LOCUS V036113 9787 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V036113 VERSION V036113 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 2311..3813 /label="ARMC6(NM_001199196)" /note="ARMC6(NM_001199196)" /gene="ARMC6" CDS 3820..3885 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3929..4140 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4153..4421 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4450..4459 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4480..5142 /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 5212..5265 /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 5266..5865 /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 5866..6454 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6528..6761 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6833..6954 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6994..7129 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(7162..7178) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7186..7202 /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(7210..7240) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7255..7276 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7564..8152) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8323..9183) /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(9184..9288) /gene="bla" /label="AmpR promoter" primer_bind 9762..9778 /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 atgagtgaac gatgttgctc tagatacagc 2341 tcaggagcat ctatcggctg cacgccaaca tcaacacagg cgaagatggt ctccaagcgc 2401 attgcccagg agacctttga tgcagctgtg cgcgagaaca tcgaggagtt tgcgatgggg 2461 ccagaggagg cagtgaaaga ggccgtggag cagtttgaat cgcaaggggt tgatctgagc 2521 aacattgtaa agacggcacc taaagtctct gcagacggat cccaggagcc cacacatgac 2581 atcctgcaga tgctcagtga cctccaggag tctgtggcca gctctcgccc ccaggaggtg 2641 tcagcatacc tcacccgctt ctgcgaccag tgcaaacagg acaaggcctg ccgcttcctc 2701 gcggcccaga agggggccta ccccatcatc ttcactgcct ggaagctggc cactgcaggt 2761 gaccagggcc ttctgctcca gtccctcaat gccctgtcgg tgctgactga tggacagcca 2821 gacctcctgg atgcccaggg cctgcagctc ctagtggcca cgctgaccca gaatgctgat 2881 gaggctgacc tgacctgctc tgggatccgc tgtgtgcgtc acgcttgcct gaaacatgaa 2941 cagaatcggc aagacctggt gaaagctggc gtgctgcctc tgctgactgg tgccatcacc 3001 catcatggcc accacactga cgtggtcagg gaagcctgct gggccctgcg tgtcatgacc 3061 ttcgatgacg acatccgtgt gccctttggc catgcccaca accatgccaa gatgattgtg 3121 caggagaaca aaggcttgaa ggtgctcatc gaagccacca aagcgttcct ggataaccct 3181 ggcatcctga gcgagctctg tggaaccctg tcccgcctgg ccattcgcaa cgagttctgc 3241 caggaggtcg tcgacctcgg gggcctgagc attctggtgt ccctgctagc cgactgcaat 3301 gaccaccaga tgagggacca gagcggcgtt caggagctcg tgaagcaagt gctgagcacc 3361 ctgcgagcca tcgcaggcaa cgacgacgtg aaagatgcta ttgtccgtgc tggtgggacg 3421 gagtccatcg tggctgctat gacccagcat ctgaccagcc cccaggtgtg tgagcagagc 3481 tgcgcggccc tgtgcttcct ggccctgcgt aagcccgaca acagccgcat catcgtggag 3541 ggtggcgggg ctgtggcagc actgcaggcc atgaaggcac acccgcagaa ggccggcgtg 3601 cagaaacagg cttgcatgct gatccgaaac ctggtggccc acggccaggc cttctcgaag 3661 cccatcctgg acctgggggc tgaggcactc atcatgcagg cccgatctgc ccaccgtgac 3721 tgtgaggacg tggccaaggc cgccctgcgg gacctgggtt gtcatgtcga gctccgagag 3781 ctgtggacag gccagagggg caacctggcg ccaggatccg actacaaaga ccatgacggt 3841 gattataaag atcatgacat cgactacaag gatgacgatg acaagtagcg gccgcgaagg 3901 atctgcgatc gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag 3961 aagttggggg gaggggtcgg caattgaacg ggtgcctaga gaaggtggcg cggggtaaac 4021 tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat 4081 ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag 4141 ctgaagcttc gaggggctcg catctctcct tcacgcgccc gccgccctac ctgaggccgc 4201 catccacgcc ggttgagtcg cgttctgccg cctcccgcct gtggtgcctc ctgaactgcg 4261 tccgccgtct aggtaagttt aaagctcagg tcgagaccgg gcctttgtcc ggcgctccct 4321 tggagcctac ctagactcag ccggctctcc acgctttgcc tgaccctgct tgctcaactc 4381 tacgtctttg tttcgttttc tgttctgcgc cgttacagat ccaagctgtg accggcgcct 4441 acgctagacg ccaccatgga gagcgacgag agcggcctgc ccgccatgga gatcgagtgc 4501 cgcatcaccg gcaccctgaa cggcgtggag ttcgagctgg tgggcggcgg agagggcacc 4561 cccaagcagg gccgcatgac caacaagatg aagagcacca aaggcgccct gaccttcagc 4621 ccctacctgc tgagccacgt gatgggctac ggcttctacc acttcggcac ctaccccagc 4681 ggctacgaga accccttcct gcacgccatc aacaacggcg gctacaccaa cacccgcatc 4741 gagaagtacg aggacggcgg cgtgctgcac gtgagcttca gctaccgcta cgaggccggc 4801 cgcgtgatcg gcgacttcaa ggtggtgggc accggcttcc ccgaggacag cgtgatcttc 4861 accgacaaga tcatccgcag caacgccacc gtggagcacc tgcaccccat gggcgataac 4921 gtgctggtgg gcagcttcgc ccgcaccttc agcctgcgcg acggcggcta ctacagcttc 4981 gtggtggaca gccacatgca cttcaagagc gccatccacc ccagcatcct gcagaacggg 5041 ggccccatgt tcgccttccg ccgcgtggag gagctgcaca gcaacaccga gctgggcatc 5101 gtggagtacc agcacgcctt caagaccccc atcgccttcg ccagatcccg cgctcagtcg 5161 tccaattctg ccgtggacgg caccgccgga cccggctcca ccggatctcg cgagggcaga 5221 ggaagtcttc taacatgcgg tgacgtggag gagaatcccg gccctatgac cgagtacaag 5281 cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc 5341 gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg 5401 gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg 5461 gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg 5521 gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg 5581 cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg 5641 gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc 5701 cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc 5761 cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc 5821 gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgaaatca acctctggat 5881 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt 5941 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc 6001 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg 6061 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc 6121 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa 6181 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat 6241 tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc 6301 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt 6361 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag 6421 acgagtcgga tctccctttg ggccgcctcc ccgcctggta cctttaagac caatgactta 6481 caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat 6541 tcactcccaa cgaagataag atctgctttt tgcttgtact gggtctctct ggttagacca 6601 gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag 6661 cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag 6721 atccctcaga cccttttagt cagtgtggaa aatctctagc agtagtagtt catgtcatct 6781 tattattcag tatttataac ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt 6841 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc 6901 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt 6961 ctggctctag ctatcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc 7021 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc 7081 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct agacttttgc 7141 agagaccaaa ttcgtaatca tgtcatagct gtttcctgtg tgaaattgtt atccgctcac 7201 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt 7261 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc 7321 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg 7381 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt 7441 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa 7501 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc 7561 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag 7621 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt 7681 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg 7741 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg 7801 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg 7861 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac 7921 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg 7981 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt 8041 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg 8101 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc 8161 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt 8221 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt 8281 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag 8341 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt 8401 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc 8461 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc 8521 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg 8581 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac 8641 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg 8701 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc 8761 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact 8821 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc 8881 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat 8941 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc 9001 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac 9061 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa 9121 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact 9181 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg 9241 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg 9301 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag 9361 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca 9421 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc 9481 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc 9541 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag 9601 gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg 9661 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg 9721 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtgc 9781 caagctg //