LOCUS V041188 9256 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V041188 VERSION V041188 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..3282 /label="EHF(NM_001206616)" /note="EHF(NM_001206616)" /gene="EHF" CDS 3289..3354 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3398..3609 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3622..3890 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 3919..3928 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 3949..4611 /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 4681..4734 /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 4735..5334 /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 5335..5923 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5997..6230 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6302..6423 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6463..6598 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6631..6647) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6655..6671 /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(6679..6709) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6724..6745 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7033..7621) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7792..8652) /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(8653..8757) /gene="bla" /label="AmpR promoter" primer_bind 9231..9247 /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 ggttgccgga gagaagagga 2341 ttggtcctgc ttttaagcct agctgaaatt cttttcaaga tcatgattct ggaaggaggt 2401 ggtgtaatga atctcaaccc cggcaacaac ctccttcacc agccgccagc ctggacagac 2461 agctactcca cgtgcaatgt ttccagtggg ttttttggag gccagtggca tgaaattcat 2521 cctcagtact ggaccaagta ccaggtgtgg gagtggctcc agcacctcct ggacaccaac 2581 cagctggatg ccaattgtat ccctttccaa gagttcgaca tcaacggcga gcacctctgc 2641 agcatgagtt tgcaggagtt cacccgggcg gcagggacgg cggggcagct cctctacagc 2701 aacttgcagc atctgaagtg gaacggccag tgcagtagtg acctgttcca gtccacacac 2761 aatgtcattg tcaagactga acaaactgag ccttccatca tgaacacctg gaaagacgag 2821 aactatttat atgacaccaa ctatggtagc acagtagatt tgttggacag caaaactttc 2881 tgccgggctc agatctccat gacaaccacc agtcaccttc ctgttgcaga gtcacctgat 2941 atgaaaaagg agcaagaccc ccctgccaag tgccacacca aaaagcacaa cccgagaggg 3001 actcacttat gggaattcat ccgcgacatc ctcttgaacc cagacaagaa cccaggatta 3061 ataaaatggg aagaccgatc tgagggcgtc ttcaggttct tgaaatcaga ggcagtggct 3121 cagctatggg gtaaaaagaa gaacaacagc agcatgacct atgaaaagct cagccgagct 3181 atgagatatt actacaaaag agaaattctg gagcgtgtgg atggacgaag actggtatat 3241 aaatttggga agaatgcccg aggatggaga gaaaatgaaa acggatccga ctacaaagac 3301 catgacggtg attataaaga tcatgacatc gactacaagg atgacgatga caagtagcgg 3361 ccgcgaagga tctgcgatcg ctccggtgcc cgtcagtggg cagagcgcac atcgcccaca 3421 gtccccgaga agttgggggg aggggtcggc aattgaacgg gtgcctagag aaggtggcgc 3481 ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc tttttcccga gggtggggga 3541 gaaccgtata taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg gtttgccgcc 3601 agaacacagc tgaagcttcg aggggctcgc atctctcctt cacgcgcccg ccgccctacc 3661 tgaggccgcc atccacgccg gttgagtcgc gttctgccgc ctcccgcctg tggtgcctcc 3721 tgaactgcgt ccgccgtcta ggtaagttta aagctcaggt cgagaccggg cctttgtccg 3781 gcgctccctt ggagcctacc tagactcagc cggctctcca cgctttgcct gaccctgctt 3841 gctcaactct acgtctttgt ttcgttttct gttctgcgcc gttacagatc caagctgtga 3901 ccggcgccta cgctagacgc caccatggag agcgacgaga gcggcctgcc cgccatggag 3961 atcgagtgcc gcatcaccgg caccctgaac ggcgtggagt tcgagctggt gggcggcgga 4021 gagggcaccc ccaagcaggg ccgcatgacc aacaagatga agagcaccaa aggcgccctg 4081 accttcagcc cctacctgct gagccacgtg atgggctacg gcttctacca cttcggcacc 4141 taccccagcg gctacgagaa ccccttcctg cacgccatca acaacggcgg ctacaccaac 4201 acccgcatcg agaagtacga ggacggcggc gtgctgcacg tgagcttcag ctaccgctac 4261 gaggccggcc gcgtgatcgg cgacttcaag gtggtgggca ccggcttccc cgaggacagc 4321 gtgatcttca ccgacaagat catccgcagc aacgccaccg tggagcacct gcaccccatg 4381 ggcgataacg tgctggtggg cagcttcgcc cgcaccttca gcctgcgcga cggcggctac 4441 tacagcttcg tggtggacag ccacatgcac ttcaagagcg ccatccaccc cagcatcctg 4501 cagaacgggg gccccatgtt cgccttccgc cgcgtggagg agctgcacag caacaccgag 4561 ctgggcatcg tggagtacca gcacgccttc aagaccccca tcgccttcgc cagatcccgc 4621 gctcagtcgt ccaattctgc cgtggacggc accgccggac ccggctccac cggatctcgc 4681 gagggcagag gaagtcttct aacatgcggt gacgtggagg agaatcccgg ccctatgacc 4741 gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tccccagggc cgtacgcacc 4801 ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac 4861 atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc 4921 aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc 4981 gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg 5041 ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg 5101 tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc 5161 gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc 5221 tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc 5281 gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctgaaatcaa 5341 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt 5401 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct 5461 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc 5521 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg 5581 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc 5641 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc 5701 actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt 5761 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca 5821 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt 5881 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggtac ctttaagacc 5941 aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg ggggactgga 6001 agggctaatt cactcccaac gaagataaga tctgcttttt gcttgtactg ggtctctctg 6061 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc 6121 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg 6181 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gtagtagttc 6241 atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag agagtgagag 6301 gaacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac 6361 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc 6421 ttatcatgtc tggctctagc tatcccgccc ctaactccgc ccatcccgcc cctaactccg 6481 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc 6541 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta 6601 gacttttgca gagaccaaat tcgtaatcat gtcatagctg tttcctgtgt gaaattgtta 6661 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc 6721 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg 6781 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg 6841 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg 6901 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa 6961 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc 7021 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc 7081 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag 7141 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct 7201 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta 7261 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc 7321 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc 7381 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt 7441 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct 7501 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc 7561 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca 7621 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta 7681 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa 7741 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg 7801 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg 7861 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc 7921 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc 7981 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa 8041 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc 8101 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg 8161 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc 8221 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat 8281 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg 8341 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc 8401 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg 8461 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat 8521 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg 8581 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg 8641 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct 8701 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac 8761 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta 8821 taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa 8881 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag 8941 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta 9001 tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag 9061 atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg 9121 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc 9181 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac 9241 ggccagtgcc aagctg //