LOCUS V046678 10034 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V046678 VERSION V046678 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..1920 /note="central polypurine tract and central termination sequence of HIV-1" /label="cPPT/CTS" promoter 2010..2213 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" regulatory 2296..2305 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 3166..3219 /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 3220..4206 /label="CD86(NM_175862)" /note="CD86(NM_175862)" /gene="CD86" CDS 4243..4953 /codon_start=1 /note="mammalian codon-optimized" /product="enhanced monomeric blue fluorescent protein (Subach et al., 2011)" /transl_table=1 /translation="MVSKGEELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTM RIKVVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG VLTATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRND MALKLVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAV ARYCDLPSKLGHKLN" /label="mTagBFP2" promoter 5004..5215 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 5228..5496 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 5523..6122 /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 6129..6717 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6791..7024 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 7096..7217 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 7236..7371 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(7409..7425) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7433..7449 /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(7457..7487) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7502..7523 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7811..8399) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8570..9430) /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(9431..9535) /gene="bla" /label="AmpR promoter" primer_bind 10009..10025 /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 attacaaaat tcaaaatttt 1921 atcgatacta gtattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca 1981 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc 2041 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga 2101 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat 2161 tgacgcaaat gggcggtagg cgtgtacggt gggaggttta tataagcaga gctcgtttag 2221 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagattct 2281 agagctagcg aattcgccac catgggccac acacggaggc agggaacatc accatccaag 2341 tgtccatacc tcaatttctt tcagctcttg gtgctggctg gtctttctca cttctgttca 2401 ggtgttatcc acgtgaccaa ggaagtgaaa gaagtggcaa cgctgtcctg tggtcacaat 2461 gtttctgttg aagagctggc acaaactcgc atctactggc aaaaggagaa gaaaatggtg 2521 ctgactatga tgtctgggga catgaatata tggcccgagt acaagaaccg gaccatcttt 2581 gatatcacta ataacctctc cattgtgatc ctggctctgc gcccatctga cgagggcaca 2641 tacgagtgtg ttgttctgaa gtatgaaaaa gacgctttca agcgggaaca cctggctgaa 2701 gtgacgttat cagtcaaagc tgacttccct acacctagta tatctgactt tgaaattcca 2761 acttctaata ttagaaggat aatttgctca acctctggag gttttccaga gcctcacctc 2821 tcctggttgg aaaatggaga agaattaaat gccatcaaca caacagtttc ccaagatcct 2881 gaaactgagc tctatgctgt tagcagcaaa ctggatttca atatgacaac caaccacagc 2941 ttcatgtgtc tcatcaagta tggacattta agagtgaatc agaccttcaa ctggaataca 3001 accaagcaag agcattttcc tgataacctg ctcccatcct gggccattac cttaatctca 3061 gtaaatggaa tttttgtgat atgctgcctg acctactgct ttgccccaag atgcagagag 3121 agaaggagga atgagagatt gagaagggaa agtgtacgcc ctgtagaggg cagaggaagt 3181 cttctaacat gcggtgacgt ggaggagaat cccggcccta tggatcccca gtgcactatg 3241 ggactgagta acattctctt tgtgatggcc ttcctgctct ctggtgctgc tcctctgaag 3301 attcaagctt atttcaatga gactgcagac ctgccatgcc aatttgcaaa ctctcaaaac 3361 caaagcctga gtgagctagt agtattttgg caggaccagg aaaacttggt tctgaatgag 3421 gtatacttag gcaaagagaa atttgacagt gttcattcca agtatatggg ccgcacaagt 3481 tttgattcgg acagttggac cctgagactt cacaatcttc agatcaagga caagggcttg 3541 tatcaatgta tcatccatca caaaaagccc acaggaatga ttcgcatcca ccagatgaat 3601 tctgaactgt cagtgcttgc taacttcagt caacctgaaa tagtaccaat ttctaatata 3661 acagaaaatg tgtacataaa tttgacctgc tcatctatac acggttaccc agaacctaag 3721 aagatgagtg ttttgctaag aaccaagaat tcaactatcg agtatgatgg tgttatgcag 3781 aaatctcaag ataatgtcac agaactgtac gacgtttcca tcagcttgtc tgtttcattc 3841 cctgatgtta cgagcaatat gaccatcttc tgtattctgg aaactgacaa gacgcggctt 3901 ttatcttcac ctttctctat agagcttgag gaccctcagc ctcccccaga ccacattcct 3961 tggattacag ctgtacttcc aacagttatt atatgtgtga tggttttctg tctaattcta 4021 tggaaatgga agaagaagaa gcggcctcgc aactcttata aatgtggaac caacacaatg 4081 gagagggaag agagtgaaca gaccaagaaa agagaaaaaa tccatatacc tgaaagatct 4141 gatgaagccc agcgtgtttt taaaagttcg aagacatctt catgcgacaa aagtgataca 4201 tgttttctcg agggtggagg cggatcagga ggcggtggat ctatggtgtc taagggcgaa 4261 gagctgatta aggagaacat gcacatgaag ctgtacatgg agggcaccgt ggacaaccat 4321 cacttcaagt gcacatccga gggcgaaggc aagccctacg agggcaccca gaccatgaga 4381 atcaaggtgg tcgagggcgg ccctctcccc ttcgccttcg acatcctggc tactagcttc 4441 ctctacggca gcaagacctt catcaaccac acccagggca tccccgactt cttcaagcag 4501 tccttccctg agggcttcac atgggagaga gtcaccacat acgaagacgg gggcgtgctg 4561 accgctaccc aggacaccag cctccaggac ggctgcctca tctacaacgt caagatcaga 4621 ggggtgaact tcacatccaa cggccctgtg atgcagaaga aaacactcgg ctgggaggcc 4681 ttcaccgaga cgctgtaccc cgctgacggc ggcctggaag gcagaaacga catggccctg 4741 aagctcgtgg gcgggagcca tctgatcgca aacgccaaga ccacatatag atccaagaaa 4801 cccgctaaga acctcaagat gcctggcgtc tactatgtgg actacagact ggaaagaatc 4861 aaggaggcca acaacgagac ctacgtcgag cagcacgagg tggcagtggc cagatactgc 4921 gacctcccta gcaaactggg gcacaagctt aattaaggat ccgcggccgc gaaggatctg 4981 cgatcgctcc ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt 5041 ggggggaggg gtcggcaatt gaacgggtgc ctagagaagg tggcgcgggg taaactggga 5101 aagtgatgtc gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag 5161 tgcagtagtc gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacagctgaa 5221 gcttcgaggg gctcgcatct ctccttcacg cgcccgccgc cctacctgag gccgccatcc 5281 acgccggttg agtcgcgttc tgccgcctcc cgcctgtggt gcctcctgaa ctgcgtccgc 5341 cgtctaggta agtttaaagc tcaggtcgag accgggcctt tgtccggcgc tcccttggag 5401 cctacctaga ctcagccggc tctccacgct ttgcctgacc ctgcttgctc aactctacgt 5461 ctttgtttcg ttttctgttc tgcgccgtta cagatccaag ctgtgaccgg cgcctacgct 5521 agatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc ccccgggccg 5581 tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgacccgg 5641 accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg 5701 acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg 5761 agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg 5821 gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg 5881 agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg 5941 gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc 6001 tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg 6061 ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct 6121 gagtcgacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact 6181 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg 6241 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg 6301 aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa 6361 cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc 6421 ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg 6481 ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt 6541 ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt 6601 cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc 6661 cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg 6721 gtacctttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa 6781 aaggggggac tggaagggct aattcactcc caacgaaaat aagatctgct ttttgcttgt 6841 actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac 6901 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg 6961 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct 7021 agcagtagta gttcatgtca tcttattatt cagtatttat aacttgcaaa gaaatgaata 7081 tcagagagtg agaggaactt gtttattgca gcttataatg gttacaaata aagcaatagc 7141 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa 7201 ctcatcaatg tatcttatca tgtctggctc tagctatccc gcccctaact ccgcccagtt 7261 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg 7321 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctagactttt 7381 gcagagacgg cccaaattcg taatcatggt catagctgtt tcctgtgtga aattgttatc 7441 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct 7501 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa 7561 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta 7621 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc 7681 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg 7741 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt 7801 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa 7861 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct 7921 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc 7981 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg 8041 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct 8101 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag 8161 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga 8221 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga 8281 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg 8341 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag 8401 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag 8461 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat 8521 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct 8581 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac 8641 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa 8701 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg 8761 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt 8821 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca 8881 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt 8941 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct 9001 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg 9061 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg 9121 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg 9181 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa 9241 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt 9301 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt 9361 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt 9421 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca 9481 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat 9541 ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata 9601 aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc 9661 tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca 9721 gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg 9781 cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat 9841 gcgtaaggag aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg 9901 aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg 9961 caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg 10021 ccagtgccaa gctg //