LOCUS V028856 9424 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V028856 VERSION V028856 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..3450 /label="Sting1(NM_028261)" /note="Sting1(NM_028261)" /gene="Sting1" CDS 3457..3522 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3566..3777 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3790..4058 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4087..4096 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4117..4779 /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 4849..4902 /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 4903..5502 /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 5503..6091 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6165..6398 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6470..6591 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6631..6766 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6799..6815) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6823..6839 /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(6847..6877) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6892..6913 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7201..7789) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7960..8820) /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(8821..8925) /gene="bla" /label="AmpR promoter" primer_bind 9399..9415 /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 catactccaa cctgcatcca 2341 gccatcccac ggcccagagg tcaccgctcc aaatatgtag ccctcatctt tctggtggcc 2401 agcctgatga tcctttgggt ggcaaaggat ccaccaaatc acactctgaa gtacctagca 2461 cttcacctag cctcgcacga acttggacta ctgttgaaaa acctctgctg tctggctgaa 2521 gagctgtgcc atgtccagtc caggtaccag ggcagctact ggaaggctgt gcgcgcctgc 2581 ctgggatgcc ccatccactg tatggctatg attctactat cgtcttattt ctatttcctc 2641 caaaacactg ctgacatata cctcagttgg atgtttggcc ttctggtcct ctataagtcc 2701 ctaagcatgc tcctgggcct tcagagcttg actccagcgg aagtctctgc agtctgtgaa 2761 gaaaagaagt taaatgttgc ccacgggctg gcctggtcat actacattgg gtacttgcgg 2821 ttgatcttac cagggctcca ggcccggatc cgaatgttca atcagctaca taacaacatg 2881 ctcagtggtg cagggagccg aagactgtac atcctctttc cattggactg tggggtgcct 2941 gacaacctga gtgtagttga ccccaacatt cgattccgag atatgctgcc ccagcaaaac 3001 atcgaccgtg ctggcatcaa gaatcgggtt tattccaaca gcgtctacga gattctggag 3061 aacggacagc cagcaggcgt ctgtatcctg gagtacgcca cccccttgca gaccctgttt 3121 gccatgtcac aggatgccaa agctggcttc agtcgggagg atcggcttga gcaggctaaa 3181 ctcttctgcc ggacacttga ggaaatcctg gaagatgtcc ccgagtctcg aaataactgc 3241 cgcctcattg tctaccaaga acccacagac ggaaacagtt tctcactgtc tcaggaggtg 3301 ctccggcaca ttcgtcagga agaaaaggag gaggttacca tgaatgcccc catgacctca 3361 gtggcacctc ctccctccgt actgtcccaa gagccaagac tcctcatcag tggtatggat 3421 cagcctctcc cactccgcac tgacctcatc ggatccgact acaaagacca tgacggtgat 3481 tataaagatc atgacatcga ctacaaggat gacgatgaca agtagcggcc gcgaaggatc 3541 tgcgatcgct ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag 3601 ttggggggag gggtcggcaa ttgaacgggt gcctagagaa ggtggcgcgg ggtaaactgg 3661 gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata 3721 agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacagctg 3781 aagcttcgag gggctcgcat ctctccttca cgcgcccgcc gccctacctg aggccgccat 3841 ccacgccggt tgagtcgcgt tctgccgcct cccgcctgtg gtgcctcctg aactgcgtcc 3901 gccgtctagg taagtttaaa gctcaggtcg agaccgggcc tttgtccggc gctcccttgg 3961 agcctaccta gactcagccg gctctccacg ctttgcctga ccctgcttgc tcaactctac 4021 gtctttgttt cgttttctgt tctgcgccgt tacagatcca agctgtgacc ggcgcctacg 4081 ctagacgcca ccatggagag cgacgagagc ggcctgcccg ccatggagat cgagtgccgc 4141 atcaccggca ccctgaacgg cgtggagttc gagctggtgg gcggcggaga gggcaccccc 4201 aagcagggcc gcatgaccaa caagatgaag agcaccaaag gcgccctgac cttcagcccc 4261 tacctgctga gccacgtgat gggctacggc ttctaccact tcggcaccta ccccagcggc 4321 tacgagaacc ccttcctgca cgccatcaac aacggcggct acaccaacac ccgcatcgag 4381 aagtacgagg acggcggcgt gctgcacgtg agcttcagct accgctacga ggccggccgc 4441 gtgatcggcg acttcaaggt ggtgggcacc ggcttccccg aggacagcgt gatcttcacc 4501 gacaagatca tccgcagcaa cgccaccgtg gagcacctgc accccatggg cgataacgtg 4561 ctggtgggca gcttcgcccg caccttcagc ctgcgcgacg gcggctacta cagcttcgtg 4621 gtggacagcc acatgcactt caagagcgcc atccacccca gcatcctgca gaacgggggc 4681 cccatgttcg ccttccgccg cgtggaggag ctgcacagca acaccgagct gggcatcgtg 4741 gagtaccagc acgccttcaa gacccccatc gccttcgcca gatcccgcgc tcagtcgtcc 4801 aattctgccg tggacggcac cgccggaccc ggctccaccg gatctcgcga gggcagagga 4861 agtcttctaa catgcggtga cgtggaggag aatcccggcc ctatgaccga gtacaagccc 4921 acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg 4981 ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc 5041 accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc 5101 gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg 5161 gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag 5221 caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc 5281 accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc 5341 ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc 5401 aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa 5461 ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gaaatcaacc tctggattac 5521 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga 5581 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc 5641 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa 5701 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc 5761 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc 5821 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc 5881 gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg 5941 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct 6001 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg 6061 agtcggatct ccctttgggc cgcctccccg cctggtacct ttaagaccaa tgacttacaa 6121 ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca 6181 ctcccaacga agataagatc tgctttttgc ttgtactggg tctctctggt tagaccagat 6241 ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt 6301 gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc 6361 cctcagaccc ttttagtcag tgtggaaaat ctctagcagt agtagttcat gtcatcttat 6421 tattcagtat ttataacttg caaagaaatg aatatcagag agtgagagga acttgtttat 6481 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt 6541 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg 6601 gctctagcta tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc 6661 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg 6721 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctaga cttttgcaga 6781 gaccaaattc gtaatcatgt catagctgtt tcctgtgtga aattgttatc cgctcacaat 6841 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag 6901 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg 6961 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc 7021 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc 7081 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa 7141 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt 7201 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg 7261 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg 7321 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag 7381 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc 7441 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa 7501 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg 7561 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc 7621 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac 7681 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg 7741 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt 7801 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt 7861 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa 7921 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga 7981 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt 8041 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg 8101 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga 8161 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga 8221 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg 8281 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc 8341 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc 8401 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca 8461 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac 8521 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg 8581 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc 8641 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg 8701 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac 8761 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat 8821 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata 8881 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa 8941 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg 9001 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat 9061 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg 9121 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga 9181 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag 9241 aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc 9301 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt 9361 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa 9421 gctg //