LOCUS V026537 9724 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V026537 VERSION V026537 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..3750 /label="IFIT1(NM_001548)" /note="IFIT1(NM_001548)" /gene="IFIT1" CDS 3757..3822 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3866..4077 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4090..4358 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4387..4396 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4417..5079 /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 5149..5202 /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 5203..5802 /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 5803..6391 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6465..6698 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6770..6891 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6931..7066 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(7099..7115) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7123..7139 /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(7147..7177) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7192..7213 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7501..8089) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8260..9120) /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(9121..9225) /gene="bla" /label="AmpR promoter" primer_bind 9699..9715 /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 gtacaaatgg tgatgatcat 2341 caggtcaagg atagtctgga gcaattgaga tgtcacttta catgggagtt atccattgat 2401 gacgatgaaa tgcctgattt agaaaacaga gtcttggatc agattgaatt cctagacacc 2461 aaatacagtg tgggaataca caacctacta gcctatgtga aacacctgaa aggccagaat 2521 gaggaagccc tgaagagctt aaaagaagct gaaaacttaa tgcaggaaga acatgacaac 2581 caagcaaatg tgaggagtct ggtgacctgg ggcaactttg cctggatgta ttaccacatg 2641 ggcagactgg cagaagccca gacttacctg gacaaggtgg agaacatttg caagaagctt 2701 tcaaatccct tccgctatag aatggagtgt ccagaaatag actgtgagga aggatgggcc 2761 ttgctgaagt gtggaggaaa aaattatgaa cgggccaagg cctgctttga aaaggtgctt 2821 gaagtggacc ctgaaaaccc tgaatccagc gctgggtatg cgatctctgc ctatcgcctg 2881 gatggcttta aattagccac aaaaaatcac aagccatttt ctttgcttcc cctaaggcag 2941 gctgtccgct taaatccaga caatggatat attaaggttc tccttgccct gaagcttcag 3001 gatgaaggac aggaagctga aggagaaaag tacattgaag aagctctagc caacatgtcc 3061 tcacagacct atgtctttcg atatgcagcc aagttttacc gaagaaaagg ctctgtggat 3121 aaagctcttg agttattaaa aaaggccttg caggaaacac ccacttctgt cttactgcat 3181 caccagatag ggctttgcta caaggcacaa atgatccaaa tcaaggaggc tacaaaaggg 3241 cagcctagag ggcagaacag agaaaagcta gacaaaatga taagatcagc catatttcat 3301 tttgaatctg cagtggaaaa aaagcccaca tttgaggtgg ctcatctaga cctggcaaga 3361 atgtatatag aagcaggcaa tcacagaaaa gctgaagaga attttcaaaa attgttatgc 3421 atgaaaccag tggtagaaga aacaatgcaa gacatacatt tccactatgg tcggtttcag 3481 gaatttcaaa agaaatctga cgtcaatgca attatccatt atttaaaagc tataaaaata 3541 gaacaggcat cattaacaag ggataaaagt atcaattctt tgaagaaatt ggttttaagg 3601 aaacttcgga gaaaggcatt agatctggaa agcttgagcc tccttgggtt cgtctacaaa 3661 ttggaaggaa atatgaatga agccctggag tactatgagc gggccctgag actggctgct 3721 gactttgaga actctgtgag acaaggtcct ggatccgact acaaagacca tgacggtgat 3781 tataaagatc atgacatcga ctacaaggat gacgatgaca agtagcggcc gcgaaggatc 3841 tgcgatcgct ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag 3901 ttggggggag gggtcggcaa ttgaacgggt gcctagagaa ggtggcgcgg ggtaaactgg 3961 gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata 4021 agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacagctg 4081 aagcttcgag gggctcgcat ctctccttca cgcgcccgcc gccctacctg aggccgccat 4141 ccacgccggt tgagtcgcgt tctgccgcct cccgcctgtg gtgcctcctg aactgcgtcc 4201 gccgtctagg taagtttaaa gctcaggtcg agaccgggcc tttgtccggc gctcccttgg 4261 agcctaccta gactcagccg gctctccacg ctttgcctga ccctgcttgc tcaactctac 4321 gtctttgttt cgttttctgt tctgcgccgt tacagatcca agctgtgacc ggcgcctacg 4381 ctagacgcca ccatggagag cgacgagagc ggcctgcccg ccatggagat cgagtgccgc 4441 atcaccggca ccctgaacgg cgtggagttc gagctggtgg gcggcggaga gggcaccccc 4501 aagcagggcc gcatgaccaa caagatgaag agcaccaaag gcgccctgac cttcagcccc 4561 tacctgctga gccacgtgat gggctacggc ttctaccact tcggcaccta ccccagcggc 4621 tacgagaacc ccttcctgca cgccatcaac aacggcggct acaccaacac ccgcatcgag 4681 aagtacgagg acggcggcgt gctgcacgtg agcttcagct accgctacga ggccggccgc 4741 gtgatcggcg acttcaaggt ggtgggcacc ggcttccccg aggacagcgt gatcttcacc 4801 gacaagatca tccgcagcaa cgccaccgtg gagcacctgc accccatggg cgataacgtg 4861 ctggtgggca gcttcgcccg caccttcagc ctgcgcgacg gcggctacta cagcttcgtg 4921 gtggacagcc acatgcactt caagagcgcc atccacccca gcatcctgca gaacgggggc 4981 cccatgttcg ccttccgccg cgtggaggag ctgcacagca acaccgagct gggcatcgtg 5041 gagtaccagc acgccttcaa gacccccatc gccttcgcca gatcccgcgc tcagtcgtcc 5101 aattctgccg tggacggcac cgccggaccc ggctccaccg gatctcgcga gggcagagga 5161 agtcttctaa catgcggtga cgtggaggag aatcccggcc ctatgaccga gtacaagccc 5221 acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg 5281 ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc 5341 accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc 5401 gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg 5461 gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag 5521 caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc 5581 accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc 5641 ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc 5701 aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa 5761 ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gaaatcaacc tctggattac 5821 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga 5881 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc 5941 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa 6001 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc 6061 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc 6121 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc 6181 gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg 6241 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct 6301 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg 6361 agtcggatct ccctttgggc cgcctccccg cctggtacct ttaagaccaa tgacttacaa 6421 ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca 6481 ctcccaacga agataagatc tgctttttgc ttgtactggg tctctctggt tagaccagat 6541 ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt 6601 gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc 6661 cctcagaccc ttttagtcag tgtggaaaat ctctagcagt agtagttcat gtcatcttat 6721 tattcagtat ttataacttg caaagaaatg aatatcagag agtgagagga acttgtttat 6781 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt 6841 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg 6901 gctctagcta tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc 6961 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg 7021 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctaga cttttgcaga 7081 gaccaaattc gtaatcatgt catagctgtt tcctgtgtga aattgttatc cgctcacaat 7141 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag 7201 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg 7261 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc 7321 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc 7381 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa 7441 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt 7501 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg 7561 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg 7621 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag 7681 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc 7741 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa 7801 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg 7861 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc 7921 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac 7981 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg 8041 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt 8101 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt 8161 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa 8221 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga 8281 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt 8341 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg 8401 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga 8461 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga 8521 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg 8581 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc 8641 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc 8701 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca 8761 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac 8821 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg 8881 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc 8941 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg 9001 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac 9061 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat 9121 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata 9181 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa 9241 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg 9301 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat 9361 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg 9421 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga 9481 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag 9541 aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc 9601 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt 9661 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa 9721 gctg //