LOCUS V038317 9304 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V038317 VERSION V038317 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..3330 /label="Mrgprb2(NM_175531)" /note="Mrgprb2(NM_175531)" /gene="Mrgprb2" CDS 3337..3402 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3446..3657 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3670..3938 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 3967..3976 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 3997..4659 /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 4729..4782 /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 4783..5382 /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 5383..5971 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6045..6278 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6350..6471 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6511..6646 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6679..6695) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6703..6719 /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(6727..6757) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6772..6793 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7081..7669) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7840..8700) /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(8701..8805) /gene="bla" /label="AmpR promoter" primer_bind 9279..9295 /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 gtggagattt cctaatcaag 2341 aatctaagca cctcagcctg gaaaacgaac atcacagtgc tgaatggaag ctactacatc 2401 gatacttcag tttgtgtcac caggaaccaa gccatgattt tgctttccat catcatttcc 2461 ctggttggga tgggactaaa tgccatagtg ctgtggttcc tgggcatccg tatgcacacg 2521 aatgccttca ctgtctacat tctcaacctg gctatggctg actttcttta cctgtgctct 2581 cagtttgtaa tttgtcttct tattgccttt tatatcttct actcaattga catcaacatc 2641 cctttggttc tttatgttgt gccaatattt gcttatcttt caggtctgag cattctcagc 2701 accattagca ttgagcgctg cttgtctgta atatggccca tttggtatcg ctgtaaacgt 2761 ccaagacaca catcagctat cacatgtttt gtgctttggg ttatgtcctt attgttgggt 2821 ctcctggaag ggaaggcatg tggcttactg tttaatagct ttgactctta ttggtgtgaa 2881 acatttgatg ttatcactaa tatatggtca gttgtttttt ttggtgttct ctgtgggtct 2941 agcctcaccc tgcttgtcag gatcttctgt ggctcacagc gaattcctat gaccaggctg 3001 tatgtgacta ttacactcac agtcttggtc ttcctgatct ttggtcttcc ctttgggatc 3061 tattggatac tctatcagtg gattagcaat ttttattatg ttgaaatttg taatttttat 3121 cttgagatac tattcctatc ctgtgttaac agctgtatga accccatcat ttatttcctt 3181 gttggctcca ttaggcaccg aaggttcagg cggaagactc tcaagctact tctgcagaga 3241 gccatgcaag acacccctga ggaggaacaa agtggaaata agagttcttc agaacaccct 3301 gaagaactgg aaactgttca gagctgcagc ggatccgact acaaagacca tgacggtgat 3361 tataaagatc atgacatcga ctacaaggat gacgatgaca agtagcggcc gcgaaggatc 3421 tgcgatcgct ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag 3481 ttggggggag gggtcggcaa ttgaacgggt gcctagagaa ggtggcgcgg ggtaaactgg 3541 gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata 3601 agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacagctg 3661 aagcttcgag gggctcgcat ctctccttca cgcgcccgcc gccctacctg aggccgccat 3721 ccacgccggt tgagtcgcgt tctgccgcct cccgcctgtg gtgcctcctg aactgcgtcc 3781 gccgtctagg taagtttaaa gctcaggtcg agaccgggcc tttgtccggc gctcccttgg 3841 agcctaccta gactcagccg gctctccacg ctttgcctga ccctgcttgc tcaactctac 3901 gtctttgttt cgttttctgt tctgcgccgt tacagatcca agctgtgacc ggcgcctacg 3961 ctagacgcca ccatggagag cgacgagagc ggcctgcccg ccatggagat cgagtgccgc 4021 atcaccggca ccctgaacgg cgtggagttc gagctggtgg gcggcggaga gggcaccccc 4081 aagcagggcc gcatgaccaa caagatgaag agcaccaaag gcgccctgac cttcagcccc 4141 tacctgctga gccacgtgat gggctacggc ttctaccact tcggcaccta ccccagcggc 4201 tacgagaacc ccttcctgca cgccatcaac aacggcggct acaccaacac ccgcatcgag 4261 aagtacgagg acggcggcgt gctgcacgtg agcttcagct accgctacga ggccggccgc 4321 gtgatcggcg acttcaaggt ggtgggcacc ggcttccccg aggacagcgt gatcttcacc 4381 gacaagatca tccgcagcaa cgccaccgtg gagcacctgc accccatggg cgataacgtg 4441 ctggtgggca gcttcgcccg caccttcagc ctgcgcgacg gcggctacta cagcttcgtg 4501 gtggacagcc acatgcactt caagagcgcc atccacccca gcatcctgca gaacgggggc 4561 cccatgttcg ccttccgccg cgtggaggag ctgcacagca acaccgagct gggcatcgtg 4621 gagtaccagc acgccttcaa gacccccatc gccttcgcca gatcccgcgc tcagtcgtcc 4681 aattctgccg tggacggcac cgccggaccc ggctccaccg gatctcgcga gggcagagga 4741 agtcttctaa catgcggtga cgtggaggag aatcccggcc ctatgaccga gtacaagccc 4801 acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg 4861 ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc 4921 accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc 4981 gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg 5041 gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag 5101 caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc 5161 accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc 5221 ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc 5281 aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa 5341 ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gaaatcaacc tctggattac 5401 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga 5461 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc 5521 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa 5581 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc 5641 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc 5701 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc 5761 gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg 5821 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct 5881 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg 5941 agtcggatct ccctttgggc cgcctccccg cctggtacct ttaagaccaa tgacttacaa 6001 ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca 6061 ctcccaacga agataagatc tgctttttgc ttgtactggg tctctctggt tagaccagat 6121 ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt 6181 gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc 6241 cctcagaccc ttttagtcag tgtggaaaat ctctagcagt agtagttcat gtcatcttat 6301 tattcagtat ttataacttg caaagaaatg aatatcagag agtgagagga acttgtttat 6361 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt 6421 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg 6481 gctctagcta tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc 6541 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg 6601 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctaga cttttgcaga 6661 gaccaaattc gtaatcatgt catagctgtt tcctgtgtga aattgttatc cgctcacaat 6721 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag 6781 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg 6841 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc 6901 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc 6961 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa 7021 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt 7081 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg 7141 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg 7201 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag 7261 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc 7321 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa 7381 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg 7441 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc 7501 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac 7561 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg 7621 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt 7681 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt 7741 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa 7801 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga 7861 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt 7921 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg 7981 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga 8041 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga 8101 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg 8161 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc 8221 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc 8281 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca 8341 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac 8401 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg 8461 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc 8521 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg 8581 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac 8641 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat 8701 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata 8761 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa 8821 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg 8881 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat 8941 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg 9001 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga 9061 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag 9121 aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc 9181 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt 9241 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa 9301 gctg //