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 4478..5272 /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 5279..5867 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5939..6172 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6244..6365 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6384..6519 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6557..6573) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6581..6597 /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(6605..6635) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6650..6671 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6959..7547) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7718..8578) /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(8579..8683) /gene="bla" /label="AmpR promoter" primer_bind 9157..9173 /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 ccatggagag tgtggtacag ccttcagtgt ttgtggtgga 2341 tggacagacg gacatcccat tcaggcggct ggaacagaac caccggagac ggcgctgtgg 2401 cactgtccag gtcagcctgg ccctggtgct gctgctaggt gctgggctgg ccactcaggg 2461 ctggtttctc ctgagactgc atcaacgtct tggagacata gtagctcatc tgccagatgg 2521 aggcaaaggc tcctgggaga agctgataca agatcaacga tctcaccagg ccaacccagc 2581 agcacatctt acaggagcca acgccagctt gataggtatt ggtggacctc tgttatggga 2641 gacacgactt ggcctggcct tcttgagggg cttgacgtat catgatgggg ccctggtgac 2701 catggagccc ggttactact atgtgtactc caaagtgcag ctgagcggcg tgggctgccc 2761 ccaggggctg gccaatggcc tccccatcac ccatggacta tacaagcgca catcccgcta 2821 cccgaaggag ttagaactgc tggtcagtcg gcggtcaccc tgtggccggg ccaacagctc 2881 ccgagtctgg tgggacagca gcttcctggg cggcgtggta catctggagg ctggggaaga 2941 ggtggtggtc cgcgtgcctg gaaaccgcct ggtcagacca cgtgacggca ccaggtccta 3001 tttcggagct ttcatggtcg gatccgacta caaagaccat gacggtgatt ataaagatca 3061 tgacatcgac tacaaggatg acgatgacaa gtagcggccg cgaaggatct gcgatcgctc 3121 cggtgcccgt cagtgggcag agcgcacatc gcccacagtc cccgagaagt tggggggagg 3181 ggtcggcaat tgaacgggtg cctagagaag gtggcgcggg gtaaactggg aaagtgatgt 3241 cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt 3301 cgccgtgaac gttctttttc gcaacgggtt tgccgccaga acacagctga agcttcgagg 3361 ggctcgcatc tctccttcac gcgcccgccg ccctacctga ggccgccatc cacgccggtt 3421 gagtcgcgtt ctgccgcctc ccgcctgtgg tgcctcctga actgcgtccg ccgtctaggt 3481 aagtttaaag ctcaggtcga gaccgggcct ttgtccggcg ctcccttgga gcctacctag 3541 actcagccgg ctctccacgc tttgcctgac cctgcttgct caactctacg tctttgtttc 3601 gttttctgtt ctgcgccgtt acagatccaa gctgtgaccg gcgcctacgc tagacgccac 3661 catggagagc gacgagagcg gcctgcccgc catggagatc gagtgccgca tcaccggcac 3721 cctgaacggc gtggagttcg agctggtggg cggcggagag ggcaccccca agcagggccg 3781 catgaccaac aagatgaaga gcaccaaagg cgccctgacc ttcagcccct acctgctgag 3841 ccacgtgatg ggctacggct tctaccactt cggcacctac cccagcggct acgagaaccc 3901 cttcctgcac gccatcaaca acggcggcta caccaacacc cgcatcgaga agtacgagga 3961 cggcggcgtg ctgcacgtga gcttcagcta ccgctacgag gccggccgcg tgatcggcga 4021 cttcaaggtg gtgggcaccg gcttccccga ggacagcgtg atcttcaccg acaagatcat 4081 ccgcagcaac gccaccgtgg agcacctgca ccccatgggc gataacgtgc tggtgggcag 4141 cttcgcccgc accttcagcc tgcgcgacgg cggctactac agcttcgtgg tggacagcca 4201 catgcacttc aagagcgcca tccaccccag catcctgcag aacgggggcc ccatgttcgc 4261 cttccgccgc gtggaggagc tgcacagcaa caccgagctg ggcatcgtgg agtaccagca 4321 cgccttcaag acccccatcg ccttcgccag atcccgcgct cagtcgtcca attctgccgt 4381 ggacggcacc gccggacccg gctccaccgg atctcgcgag ggcagaggaa gtcttctaac 4441 atgcggtgac gtggaggaga atcccggccc tgccaccatg gctgaacaag atggattgca 4501 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac 4561 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt 4621 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc 4681 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg 4741 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc 4801 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc 4861 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat 4921 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc 4981 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca 5041 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga 5101 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat 5161 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc 5221 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagtcgacaa 5281 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc 5341 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat 5401 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg 5461 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg 5521 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat 5581 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt 5641 gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc 5701 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa 5761 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg 5821 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcggt acctttaaga 5881 ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa ggggggactg 5941 gaagggctaa ttcactccca acgaagataa gatctgcttt ttgcttgtac tgggtctctc 6001 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag 6061 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct 6121 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtagtagt 6181 tcatgtcatc ttattattca gtatttataa cttgcaaaga aatgaatatc agagagtgag 6241 aggaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc 6301 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta 6361 tcttatcatg tctggctcta gctatcccgc ccctaactcc gcccagttcc gcccattctc 6421 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg 6481 agctattcca gaagtagtga ggaggctttt ttggaggcct agacttttgc agagacggcc 6541 caaattcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc 6601 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct 6661 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc 6721 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt 6781 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag 6841 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca 6901 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt 6961 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc 7021 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct 7081 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg 7141 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca 7201 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact 7261 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta 7321 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta 7381 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct 7441 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt 7501 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga 7561 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca 7621 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat 7681 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg 7741 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt 7801 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag 7861 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc 7921 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag 7981 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca 8041 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa 8101 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga 8161 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata 8221 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca 8281 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg 8341 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg 8401 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg 8461 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag 8521 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac 8581 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca 8641 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag 8701 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta 8761 tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc 8821 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc 8881 agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc 8941 agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa 9001 aataccgcat caggcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg 9061 tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa 9121 gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgccaagc 9181 tg //