LOCUS V038412 9247 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V038412 VERSION V038412 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..3273 /label="ANKRD1(NM_014391)" /note="ANKRD1(NM_014391)" /gene="ANKRD1" CDS 3280..3345 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3389..3600 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3613..3881 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 3910..3919 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 3940..4602 /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 4672..4725 /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 4726..5325 /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 5326..5914 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5988..6221 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6293..6414 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6454..6589 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6622..6638) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6646..6662 /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(6670..6700) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6715..6736 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7024..7612) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7783..8643) /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(8644..8748) /gene="bla" /label="AmpR promoter" primer_bind 9222..9238 /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 gccaccatga tggtactgaa agtagaggaa 2341 ctggtcactg gaaagaagaa tggcaatggg gaggcagggg aattccttcc tgaggatttc 2401 agagatggag agtatgaagc tgctgttact ttagagaagc aggaggatct gaagacactt 2461 ctagcccacc ctgtgaccct gggggagcaa cagtggaaaa gcgagaaaca acgagaggca 2521 gagctcaaaa agaaaaaact agaacaaaga tcaaagcttg aaaatttaga agaccttgaa 2581 ataatcattc aactgaagaa aaggaaaaaa tacaggaaaa ctaaagttcc agttgtaaag 2641 gaaccagaac ctgaaatcat tacggaacct gtggatgtgc ctacgtttct gaaggctgct 2701 ctggagaata aactgccagt agtagaaaaa ttcttgtcag acaagaacaa tccagatgtt 2761 tgtgatgagt ataaacggac agctcttcat agagcatgct tggaaggaca tttggcaatt 2821 gtggagaagt taatggaagc tggagcccag atcgaattcc gtgatatgct tgaatccaca 2881 gccatccact gggcaagccg tggaggaaac ctggatgttt taaaattgtt gctgaataaa 2941 ggagcaaaaa ttagcgcccg agataagttg ctcagcacag cgctgcatgt ggcggtgagg 3001 actggccact atgagtgcgc ggagcatctt atcgcctgtg aggcagacct caacgccaaa 3061 gacagagaag gagatacccc gttgcatgat gcggtgagac tgaaccgcta taagatgatc 3121 cgactcctga ttatgtatgg cgcggatctc aacatcaaga actgtgctgg gaagacgccg 3181 atggatctgg tgctacactg gcagaatgga accaaagcaa tattcgacag cctcagagag 3241 aactcctaca agacctctcg catagctaca ttcggatccg actacaaaga ccatgacggt 3301 gattataaag atcatgacat cgactacaag gatgacgatg acaagtagcg gccgcgaagg 3361 atctgcgatc gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag 3421 aagttggggg gaggggtcgg caattgaacg ggtgcctaga gaaggtggcg cggggtaaac 3481 tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat 3541 ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag 3601 ctgaagcttc gaggggctcg catctctcct tcacgcgccc gccgccctac ctgaggccgc 3661 catccacgcc ggttgagtcg cgttctgccg cctcccgcct gtggtgcctc ctgaactgcg 3721 tccgccgtct aggtaagttt aaagctcagg tcgagaccgg gcctttgtcc ggcgctccct 3781 tggagcctac ctagactcag ccggctctcc acgctttgcc tgaccctgct tgctcaactc 3841 tacgtctttg tttcgttttc tgttctgcgc cgttacagat ccaagctgtg accggcgcct 3901 acgctagacg ccaccatgga gagcgacgag agcggcctgc ccgccatgga gatcgagtgc 3961 cgcatcaccg gcaccctgaa cggcgtggag ttcgagctgg tgggcggcgg agagggcacc 4021 cccaagcagg gccgcatgac caacaagatg aagagcacca aaggcgccct gaccttcagc 4081 ccctacctgc tgagccacgt gatgggctac ggcttctacc acttcggcac ctaccccagc 4141 ggctacgaga accccttcct gcacgccatc aacaacggcg gctacaccaa cacccgcatc 4201 gagaagtacg aggacggcgg cgtgctgcac gtgagcttca gctaccgcta cgaggccggc 4261 cgcgtgatcg gcgacttcaa ggtggtgggc accggcttcc ccgaggacag cgtgatcttc 4321 accgacaaga tcatccgcag caacgccacc gtggagcacc tgcaccccat gggcgataac 4381 gtgctggtgg gcagcttcgc ccgcaccttc agcctgcgcg acggcggcta ctacagcttc 4441 gtggtggaca gccacatgca cttcaagagc gccatccacc ccagcatcct gcagaacggg 4501 ggccccatgt tcgccttccg ccgcgtggag gagctgcaca gcaacaccga gctgggcatc 4561 gtggagtacc agcacgcctt caagaccccc atcgccttcg ccagatcccg cgctcagtcg 4621 tccaattctg ccgtggacgg caccgccgga cccggctcca ccggatctcg cgagggcaga 4681 ggaagtcttc taacatgcgg tgacgtggag gagaatcccg gccctatgac cgagtacaag 4741 cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc 4801 gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg 4861 gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg 4921 gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg 4981 gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg 5041 cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg 5101 gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc 5161 cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc 5221 cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc 5281 gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgaaatca acctctggat 5341 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt 5401 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc 5461 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg 5521 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc 5581 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa 5641 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat 5701 tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc 5761 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt 5821 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag 5881 acgagtcgga tctccctttg ggccgcctcc ccgcctggta cctttaagac caatgactta 5941 caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat 6001 tcactcccaa cgaagataag atctgctttt tgcttgtact gggtctctct ggttagacca 6061 gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag 6121 cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag 6181 atccctcaga cccttttagt cagtgtggaa aatctctagc agtagtagtt catgtcatct 6241 tattattcag tatttataac ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt 6301 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc 6361 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt 6421 ctggctctag ctatcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc 6481 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc 6541 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct agacttttgc 6601 agagaccaaa ttcgtaatca tgtcatagct gtttcctgtg tgaaattgtt atccgctcac 6661 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt 6721 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc 6781 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg 6841 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt 6901 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa 6961 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc 7021 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag 7081 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt 7141 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg 7201 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg 7261 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg 7321 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac 7381 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg 7441 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt 7501 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg 7561 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc 7621 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt 7681 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt 7741 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag 7801 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt 7861 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc 7921 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc 7981 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg 8041 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac 8101 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg 8161 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc 8221 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact 8281 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc 8341 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat 8401 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc 8461 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac 8521 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa 8581 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact 8641 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg 8701 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg 8761 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag 8821 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca 8881 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc 8941 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc 9001 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag 9061 gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg 9121 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg 9181 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtgc 9241 caagctg //