LOCUS V032046 9207 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V032046 VERSION V032046 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 2293..3288 /label="Ldha(NM_010699)" /note="Ldha(NM_010699)" /gene="Ldha" CDS 3292..3321 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /transl_table=1 /translation="EQKLISEEDL" /label="Myc" promoter 3366..3577 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3590..3858 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 3887..3896 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 3917..4579 /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 4649..4702 /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 4703..5302 /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 5303..5890 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5964..6197 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6269..6390 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6409..6544 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6582..6598) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6606..6622 /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(6630..6660) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6675..6696 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6984..7572) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7743..8603) /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(8604..8708) /gene="bla" /label="AmpR promoter" primer_bind 9182..9198 /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 gaatggcaac cctcaaggac cagctgattg tgaatcttct taaggaagag 2341 caggctcccc agaacaagat tacagttgtt ggggttggtg ctgttggcat ggcttgtgcc 2401 atcagtatct taatgaagga cttggcggat gagcttgccc ttgttgacgt catggaagac 2461 aaactcaagg gcgagatgat ggatctccag catggcagcc tcttccttaa aacaccaaaa 2521 attgtctcca gcaaagacta ctgtgtaact gcgaactcca agctggtcat tatcaccgcg 2581 ggggcccgtc agcaagaggg ggagagccgg ctcaacctgg tccagcgaaa cgtgaacatc 2641 ttcaagttca tcattcccaa cattgtcaag tacagtccac actgcaagct gctgatcgtc 2701 tccaatccag tggatatctt gacctacgtg gcttggaaaa tcagtggctt tcccaaaaac 2761 cgagtaattg gaagtggttg caatctggat tcagcgcggt tccgttacct gatgggagag 2821 aggctggggg ttcacgcgct gagctgtcac ggctgggtcc tgggagaaca tggcgactcc 2881 agtgtgcctg tgtggagtgg tgtgaatgtt gccggcgtct ccctgaagtc tcttaaccca 2941 gaactgggca ctgacgcaga caaggagcag tggaaggagg ttcacaagca ggtggtggac 3001 agtgcctacg aggtgatcaa gctgaaaggt tacacatcct gggccattgg cctctctgtg 3061 gcagacttgg ctgagagcat aatgaagaac cttaggcggg tgcatcccat ttccaccatg 3121 attaagggtc tctatggaat caatgaggat gtcttcctca gtgtcccatg tatcctggga 3181 caaaatggaa tctcggatgt tgtgaaggtg acactgactc ctgaggaaga ggcccgcctg 3241 aagaagagcg cagacaccct ctggggaatc cagaaggagc tgcagttctc cgaacaaaaa 3301 ctcatctcag aagaggatct gtgagcggcc gcgaaggatc tgcgatcgct ccggtgcccg 3361 tcagtgggca gagcgcacat cgcccacagt ccccgagaag ttggggggag gggtcggcaa 3421 ttgaacgggt gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg 3481 gctccgcctt tttcccgagg gtgggggaga accgtatata agtgcagtag tcgccgtgaa 3541 cgttcttttt cgcaacgggt ttgccgccag aacacagctg aagcttcgag gggctcgcat 3601 ctctccttca cgcgcccgcc gccctacctg aggccgccat ccacgccggt tgagtcgcgt 3661 tctgccgcct cccgcctgtg gtgcctcctg aactgcgtcc gccgtctagg taagtttaaa 3721 gctcaggtcg agaccgggcc tttgtccggc gctcccttgg agcctaccta gactcagccg 3781 gctctccacg ctttgcctga ccctgcttgc tcaactctac gtctttgttt cgttttctgt 3841 tctgcgccgt tacagatcca agctgtgacc ggcgcctacg ctagacgcca ccatggagag 3901 cgacgagagc ggcctgcccg ccatggagat cgagtgccgc atcaccggca ccctgaacgg 3961 cgtggagttc gagctggtgg gcggcggaga gggcaccccc aagcagggcc gcatgaccaa 4021 caagatgaag agcaccaaag gcgccctgac cttcagcccc tacctgctga gccacgtgat 4081 gggctacggc ttctaccact tcggcaccta ccccagcggc tacgagaacc ccttcctgca 4141 cgccatcaac aacggcggct acaccaacac ccgcatcgag aagtacgagg acggcggcgt 4201 gctgcacgtg agcttcagct accgctacga ggccggccgc gtgatcggcg acttcaaggt 4261 ggtgggcacc ggcttccccg aggacagcgt gatcttcacc gacaagatca tccgcagcaa 4321 cgccaccgtg gagcacctgc accccatggg cgataacgtg ctggtgggca gcttcgcccg 4381 caccttcagc ctgcgcgacg gcggctacta cagcttcgtg gtggacagcc acatgcactt 4441 caagagcgcc atccacccca gcatcctgca gaacgggggc cccatgttcg ccttccgccg 4501 cgtggaggag ctgcacagca acaccgagct gggcatcgtg gagtaccagc acgccttcaa 4561 gacccccatc gccttcgcca gatcccgcgc tcagtcgtcc aattctgccg tggacggcac 4621 cgccggaccc ggctccaccg gatctcgcga gggcagagga agtcttctaa catgcggtga 4681 cgtggaggag aatcccggcc ctatgaccga gtacaagccc acggtgcgcc tcgccacccg 4741 cgacgacgtc cccagggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac 4801 gcgccacacc gtcgatccgg accgccacat cgagcgggtc accgagctgc aagaactctt 4861 cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt 4921 ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc 4981 gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct 5041 ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga 5101 ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg 5161 cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg 5221 gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat 5281 gacccgcaag cccggtgcct gaaatcaacc tctggattac aaaatttgtg aaagattgac 5341 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt 5401 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt 5461 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt 5521 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg 5581 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg 5641 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaagct 5701 gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt 5761 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc 5821 tctgcggcct cttccgcgtc tccgccttcg ccctcagacg agtcggatct ccctttggcc 5881 gcctccccgc ctggtacctt taagaccaat gacttacaag gcagctgtag atcttagcca 5941 ctttttaaaa gaaaaggggg gactggaagg gctaattcac tcccaacgaa aataagatct 6001 gctttttgct tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg 6061 ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt 6121 gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt 6181 gtggaaaatc tctagcagta gtagttcatg tcatcttatt attcagtatt tataacttgc 6241 aaagaaatga atatcagaga gtgagaggaa cttgtttatt gcagcttata atggttacaa 6301 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg 6361 tggtttgtcc aaactcatca atgtatctta tcatgtctgg ctctagctat cccgccccta 6421 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca 6481 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga 6541 ggcctagact tttgcagaga cggcccaaat tcgtaatcat ggtcatagct gtttcctgtg 6601 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa 6661 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct 6721 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga 6781 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc 6841 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa 6901 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt 6961 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa 7021 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt 7081 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg 7141 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc 7201 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc 7261 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta 7321 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct 7381 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc 7441 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa 7501 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa 7561 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa 7621 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt 7681 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac 7741 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc 7801 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc 7861 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata 7921 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc 7981 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc 8041 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca 8101 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa 8161 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca 8221 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt 8281 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt 8341 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg 8401 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga 8461 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc 8521 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg 8581 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag 8641 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg 8701 gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg 8761 acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat 8821 gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg 8881 gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc 8941 tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa 9001 ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc attcaggctg 9061 cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa 9121 gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt 9181 tgtaaaacga cggccagtgc caagctg //