LOCUS V028041 8971 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V028041 VERSION V028041 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..2997 /label="Trem2(NM_031254)" /note="Trem2(NM_031254)" /gene="Trem2" CDS 3004..3069 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3113..3324 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3337..3605 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 3634..3643 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 3664..4326 /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 4396..4449 /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 4450..5049 /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 5050..5638 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5712..5945 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6017..6138 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6178..6313 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6346..6362) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6370..6386 /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(6394..6424) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6439..6460 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6748..7336) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7507..8367) /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(8368..8472) /gene="bla" /label="AmpR promoter" primer_bind 8946..8962 /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 gccaccatgg gacctctcca ccagtttctc 2341 ctgctgctga tcacagccct gtcccaagcc ctcaacacca cggtgctgca gggcatggcc 2401 ggccagtcct tgagggtgtc atgtacttat gacgccttga agcactgggg gagacgcaag 2461 gcctggtgtc ggcagctggg tgaggagggc ccatgccagc gtgtggtgag cacacacggt 2521 gtgtggctgc tggccttcct gaagaagcgg aatgggagca cagtcatcgc agatgacacc 2581 cttgctggaa ccgtcaccat cactctgaag aacctccaag ccggtgacgc gggcctctac 2641 cagtgtcaga gtctccgagg ccgagaggct gaggtcctgc agaaagtact ggtggaggtg 2701 ctggaggacc ctctagatga ccaagatgct ggagatctct gggtccccga ggagtcatcg 2761 agtttcgagg gtgcccaagt ggaacacagc acctccagga atcaagagac ctccttccca 2821 cccacctcca ttcttctcct cctggcctgc gttctcctga gcaagtttct tgcagccagc 2881 atcctctggg ctgtggccag gggcaggcag aagccgggaa cacctgtggt cagagggctg 2941 gactgtggcc aagatgctgg gcaccaactt cagatcctca ctggacccgg aggtacggga 3001 tccgactaca aagaccatga cggtgattat aaagatcatg acatcgacta caaggatgac 3061 gatgacaagt agcggccgcg aaggatctgc gatcgctccg gtgcccgtca gtgggcagag 3121 cgcacatcgc ccacagtccc cgagaagttg gggggagggg tcggcaattg aacgggtgcc 3181 tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg tgtactggct ccgccttttt 3241 cccgagggtg ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc 3301 aacgggtttg ccgccagaac acagctgaag cttcgagggg ctcgcatctc tccttcacgc 3361 gcccgccgcc ctacctgagg ccgccatcca cgccggttga gtcgcgttct gccgcctccc 3421 gcctgtggtg cctcctgaac tgcgtccgcc gtctaggtaa gtttaaagct caggtcgaga 3481 ccgggccttt gtccggcgct cccttggagc ctacctagac tcagccggct ctccacgctt 3541 tgcctgaccc tgcttgctca actctacgtc tttgtttcgt tttctgttct gcgccgttac 3601 agatccaagc tgtgaccggc gcctacgcta gacgccacca tggagagcga cgagagcggc 3661 ctgcccgcca tggagatcga gtgccgcatc accggcaccc tgaacggcgt ggagttcgag 3721 ctggtgggcg gcggagaggg cacccccaag cagggccgca tgaccaacaa gatgaagagc 3781 accaaaggcg ccctgacctt cagcccctac ctgctgagcc acgtgatggg ctacggcttc 3841 taccacttcg gcacctaccc cagcggctac gagaacccct tcctgcacgc catcaacaac 3901 ggcggctaca ccaacacccg catcgagaag tacgaggacg gcggcgtgct gcacgtgagc 3961 ttcagctacc gctacgaggc cggccgcgtg atcggcgact tcaaggtggt gggcaccggc 4021 ttccccgagg acagcgtgat cttcaccgac aagatcatcc gcagcaacgc caccgtggag 4081 cacctgcacc ccatgggcga taacgtgctg gtgggcagct tcgcccgcac cttcagcctg 4141 cgcgacggcg gctactacag cttcgtggtg gacagccaca tgcacttcaa gagcgccatc 4201 caccccagca tcctgcagaa cgggggcccc atgttcgcct tccgccgcgt ggaggagctg 4261 cacagcaaca ccgagctggg catcgtggag taccagcacg ccttcaagac ccccatcgcc 4321 ttcgccagat cccgcgctca gtcgtccaat tctgccgtgg acggcaccgc cggacccggc 4381 tccaccggat ctcgcgaggg cagaggaagt cttctaacat gcggtgacgt ggaggagaat 4441 cccggcccta tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc 4501 agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc 4561 gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc 4621 gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc 4681 acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag 4741 ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg 4801 cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag 4861 ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc 4921 gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc 4981 gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc 5041 ggtgcctgaa atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac 5101 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt 5161 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat 5221 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca 5281 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc 5341 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg 5401 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct 5461 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct 5521 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt 5581 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct 5641 ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga 5701 aaagggggga ctggaagggc taattcactc ccaacgaaga taagatctgc tttttgcttg 5761 tactgggtct ctctggttag accagatctg agcctgggag ctctctggct aactagggaa 5821 cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct 5881 gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc 5941 tagcagtagt agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat 6001 atcagagagt gagaggaact tgtttattgc agcttataat ggttacaaat aaagcaatag 6061 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa 6121 actcatcaat gtatcttatc atgtctggct ctagctatcc cgcccctaac tccgcccatc 6181 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt 6241 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc 6301 ttttttggag gcctagactt ttgcagagac caaattcgta atcatgtcat agctgtttcc 6361 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg 6421 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc 6481 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg 6541 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc 6601 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac 6661 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa 6721 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca 6781 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc 6841 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata 6901 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta 6961 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca 7021 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga 7081 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg 7141 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg 7201 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg 7261 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag 7321 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa 7381 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat 7441 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc 7501 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc 7561 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc 7621 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc 7681 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc 7741 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt 7801 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc 7861 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa 7921 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt 7981 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg 8041 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc 8101 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa 8161 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt 8221 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt 8281 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag 8341 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta 8401 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat 8461 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat 8521 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg 8581 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta 8641 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg 8701 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg 8761 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag 8821 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc 8881 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg 8941 acgttgtaaa acgacggcca gtgccaagct g //