LOCUS V039981 10207 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V039981 VERSION V039981 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..4233 /label="ILDR2(NM_199351)" /note="ILDR2(NM_199351)" /gene="ILDR2" CDS 4240..4305 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 4349..4560 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4573..4841 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4870..4879 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4900..5562 /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 5632..5685 /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 5686..6285 /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 6286..6874 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6948..7181 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 7253..7374 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 7414..7549 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(7582..7598) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7606..7622 /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(7630..7660) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7675..7696 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7984..8572) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8743..9603) /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(9604..9708) /gene="bla" /label="AmpR promoter" primer_bind 10182..10198 /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 atagggtctt gctgaggtgg 2341 atttctctct tctggctaac agccatggtc gaaggccttc aggtcacagt gcccgacaag 2401 aagaaggtgg ccatgctctt ccagcccact gtgcttcgct gccacttctc aacatcctcc 2461 catcagcctg cagttgtgca gtggaagttc aagtcctact gccaggatcg catgggagaa 2521 tccttgggca tgtcctctac ccgggcccaa tctctcagca agagaaacct ggaatgggac 2581 ccctacttgg attgtttgga cagcaggagg actgttcgag tagtagcttc aaaacagggc 2641 tcgactgtca ccctgggaga tttctacagg ggcagagaga tcacgattgt tcatgatgca 2701 gatcttcaaa ttggaaagct tatgtgggga gacagcggac tctattactg tattatcacc 2761 accccagatg acctggaggg gaaaaatgag gactcagtgg aactgctggt gttgggcagg 2821 acagggctgc ttgctgatct cttgcccagt tttgctgtgg agattatgcc agagtgggtg 2881 tttgttggcc tggtgctcct gggcgtcttc ctcttcttcg tcctggtggg gatctgctgg 2941 tgccagtgct gccctcacag ctgctgctgc tatgtccgct gcccatgctg cccagattcc 3001 tgctgctgcc ctcaagcctt gtatgaagca gggaaagcag caaaggccgg gtaccctccc 3061 tctgtctccg gtgtccccgg cccttactcc atcccctctg tccctttggg aggagccccc 3121 tcatctggca tgctgatgga caagccgcat ccacctccct tggcaccaag tgactccact 3181 ggaggaagcc acagtgttcg caaaggttac cggatccagg ctgacaaaga gagagactcc 3241 atgaaggtcc tgtactatgt tgagaaggag ctggctcagt ttgatccagc cagaaggatg 3301 agaggcagat ataacaacac catctcagaa ctcagctccc tacatgagga ggacagcaat 3361 ttccgccagt ctttccatca gatgagaagc aagcagttcc ctgtgtctgg ggacttggag 3421 agcaatcctg actattggtc aggtgtcatg ggaggcagca gtggggcaag ccgcgggccc 3481 tcagccatgg agtataacaa agaggatcga gagagcttca ggcacagcca gccgcgctcc 3541 aagtcggaga tgctgtcgcg gaagaacttc gccacggggg tgccggccgt ttccatggac 3601 gagctggcgg ccttcgctga ctcctacggc cagcggcccc gccgggcaga cggcaacagt 3661 cacgaggcgc ggggcgggag ccgcttcgag cgctcggagt cgcgggcgca cagcggcttc 3721 taccaggacg actccttgga ggagtactac ggtcagcgca gccgcagccg cgagcccctg 3781 accgatgctg accgcggctg ggccttcagc cccgcgcgcc gcagacccgc cgaggacgcg 3841 cacctgccgc ggctggtgag ccgcacgcca ggcaccgcac ccaaatacga ccactcgtac 3901 ctgggcagcg cgcgggagcg ccaggcgcgg cccgagggcg ccagccgcgg tggcagcctg 3961 gagacgccat ccaagcggag cgcgcagctc ggcccgcgca gcgcctccta ctacgcttgg 4021 tcgccgcccg gcacctacaa ggccggctcg tcgcaggacg accaggagga cgcgtccgac 4081 gacgcgctgc cgccctacag cgagctggag ctgacccgcg gcccgtccta ccgcggccgc 4141 gacctgccct accacagcaa ctcggagaag aagaggaaaa aggagcccgc caagaaaacc 4201 aatgactttc caaccaggat gtcccttgtg gtcggatccg actacaaaga ccatgacggt 4261 gattataaag atcatgacat cgactacaag gatgacgatg acaagtagcg gccgcgaagg 4321 atctgcgatc gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag 4381 aagttggggg gaggggtcgg caattgaacg ggtgcctaga gaaggtggcg cggggtaaac 4441 tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat 4501 ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag 4561 ctgaagcttc gaggggctcg catctctcct tcacgcgccc gccgccctac ctgaggccgc 4621 catccacgcc ggttgagtcg cgttctgccg cctcccgcct gtggtgcctc ctgaactgcg 4681 tccgccgtct aggtaagttt aaagctcagg tcgagaccgg gcctttgtcc ggcgctccct 4741 tggagcctac ctagactcag ccggctctcc acgctttgcc tgaccctgct tgctcaactc 4801 tacgtctttg tttcgttttc tgttctgcgc cgttacagat ccaagctgtg accggcgcct 4861 acgctagacg ccaccatgga gagcgacgag agcggcctgc ccgccatgga gatcgagtgc 4921 cgcatcaccg gcaccctgaa cggcgtggag ttcgagctgg tgggcggcgg agagggcacc 4981 cccaagcagg gccgcatgac caacaagatg aagagcacca aaggcgccct gaccttcagc 5041 ccctacctgc tgagccacgt gatgggctac ggcttctacc acttcggcac ctaccccagc 5101 ggctacgaga accccttcct gcacgccatc aacaacggcg gctacaccaa cacccgcatc 5161 gagaagtacg aggacggcgg cgtgctgcac gtgagcttca gctaccgcta cgaggccggc 5221 cgcgtgatcg gcgacttcaa ggtggtgggc accggcttcc ccgaggacag cgtgatcttc 5281 accgacaaga tcatccgcag caacgccacc gtggagcacc tgcaccccat gggcgataac 5341 gtgctggtgg gcagcttcgc ccgcaccttc agcctgcgcg acggcggcta ctacagcttc 5401 gtggtggaca gccacatgca cttcaagagc gccatccacc ccagcatcct gcagaacggg 5461 ggccccatgt tcgccttccg ccgcgtggag gagctgcaca gcaacaccga gctgggcatc 5521 gtggagtacc agcacgcctt caagaccccc atcgccttcg ccagatcccg cgctcagtcg 5581 tccaattctg ccgtggacgg caccgccgga cccggctcca ccggatctcg cgagggcaga 5641 ggaagtcttc taacatgcgg tgacgtggag gagaatcccg gccctatgac cgagtacaag 5701 cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc 5761 gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg 5821 gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg 5881 gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg 5941 gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg 6001 cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg 6061 gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc 6121 cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc 6181 cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc 6241 gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgaaatca acctctggat 6301 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt 6361 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc 6421 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg 6481 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc 6541 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa 6601 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat 6661 tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc 6721 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt 6781 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag 6841 acgagtcgga tctccctttg ggccgcctcc ccgcctggta cctttaagac caatgactta 6901 caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat 6961 tcactcccaa cgaagataag atctgctttt tgcttgtact gggtctctct ggttagacca 7021 gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag 7081 cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag 7141 atccctcaga cccttttagt cagtgtggaa aatctctagc agtagtagtt catgtcatct 7201 tattattcag tatttataac ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt 7261 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc 7321 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt 7381 ctggctctag ctatcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc 7441 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc 7501 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct agacttttgc 7561 agagaccaaa ttcgtaatca tgtcatagct gtttcctgtg tgaaattgtt atccgctcac 7621 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt 7681 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc 7741 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg 7801 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt 7861 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa 7921 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc 7981 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag 8041 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt 8101 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg 8161 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg 8221 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg 8281 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac 8341 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg 8401 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt 8461 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg 8521 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc 8581 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt 8641 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt 8701 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag 8761 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt 8821 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc 8881 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc 8941 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg 9001 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac 9061 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg 9121 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc 9181 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact 9241 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc 9301 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat 9361 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc 9421 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac 9481 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa 9541 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact 9601 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg 9661 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg 9721 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag 9781 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca 9841 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc 9901 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc 9961 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag 10021 gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg 10081 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg 10141 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtgc 10201 caagctg //