LOCUS V036077 9730 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V036077 VERSION V036077 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..3756 /label="EDIL3(NM_005711)" /note="EDIL3(NM_005711)" /gene="EDIL3" CDS 3763..3828 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3872..4083 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4096..4364 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4393..4402 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4423..5085 /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 5155..5208 /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 5209..5808 /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 5809..6397 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6471..6704 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6776..6897 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6937..7072 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(7105..7121) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7129..7145 /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(7153..7183) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7198..7219 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7507..8095) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8266..9126) /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(9127..9231) /gene="bla" /label="AmpR promoter" primer_bind 9705..9721 /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 agcgctcggt agccgtctgg 2341 ctcttggtcg ggctcagcct cggtgtcccc cagttcggca aaggtgatat ttgtgatccc 2401 aatccatgtg aaaatggagg tatctgtttg ccaggattgg ctgatggttc cttttcctgt 2461 gagtgtccag atggcttcac agaccccaac tgttctagtg ttgtggaggt tgcatcagat 2521 gaagaagaac caacttcagc aggtccctgc actcctaatc catgccataa tggaggaacc 2581 tgtgaaataa gtgaagcata ccgaggggat acattcatag gctatgtttg taaatgtccc 2641 cgaggattta atgggattca ctgtcagcac aacataaatg aatgcgaagt tgagccttgc 2701 aaaaatggtg gaatatgtac agatcttgtt gctaactatt cctgtgagtg cccaggcgaa 2761 tttatgggaa gaaattgtca atacaaatgc tcaggcccac tgggaattga aggtggaatt 2821 atatcaaacc agcaaatcac agcttcctct actcaccgag ctctttttgg actccaaaaa 2881 tggtatccct actatgcacg tcttaataag aaggggctta taaatgcgtg gacagctgca 2941 gaaaatgaca gatggccgtg gattcagata aatttgcaaa ggaaaatgag agttactggt 3001 gtgattaccc aaggagccaa gaggattgga agcccagagt atataaaatc ctacaaaatt 3061 gcctacagta atgatggaaa gacttgggca atgtacaaag tgaaaggcac caatgaagac 3121 atggtgtttc gtggaaacat tgataacaac actccatatg ctaactcttt cacacccccc 3181 ataaaagctc agtatgtaag actctatccc caagtttgtc gaagacattg cactttgcga 3241 atggaacttc ttggctgtga actgtcgggt tgttctgagc ctctgggtat gaaatcagga 3301 catatacaag actatcagat cactgcctcc agcatcttca gaacgctcaa catggacatg 3361 ttcacttggg aaccaaggaa agctcggctg gacaagcaag gcaaagtgaa tgcctggacc 3421 tctggccaca atgaccagtc acaatggtta caggtggatc ttcttgttcc aaccaaagtg 3481 actggcatca ttacacaagg agctaaagat tttggtcatg tacagtttgt tggctcctac 3541 aaactggctt acagcaatga tggagaacac tggactgtat accaggatga aaagcaaaga 3601 aaagataagg ttttccaggg aaattttgac aatgacactc acagaaaaaa tgtcatcgac 3661 cctcccatct atgcacgaca cataagaatc cttccttggt cctggtacgg gaggatcaca 3721 ttgcggtcag agctgctggg ctgcacagag gaggaaggat ccgactacaa agaccatgac 3781 ggtgattata aagatcatga catcgactac aaggatgacg atgacaagta gcggccgcga 3841 aggatctgcg atcgctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc 3901 gagaagttgg ggggaggggt cggcaattga acgggtgcct agagaaggtg gcgcggggta 3961 aactgggaaa gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg 4021 tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca 4081 cagctgaagc ttcgaggggc tcgcatctct ccttcacgcg cccgccgccc tacctgaggc 4141 cgccatccac gccggttgag tcgcgttctg ccgcctcccg cctgtggtgc ctcctgaact 4201 gcgtccgccg tctaggtaag tttaaagctc aggtcgagac cgggcctttg tccggcgctc 4261 ccttggagcc tacctagact cagccggctc tccacgcttt gcctgaccct gcttgctcaa 4321 ctctacgtct ttgtttcgtt ttctgttctg cgccgttaca gatccaagct gtgaccggcg 4381 cctacgctag acgccaccat ggagagcgac gagagcggcc tgcccgccat ggagatcgag 4441 tgccgcatca ccggcaccct gaacggcgtg gagttcgagc tggtgggcgg cggagagggc 4501 acccccaagc agggccgcat gaccaacaag atgaagagca ccaaaggcgc cctgaccttc 4561 agcccctacc tgctgagcca cgtgatgggc tacggcttct accacttcgg cacctacccc 4621 agcggctacg agaacccctt cctgcacgcc atcaacaacg gcggctacac caacacccgc 4681 atcgagaagt acgaggacgg cggcgtgctg cacgtgagct tcagctaccg ctacgaggcc 4741 ggccgcgtga tcggcgactt caaggtggtg ggcaccggct tccccgagga cagcgtgatc 4801 ttcaccgaca agatcatccg cagcaacgcc accgtggagc acctgcaccc catgggcgat 4861 aacgtgctgg tgggcagctt cgcccgcacc ttcagcctgc gcgacggcgg ctactacagc 4921 ttcgtggtgg acagccacat gcacttcaag agcgccatcc accccagcat cctgcagaac 4981 gggggcccca tgttcgcctt ccgccgcgtg gaggagctgc acagcaacac cgagctgggc 5041 atcgtggagt accagcacgc cttcaagacc cccatcgcct tcgccagatc ccgcgctcag 5101 tcgtccaatt ctgccgtgga cggcaccgcc ggacccggct ccaccggatc tcgcgagggc 5161 agaggaagtc ttctaacatg cggtgacgtg gaggagaatc ccggccctat gaccgagtac 5221 aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg caccctcgcc 5281 gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg ccacatcgag 5341 cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg 5401 tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg 5461 ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc 5521 gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc 5581 ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg 5641 ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg 5701 ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg 5761 cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgaaa tcaacctctg 5821 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta 5881 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt 5941 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc 6001 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt 6061 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg 6121 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac 6181 aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc 6241 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac 6301 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct 6361 cagacgagtc ggatctccct ttgggccgcc tccccgcctg gtacctttaa gaccaatgac 6421 ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct 6481 aattcactcc caacgaagat aagatctgct ttttgcttgt actgggtctc tctggttaga 6541 ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata 6601 aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta 6661 gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtagta gttcatgtca 6721 tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggaactt 6781 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa 6841 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca 6901 tgtctggctc tagctatccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt 6961 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc 7021 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctagacttt 7081 tgcagagacc aaattcgtaa tcatgtcata gctgtttcct gtgtgaaatt gttatccgct 7141 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg 7201 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct 7261 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg 7321 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc 7381 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg 7441 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct 7501 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca 7561 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct 7621 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc 7681 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt 7741 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc 7801 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc 7861 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg 7921 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc 7981 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag 8041 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga 8101 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat 8161 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag 8221 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat 8281 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc 8341 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat 8401 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag 8461 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg 8521 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc 8581 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca 8641 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg 8701 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc 8761 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta 8821 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc 8881 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg 8941 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc 9001 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc 9061 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat 9121 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag 9181 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc 9241 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa 9301 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg 9361 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca 9421 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc 9481 atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt 9541 aaggagaaaa taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg 9601 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag 9661 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag 9721 tgccaagctg //