LOCUS V038398 9985 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V038398 VERSION V038398 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..4011 /label="USP39(NM_006590)" /note="USP39(NM_006590)" /gene="USP39" CDS 4018..4083 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 4127..4338 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4351..4619 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4648..4657 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4678..5340 /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 5410..5463 /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 5464..6063 /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 6064..6652 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6726..6959 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 7031..7152 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 7192..7327 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(7360..7376) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7384..7400 /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(7408..7438) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7453..7474 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7762..8350) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8521..9381) /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(9382..9486) /gene="bla" /label="AmpR promoter" primer_bind 9960..9976 /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 gccaccatgt ccggccggtc taagcgggag 2341 tctcgcggtt ccactcgcgg gaagcgagag tctgagtcgc ggggcagctc cggtcgcgtc 2401 aagcgggagc gagatcggga gcgggagcct gaggcggcga gctcccgggg cagccctgtg 2461 cgcgtgaagc gggagttcga gccggcgagc gcgcgcgagg ccccggcttc tgttgtcccg 2521 tttgtgcggg tgaagcggga gcgcgaggtc gatgaggact cggagcctga gcgggaggtg 2581 cgagcaaaga atggccgagt ggattctgag gaccggagga gccgccactg cccgtacctg 2641 gacaccatta acaggagtgt gctggacttt gactttgaga aactgtgttc tatctccctc 2701 tcacacatca atgcttatgc ctgtctggtg tgtggcaagt actttcaagg ccggggtttg 2761 aagtctcacg cctacattca cagtgtccag tttagccacc atgttttcct caacctccac 2821 accctcaagt tttactgcct tccagacaac tatgagatca tcgattcctc attggaggat 2881 atcacgtatg tgttgaagcc cactttcaca aagcagcaaa ttgcaaactt ggacaagcaa 2941 gccaaattgt cccgggcata tgatggtacc acttacctgc cgggtattgt gggactgaat 3001 aacataaagg ccaatgatta tgccaacgct gtccttcagg ctctatctaa tgttcctcct 3061 ctccggaact actttctgga agaagacaat tataagaaca tcaaacgtcc tccaggggat 3121 atcatgttct tgttggtcca gcgttttgga gagctgatga gaaagctctg gaaccctcga 3181 aatttcaagg cacatgtgtc tccccatgag atgcttcagg cagttgtact ttgcagtaag 3241 aagacttttc agatcaccaa acaaggagat ggcgttgact ttctgtcttg gtttctgaat 3301 gctctgcact cagctctggg gggcacaaag aagaaaaaga agactattgt gactgatgtt 3361 ttccaggggt ccatgaggat cttcactaaa aagcttcccc atcctgatct gccagcagaa 3421 gaaaaagagc agttgctcca taatgacgag taccaggaga caatggtgga gtccactttt 3481 atgtacctga cgctggacct tcctactgcc cccctctaca aggacgagaa ggagcagctc 3541 atcattcccc aagtgccact cttcaacatc ctggctaagt tcaatggcat cactgagaag 3601 gaatataaga cttacaagga gaactttctg aagcgcttcc agcttaccaa gttgcctcca 3661 tatctaatct tttgtatcaa gagattcact aagaacaact tctttgttga gaagaatcca 3721 actattgtca atttccctat tacaaatgtg gatctgagag aatacttgtc tgaagaagta 3781 caagcagtac acaagaatac cacctatgac ctcattgcca acatcgtgca tgacggcaag 3841 ccctccgagg gctcctaccg gatccacgtg cttcatcatg ggacaggcaa atggtatgaa 3901 ttacaagacc tccaggtgac tgacatcctt ccccagatga tcacactgtc agaggcttac 3961 attcagattt ggaagaggcg agataatgat gaaaccaacc agcagggggc tggatccgac 4021 tacaaagacc atgacggtga ttataaagat catgacatcg actacaagga tgacgatgac 4081 aagtagcggc cgcgaaggat ctgcgatcgc tccggtgccc gtcagtgggc agagcgcaca 4141 tcgcccacag tccccgagaa gttgggggga ggggtcggca attgaacggg tgcctagaga 4201 aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag 4261 ggtgggggag aaccgtatat aagtgcagta gtcgccgtga acgttctttt tcgcaacggg 4321 tttgccgcca gaacacagct gaagcttcga ggggctcgca tctctccttc acgcgcccgc 4381 cgccctacct gaggccgcca tccacgccgg ttgagtcgcg ttctgccgcc tcccgcctgt 4441 ggtgcctcct gaactgcgtc cgccgtctag gtaagtttaa agctcaggtc gagaccgggc 4501 ctttgtccgg cgctcccttg gagcctacct agactcagcc ggctctccac gctttgcctg 4561 accctgcttg ctcaactcta cgtctttgtt tcgttttctg ttctgcgccg ttacagatcc 4621 aagctgtgac cggcgcctac gctagacgcc accatggaga gcgacgagag cggcctgccc 4681 gccatggaga tcgagtgccg catcaccggc accctgaacg gcgtggagtt cgagctggtg 4741 ggcggcggag agggcacccc caagcagggc cgcatgacca acaagatgaa gagcaccaaa 4801 ggcgccctga ccttcagccc ctacctgctg agccacgtga tgggctacgg cttctaccac 4861 ttcggcacct accccagcgg ctacgagaac cccttcctgc acgccatcaa caacggcggc 4921 tacaccaaca cccgcatcga gaagtacgag gacggcggcg tgctgcacgt gagcttcagc 4981 taccgctacg aggccggccg cgtgatcggc gacttcaagg tggtgggcac cggcttcccc 5041 gaggacagcg tgatcttcac cgacaagatc atccgcagca acgccaccgt ggagcacctg 5101 caccccatgg gcgataacgt gctggtgggc agcttcgccc gcaccttcag cctgcgcgac 5161 ggcggctact acagcttcgt ggtggacagc cacatgcact tcaagagcgc catccacccc 5221 agcatcctgc agaacggggg ccccatgttc gccttccgcc gcgtggagga gctgcacagc 5281 aacaccgagc tgggcatcgt ggagtaccag cacgccttca agacccccat cgccttcgcc 5341 agatcccgcg ctcagtcgtc caattctgcc gtggacggca ccgccggacc cggctccacc 5401 ggatctcgcg agggcagagg aagtcttcta acatgcggtg acgtggagga gaatcccggc 5461 cctatgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc 5521 gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgatccg 5581 gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc 5641 gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg 5701 gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc 5761 ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag 5821 gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg 5881 ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc 5941 ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc 6001 gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc 6061 tgaaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt 6121 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc 6181 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag 6241 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc 6301 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc 6361 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg 6421 ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg 6481 ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc 6541 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt 6601 cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc gcctggtacc 6661 tttaagacca atgacttaca aggcagctgt agatcttagc cactttttaa aagaaaaggg 6721 gggactggaa gggctaattc actcccaacg aagataagat ctgctttttg cttgtactgg 6781 gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact 6841 gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg 6901 tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag 6961 tagtagttca tgtcatctta ttattcagta tttataactt gcaaagaaat gaatatcaga 7021 gagtgagagg aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac 7081 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat 7141 caatgtatct tatcatgtct ggctctagct atcccgcccc taactccgcc catcccgccc 7201 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat 7261 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt 7321 ggaggcctag acttttgcag agaccaaatt cgtaatcatg tcatagctgt ttcctgtgtg 7381 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc 7441 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt 7501 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg 7561 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt 7621 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc 7681 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa 7741 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa 7801 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc 7861 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc 7921 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag 7981 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga 8041 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc 8101 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac 8161 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg 8221 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca 8281 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa 8341 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa 8401 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt 8461 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag 8521 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat 8581 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc 8641 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa 8701 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca 8761 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa 8821 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt 8881 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc 8941 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact 9001 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc 9061 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg 9121 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct 9181 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc 9241 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag 9301 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac 9361 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg 9421 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt 9481 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac 9541 attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga 9601 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga 9661 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg 9721 gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat 9781 accgcacaga tgcgtaagga gaaaataccg catcaggcgc cattcgccat tcaggctgcg 9841 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg 9901 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg 9961 taaaacgacg gccagtgcca agctg //