LOCUS V038482 9487 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V038482 VERSION V038482 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..3513 /label="SGPP2(NM_152386)" /note="SGPP2(NM_152386)" /gene="SGPP2" CDS 3520..3585 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3629..3840 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3853..4121 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4150..4159 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4180..4842 /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 4912..4965 /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 4966..5565 /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 5566..6154 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6228..6461 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6533..6654 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6694..6829 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6862..6878) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6886..6902 /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(6910..6940) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6955..6976 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7264..7852) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8023..8883) /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(8884..8988) /gene="bla" /label="AmpR promoter" primer_bind 9462..9478 /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 ccgagctgct gcggagcctg 2341 caggattccc agctcgtcgc ccgcttccag cgccgctgcg ggctcttccc cgctccggat 2401 gaaggccccc gggagaacgg cgcggacccc acggagcgcg cggcgcgggt ccccggggtc 2461 gagcatctcc ccgcagccaa cggcaagggc ggcgaggctc cggccaacgg gctgcgcaga 2521 gccgcggcgc cggaggctta tgtacagaag tacgtcgtga agaattattt ctactattac 2581 ctattccaat tttcagctgc tttgggccaa gaagtgttct acatcacgtt tcttccattc 2641 actcactgga atattgaccc ttatttatcc agaagattga tcatcatatg ggttttggtg 2701 atgtatattg gccaagtggc caaggatgtc ttgaagtggc cccgtccctc ctcccctcca 2761 gttgtaaaac tggaaaagag actgatcgct gaatatggaa tgccatccac ccacgccatg 2821 gcggccactg ccattgcctt caccctcctt atctctacta tggacagata ccagtatcca 2881 tttgtgttgg gactggtgat ggccgtggtg ttttccacct tggtgtgtct cagcaggctc 2941 tacactggga tgcatacggt cctggatgtg ctgggtggcg tcctgatcac cgcactcctc 3001 atcgtcctca cctaccctgc ctggaccttc atcgactgcc tggactcggc cagccccctc 3061 ttccccgtgt gtgtcatagt tgtgccattc ttcctgtgtt acaattaccc tgtttctgat 3121 tactacagcc caacccgggc ggacaccacc accattctgg ctgccggggc tggagtgacc 3181 ataggattct ggatcaacca tttcttccag cttgtatcca agcccgctga atctctccct 3241 gttattcaga acatcccacc actcaccacc tacatgttag ttttgggtct gaccaaattt 3301 gcagtgggaa ttgtgttgat cctcttggtt cgtcagcttg tacaaaatct ctcactgcaa 3361 gtattatact catggttcaa ggtggtcacc aggaacaagg aggccaggcg gagactggag 3421 attgaagtgc cttacaagtt tgttacctac acatctgttg gcatctgcgc tacaaccttt 3481 gtgccgatgc ttcacaggtt tctgggatta cccggatccg actacaaaga ccatgacggt 3541 gattataaag atcatgacat cgactacaag gatgacgatg acaagtagcg gccgcgaagg 3601 atctgcgatc gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag 3661 aagttggggg gaggggtcgg caattgaacg ggtgcctaga gaaggtggcg cggggtaaac 3721 tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat 3781 ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag 3841 ctgaagcttc gaggggctcg catctctcct tcacgcgccc gccgccctac ctgaggccgc 3901 catccacgcc ggttgagtcg cgttctgccg cctcccgcct gtggtgcctc ctgaactgcg 3961 tccgccgtct aggtaagttt aaagctcagg tcgagaccgg gcctttgtcc ggcgctccct 4021 tggagcctac ctagactcag ccggctctcc acgctttgcc tgaccctgct tgctcaactc 4081 tacgtctttg tttcgttttc tgttctgcgc cgttacagat ccaagctgtg accggcgcct 4141 acgctagacg ccaccatgga gagcgacgag agcggcctgc ccgccatgga gatcgagtgc 4201 cgcatcaccg gcaccctgaa cggcgtggag ttcgagctgg tgggcggcgg agagggcacc 4261 cccaagcagg gccgcatgac caacaagatg aagagcacca aaggcgccct gaccttcagc 4321 ccctacctgc tgagccacgt gatgggctac ggcttctacc acttcggcac ctaccccagc 4381 ggctacgaga accccttcct gcacgccatc aacaacggcg gctacaccaa cacccgcatc 4441 gagaagtacg aggacggcgg cgtgctgcac gtgagcttca gctaccgcta cgaggccggc 4501 cgcgtgatcg gcgacttcaa ggtggtgggc accggcttcc ccgaggacag cgtgatcttc 4561 accgacaaga tcatccgcag caacgccacc gtggagcacc tgcaccccat gggcgataac 4621 gtgctggtgg gcagcttcgc ccgcaccttc agcctgcgcg acggcggcta ctacagcttc 4681 gtggtggaca gccacatgca cttcaagagc gccatccacc ccagcatcct gcagaacggg 4741 ggccccatgt tcgccttccg ccgcgtggag gagctgcaca gcaacaccga gctgggcatc 4801 gtggagtacc agcacgcctt caagaccccc atcgccttcg ccagatcccg cgctcagtcg 4861 tccaattctg ccgtggacgg caccgccgga cccggctcca ccggatctcg cgagggcaga 4921 ggaagtcttc taacatgcgg tgacgtggag gagaatcccg gccctatgac cgagtacaag 4981 cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc 5041 gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg 5101 gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg 5161 gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg 5221 gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg 5281 cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg 5341 gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc 5401 cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc 5461 cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc 5521 gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgaaatca acctctggat 5581 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt 5641 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc 5701 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg 5761 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc 5821 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa 5881 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat 5941 tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc 6001 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt 6061 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag 6121 acgagtcgga tctccctttg ggccgcctcc ccgcctggta cctttaagac caatgactta 6181 caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat 6241 tcactcccaa cgaagataag atctgctttt tgcttgtact gggtctctct ggttagacca 6301 gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag 6361 cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag 6421 atccctcaga cccttttagt cagtgtggaa aatctctagc agtagtagtt catgtcatct 6481 tattattcag tatttataac ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt 6541 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc 6601 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt 6661 ctggctctag ctatcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc 6721 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc 6781 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct agacttttgc 6841 agagaccaaa ttcgtaatca tgtcatagct gtttcctgtg tgaaattgtt atccgctcac 6901 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt 6961 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc 7021 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg 7081 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt 7141 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa 7201 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc 7261 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag 7321 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt 7381 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg 7441 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg 7501 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg 7561 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac 7621 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg 7681 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt 7741 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg 7801 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc 7861 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt 7921 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt 7981 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag 8041 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt 8101 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc 8161 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc 8221 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg 8281 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac 8341 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg 8401 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc 8461 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact 8521 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc 8581 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat 8641 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc 8701 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac 8761 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa 8821 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact 8881 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg 8941 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg 9001 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag 9061 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca 9121 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc 9181 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc 9241 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag 9301 gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg 9361 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg 9421 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtgc 9481 caagctg //