LOCUS V029279 9316 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V029279 VERSION V029279 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..3342 /label="Fam169b(NM_001374636)" /note="Fam169b(NM_001374636)" /gene="Fam169b" CDS 3349..3414 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3458..3669 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3682..3950 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 3979..3988 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4009..4671 /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 4741..4794 /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 4795..5394 /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 5395..5983 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6057..6290 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6362..6483 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6523..6658 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6691..6707) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6715..6731 /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(6739..6769) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6784..6805 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7093..7681) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7852..8712) /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(8713..8817) /gene="bla" /label="AmpR promoter" primer_bind 9291..9307 /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 aggctgaggc agtccgcgga 2341 gtcacagagg cttaccctgt ggacatcctg gaggatgacg cggagggcta tcaggcagca 2401 gccgaggctt actatgagat gctcagggaa ggagcccaga cctcagcaga ggttatctcc 2461 ctttccactg gggagcaggt cagacttgag acttcatctc tttgtttttg cacgatatac 2521 cgtgatgaac ctcaacataa aatactgggt ttggttaatc cccaggacac aaagacagtt 2581 gtggctgttt acttaaagga atcctggtgg tccatcgaag acatcctaag aacttctgat 2641 cccacgcggg aagggctcat gaaggttcag tcctttgggg aaaggattgt tctctttgtt 2701 ctcaacgtca ttgtttttgg aagactggag agaaggctgc acatcgatga catgtttttc 2761 ttacctcacc ctgcgaagga gcaagctaag attctgtgga aagatggagc cgcggttgca 2821 ttttactcag tcaaaatgaa aggcagcctc tgtggtgatg gtaccggcac atgctacctg 2881 ctgcctgtct tggacaccgt gtttgtgagg aggaagaacc gttgccaagg cctggggaca 2941 gccatgctga gggacttctg tgacaccttt caaggagacg aagctctggg tatcagttgt 3001 ccgatatctc ctgcaatgta cagagtcctc aggcagttcc tgttgacctg tccaggcgag 3061 agagggcgct tgtgggaggt ggagcccccg ggggcctggg gccagcagcg tgttaacatc 3121 tggctcaagg tttatcttca ggagcggaga cttcaagatg ggagcaccgt gcaccctaaa 3181 tgttctgaag aggacacgga cactcccggg caggcttctc aggaggatgg acccacacag 3241 tttaatcatg gagaaagcca caaggagtgg gctgttggag aaccggagag gacacagaat 3301 ggccgacgct gtgcgcaagt gtgtgaggaa gcgcgtcagg tgggatccga ctacaaagac 3361 catgacggtg attataaaga tcatgacatc gactacaagg atgacgatga caagtagcgg 3421 ccgcgaagga tctgcgatcg ctccggtgcc cgtcagtggg cagagcgcac atcgcccaca 3481 gtccccgaga agttgggggg aggggtcggc aattgaacgg gtgcctagag aaggtggcgc 3541 ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc tttttcccga gggtggggga 3601 gaaccgtata taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg gtttgccgcc 3661 agaacacagc tgaagcttcg aggggctcgc atctctcctt cacgcgcccg ccgccctacc 3721 tgaggccgcc atccacgccg gttgagtcgc gttctgccgc ctcccgcctg tggtgcctcc 3781 tgaactgcgt ccgccgtcta ggtaagttta aagctcaggt cgagaccggg cctttgtccg 3841 gcgctccctt ggagcctacc tagactcagc cggctctcca cgctttgcct gaccctgctt 3901 gctcaactct acgtctttgt ttcgttttct gttctgcgcc gttacagatc caagctgtga 3961 ccggcgccta cgctagacgc caccatggag agcgacgaga gcggcctgcc cgccatggag 4021 atcgagtgcc gcatcaccgg caccctgaac ggcgtggagt tcgagctggt gggcggcgga 4081 gagggcaccc ccaagcaggg ccgcatgacc aacaagatga agagcaccaa aggcgccctg 4141 accttcagcc cctacctgct gagccacgtg atgggctacg gcttctacca cttcggcacc 4201 taccccagcg gctacgagaa ccccttcctg cacgccatca acaacggcgg ctacaccaac 4261 acccgcatcg agaagtacga ggacggcggc gtgctgcacg tgagcttcag ctaccgctac 4321 gaggccggcc gcgtgatcgg cgacttcaag gtggtgggca ccggcttccc cgaggacagc 4381 gtgatcttca ccgacaagat catccgcagc aacgccaccg tggagcacct gcaccccatg 4441 ggcgataacg tgctggtggg cagcttcgcc cgcaccttca gcctgcgcga cggcggctac 4501 tacagcttcg tggtggacag ccacatgcac ttcaagagcg ccatccaccc cagcatcctg 4561 cagaacgggg gccccatgtt cgccttccgc cgcgtggagg agctgcacag caacaccgag 4621 ctgggcatcg tggagtacca gcacgccttc aagaccccca tcgccttcgc cagatcccgc 4681 gctcagtcgt ccaattctgc cgtggacggc accgccggac ccggctccac cggatctcgc 4741 gagggcagag gaagtcttct aacatgcggt gacgtggagg agaatcccgg ccctatgacc 4801 gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tccccagggc cgtacgcacc 4861 ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac 4921 atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc 4981 aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc 5041 gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg 5101 ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg 5161 tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc 5221 gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc 5281 tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc 5341 gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctgaaatcaa 5401 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt 5461 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct 5521 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc 5581 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg 5641 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc 5701 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc 5761 actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt 5821 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca 5881 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt 5941 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggtac ctttaagacc 6001 aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg ggggactgga 6061 agggctaatt cactcccaac gaagataaga tctgcttttt gcttgtactg ggtctctctg 6121 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc 6181 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg 6241 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gtagtagttc 6301 atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag agagtgagag 6361 gaacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac 6421 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc 6481 ttatcatgtc tggctctagc tatcccgccc ctaactccgc ccatcccgcc cctaactccg 6541 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc 6601 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta 6661 gacttttgca gagaccaaat tcgtaatcat gtcatagctg tttcctgtgt gaaattgtta 6721 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc 6781 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg 6841 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg 6901 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg 6961 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa 7021 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc 7081 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc 7141 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag 7201 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct 7261 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta 7321 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc 7381 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc 7441 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt 7501 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct 7561 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc 7621 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca 7681 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta 7741 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa 7801 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg 7861 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg 7921 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc 7981 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc 8041 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa 8101 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc 8161 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg 8221 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc 8281 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat 8341 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg 8401 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc 8461 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg 8521 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat 8581 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg 8641 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg 8701 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct 8761 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac 8821 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta 8881 taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa 8941 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag 9001 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta 9061 tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag 9121 atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg 9181 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc 9241 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac 9301 ggccagtgcc aagctg //