LOCUS V026152 9070 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V026152 VERSION V026152 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..3096 /label="Mblac1(NM_177878)" /note="Mblac1(NM_177878)" /gene="Mblac1" CDS 3103..3168 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3212..3423 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3436..3704 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 3733..3742 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 3763..4425 /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 4495..4548 /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 4549..5148 /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 5149..5737 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5811..6044 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6116..6237 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6277..6412 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6445..6461) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6469..6485 /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(6493..6523) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6538..6559 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6847..7435) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7606..8466) /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(8467..8571) /gene="bla" /label="AmpR promoter" primer_bind 9045..9061 /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 acggtccagt gcgcaccgag 2341 cccctgcatg gtgagatccc tttgctggcg tccagcggct cctactccgt ggtggttctg 2401 ctgcgcggct acgcggagcc gcagggagcg ggcgacgcgg tgcgtgctga tggcactgtg 2461 actctcgtcc tgcctcgggg ctgggcctcg gactccagcc gcggtttggc cccatccgca 2521 gacggcggct ctaagactgc cctggaggag gcggtgcgtg gccccatcct cgtggacacc 2581 gggggaccct gggcgcgggg tgcgctgctg gaagccctgg ctacacaggg cgtggccccc 2641 gaagatgtga ctctggtggt gggcacccac ggacactcgg atcatattgg gaacctgggg 2701 ctgtttcccg aggcagcttt gttagtctcc catgacttct gccttcccga gggtctctat 2761 ctgccccatg ggctgtgtga aacgcagccc ttgatactgg gctcgggact gcaggtgtgg 2821 gccacgccgg gccacggagg acagcgggac gtgagcgtcg tagtggaggg caccagcctg 2881 ggcaccgtgg tggtggctgg cgatgtgttc gagcgacttg gtgacgagga ctcctggcag 2941 gccctgagcg aagacccagt agctcagcag cggagccggg agaggattct gagcgtagct 3001 gatgtggtgg tgcctggtca cggagccccc tttcgagtgg tcagggaaac agtgaaaagt 3061 tcagaggatt tgatatgtga aggcaaggcg gttgctggat ccgactacaa agaccatgac 3121 ggtgattata aagatcatga catcgactac aaggatgacg atgacaagta gcggccgcga 3181 aggatctgcg atcgctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc 3241 gagaagttgg ggggaggggt cggcaattga acgggtgcct agagaaggtg gcgcggggta 3301 aactgggaaa gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg 3361 tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca 3421 cagctgaagc ttcgaggggc tcgcatctct ccttcacgcg cccgccgccc tacctgaggc 3481 cgccatccac gccggttgag tcgcgttctg ccgcctcccg cctgtggtgc ctcctgaact 3541 gcgtccgccg tctaggtaag tttaaagctc aggtcgagac cgggcctttg tccggcgctc 3601 ccttggagcc tacctagact cagccggctc tccacgcttt gcctgaccct gcttgctcaa 3661 ctctacgtct ttgtttcgtt ttctgttctg cgccgttaca gatccaagct gtgaccggcg 3721 cctacgctag acgccaccat ggagagcgac gagagcggcc tgcccgccat ggagatcgag 3781 tgccgcatca ccggcaccct gaacggcgtg gagttcgagc tggtgggcgg cggagagggc 3841 acccccaagc agggccgcat gaccaacaag atgaagagca ccaaaggcgc cctgaccttc 3901 agcccctacc tgctgagcca cgtgatgggc tacggcttct accacttcgg cacctacccc 3961 agcggctacg agaacccctt cctgcacgcc atcaacaacg gcggctacac caacacccgc 4021 atcgagaagt acgaggacgg cggcgtgctg cacgtgagct tcagctaccg ctacgaggcc 4081 ggccgcgtga tcggcgactt caaggtggtg ggcaccggct tccccgagga cagcgtgatc 4141 ttcaccgaca agatcatccg cagcaacgcc accgtggagc acctgcaccc catgggcgat 4201 aacgtgctgg tgggcagctt cgcccgcacc ttcagcctgc gcgacggcgg ctactacagc 4261 ttcgtggtgg acagccacat gcacttcaag agcgccatcc accccagcat cctgcagaac 4321 gggggcccca tgttcgcctt ccgccgcgtg gaggagctgc acagcaacac cgagctgggc 4381 atcgtggagt accagcacgc cttcaagacc cccatcgcct tcgccagatc ccgcgctcag 4441 tcgtccaatt ctgccgtgga cggcaccgcc ggacccggct ccaccggatc tcgcgagggc 4501 agaggaagtc ttctaacatg cggtgacgtg gaggagaatc ccggccctat gaccgagtac 4561 aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg caccctcgcc 4621 gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg ccacatcgag 4681 cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg 4741 tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg 4801 ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc 4861 gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc 4921 ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg 4981 ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg 5041 ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg 5101 cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgaaa tcaacctctg 5161 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta 5221 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt 5281 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc 5341 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt 5401 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg 5461 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac 5521 aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc 5581 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac 5641 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct 5701 cagacgagtc ggatctccct ttgggccgcc tccccgcctg gtacctttaa gaccaatgac 5761 ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct 5821 aattcactcc caacgaagat aagatctgct ttttgcttgt actgggtctc tctggttaga 5881 ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata 5941 aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta 6001 gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtagta gttcatgtca 6061 tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggaactt 6121 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa 6181 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca 6241 tgtctggctc tagctatccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt 6301 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc 6361 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctagacttt 6421 tgcagagacc aaattcgtaa tcatgtcata gctgtttcct gtgtgaaatt gttatccgct 6481 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg 6541 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct 6601 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg 6661 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc 6721 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg 6781 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct 6841 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca 6901 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct 6961 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc 7021 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt 7081 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc 7141 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc 7201 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg 7261 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc 7321 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag 7381 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga 7441 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat 7501 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag 7561 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat 7621 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc 7681 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat 7741 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag 7801 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg 7861 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc 7921 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca 7981 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg 8041 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc 8101 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta 8161 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc 8221 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg 8281 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc 8341 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc 8401 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat 8461 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag 8521 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc 8581 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa 8641 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg 8701 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca 8761 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc 8821 atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt 8881 aaggagaaaa taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg 8941 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag 9001 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag 9061 tgccaagctg //