LOCUS V036255 8950 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V036255 VERSION V036255 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..2976 /label="Dusp19(NM_024438)" /note="Dusp19(NM_024438)" /gene="Dusp19" CDS 2983..3048 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3092..3303 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3316..3584 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 3613..3622 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 3643..4305 /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 4375..4428 /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 4429..5028 /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 5029..5617 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5691..5924 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 5996..6117 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6157..6292 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6325..6341) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6349..6365 /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(6373..6403) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6418..6439 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6727..7315) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7486..8346) /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(8347..8451) /gene="bla" /label="AmpR promoter" primer_bind 8925..8941 /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 gccaccatgc actccctgaa ccaagaaatc 2341 aaagcattct cccgggataa tctcaggaag cagtgcacca gggtgaccac gctaactgga 2401 aagaaactta tagaaacctg ggaagatgcc acagttcatg ttgtggagac agagcccagc 2461 ggtgggggtg gttgtggcta tgtgcaggac cttaccttgg acctgcaagt tggcgttatt 2521 aagccctggt tgcttctggg gtcacaggat gctgctcatg acctggagct actgagaaag 2581 cataaggtga ctcatattct caatgttgca tatggagttg aaaatgcttt cctcagtgag 2641 tttacatata agaccatttc tatactggat gtgcctgaaa ccaatatcct gtcttatttt 2701 ccagaatgtt ttgagtttat tgagcaagca aaactgaagg atggcgtggt tctcgtgcac 2761 tgtaatgcag gtgtttccag ggctgctgca attgtcattg gcttcctcat gagttctgaa 2821 gaagccactt tcaccactgc cctgtcgttg gtgaaagagg cgagaccatc catatgtccg 2881 aatcctggct tcatggaaca actccgcacc taccaagtag gcaaggagag caatggaggt 2941 gacaaagtgc ccgcggagga cacgaccggt ggtctgggat ccgactacaa agaccatgac 3001 ggtgattata aagatcatga catcgactac aaggatgacg atgacaagta gcggccgcga 3061 aggatctgcg atcgctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc 3121 gagaagttgg ggggaggggt cggcaattga acgggtgcct agagaaggtg gcgcggggta 3181 aactgggaaa gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg 3241 tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca 3301 cagctgaagc ttcgaggggc tcgcatctct ccttcacgcg cccgccgccc tacctgaggc 3361 cgccatccac gccggttgag tcgcgttctg ccgcctcccg cctgtggtgc ctcctgaact 3421 gcgtccgccg tctaggtaag tttaaagctc aggtcgagac cgggcctttg tccggcgctc 3481 ccttggagcc tacctagact cagccggctc tccacgcttt gcctgaccct gcttgctcaa 3541 ctctacgtct ttgtttcgtt ttctgttctg cgccgttaca gatccaagct gtgaccggcg 3601 cctacgctag acgccaccat ggagagcgac gagagcggcc tgcccgccat ggagatcgag 3661 tgccgcatca ccggcaccct gaacggcgtg gagttcgagc tggtgggcgg cggagagggc 3721 acccccaagc agggccgcat gaccaacaag atgaagagca ccaaaggcgc cctgaccttc 3781 agcccctacc tgctgagcca cgtgatgggc tacggcttct accacttcgg cacctacccc 3841 agcggctacg agaacccctt cctgcacgcc atcaacaacg gcggctacac caacacccgc 3901 atcgagaagt acgaggacgg cggcgtgctg cacgtgagct tcagctaccg ctacgaggcc 3961 ggccgcgtga tcggcgactt caaggtggtg ggcaccggct tccccgagga cagcgtgatc 4021 ttcaccgaca agatcatccg cagcaacgcc accgtggagc acctgcaccc catgggcgat 4081 aacgtgctgg tgggcagctt cgcccgcacc ttcagcctgc gcgacggcgg ctactacagc 4141 ttcgtggtgg acagccacat gcacttcaag agcgccatcc accccagcat cctgcagaac 4201 gggggcccca tgttcgcctt ccgccgcgtg gaggagctgc acagcaacac cgagctgggc 4261 atcgtggagt accagcacgc cttcaagacc cccatcgcct tcgccagatc ccgcgctcag 4321 tcgtccaatt ctgccgtgga cggcaccgcc ggacccggct ccaccggatc tcgcgagggc 4381 agaggaagtc ttctaacatg cggtgacgtg gaggagaatc ccggccctat gaccgagtac 4441 aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg caccctcgcc 4501 gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg ccacatcgag 4561 cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg 4621 tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg 4681 ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc 4741 gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc 4801 ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg 4861 ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg 4921 ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg 4981 cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgaaa tcaacctctg 5041 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta 5101 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt 5161 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc 5221 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt 5281 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg 5341 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac 5401 aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc 5461 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac 5521 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct 5581 cagacgagtc ggatctccct ttgggccgcc tccccgcctg gtacctttaa gaccaatgac 5641 ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct 5701 aattcactcc caacgaagat aagatctgct ttttgcttgt actgggtctc tctggttaga 5761 ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata 5821 aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta 5881 gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtagta gttcatgtca 5941 tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggaactt 6001 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa 6061 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca 6121 tgtctggctc tagctatccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt 6181 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc 6241 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctagacttt 6301 tgcagagacc aaattcgtaa tcatgtcata gctgtttcct gtgtgaaatt gttatccgct 6361 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg 6421 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct 6481 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg 6541 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc 6601 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg 6661 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct 6721 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca 6781 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct 6841 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc 6901 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt 6961 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc 7021 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc 7081 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg 7141 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc 7201 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag 7261 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga 7321 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat 7381 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag 7441 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat 7501 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc 7561 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat 7621 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag 7681 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg 7741 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc 7801 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca 7861 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg 7921 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc 7981 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta 8041 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc 8101 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg 8161 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc 8221 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc 8281 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat 8341 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag 8401 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc 8461 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa 8521 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg 8581 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca 8641 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc 8701 atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt 8761 aaggagaaaa taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg 8821 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag 8881 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag 8941 tgccaagctg //