LOCUS V039258 9505 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V039258 VERSION V039258 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..3531 /label="CTH(NM_001902)" /note="CTH(NM_001902)" /gene="CTH" CDS 3538..3603 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3647..3858 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3871..4139 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4168..4177 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4198..4860 /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 4930..4983 /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 4984..5583 /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 5584..6172 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6246..6479 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6551..6672 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6712..6847 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6880..6896) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6904..6920 /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(6928..6958) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6973..6994 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7282..7870) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8041..8901) /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(8902..9006) /gene="bla" /label="AmpR promoter" primer_bind 9480..9496 /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 aggaaaaaga cgcctcctca 2341 caaggtttcc tgccacactt ccaacatttc gccacgcagg cgatccatgt gggccaggat 2401 ccagagcaat ggacctccag ggctgtagtg ccccccatct cactgtccac cacgttcaag 2461 caaggggcgc ctggccagca ctcgggtttt gaatatagcc gttctggaaa tcccactagg 2521 aattgccttg aaaaagcagt ggcagcactg gatggggcta agtactgttt ggcctttgct 2581 tcaggtttag cagccactgt aactattacc catcttttaa aagcaggaga ccaaattatt 2641 tgtatggatg atgtgtatgg aggtacaaac aggtacttca ggcaagtggc atctgaattt 2701 ggattaaaga tttcttttgt tgattgttcc aaaatcaaat tactagaggc agcaattaca 2761 ccagaaacca agcttgtttg gatcgaaacc cccacaaacc ccacccagaa ggtgattgac 2821 attgaaggct gtgcacatat tgtccataag catggagaca ttattttggt cgtggataac 2881 acttttatgt caccatattt ccagcgccct ttggctctgg gagctgatat ttctatgtat 2941 tctgcaacaa aatacatgaa tggccacagt gatgttgtaa tgggcctggt gtctgttaat 3001 tgtgaaagcc ttcataatag acttcgtttc ttgcaaaact ctcttggagc agttccatct 3061 cctattgatt gttacctctg caatcgaggt ctgaagactc tacatgtccg aatggaaaag 3121 catttcaaaa acggaatggc agttgcccag ttcctggaat ctaatccttg ggtagaaaag 3181 gttatttatc ctgggctgcc ctctcatcca cagcatgagt tggtgaagcg tcagtgtaca 3241 ggttgtacag ggatggtcac cttttatatt aagggcactc ttcagcatgc tgagattttc 3301 ctcaagaacc taaagctatt tactctggcc gagagcttgg gaggattcga aagccttgct 3361 gagcttccgg caatcatgac tcatgcatca gttcttaaga atgacagaga tgtccttgga 3421 attagtgaca cactgattcg actttctgtg ggcttagagg atgaggaaga cctactggaa 3481 gatctagatc aagctttgaa ggcagcacac cctccaagtg gaagtcacag cggatccgac 3541 tacaaagacc atgacggtga ttataaagat catgacatcg actacaagga tgacgatgac 3601 aagtagcggc cgcgaaggat ctgcgatcgc tccggtgccc gtcagtgggc agagcgcaca 3661 tcgcccacag tccccgagaa gttgggggga ggggtcggca attgaacggg tgcctagaga 3721 aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag 3781 ggtgggggag aaccgtatat aagtgcagta gtcgccgtga acgttctttt tcgcaacggg 3841 tttgccgcca gaacacagct gaagcttcga ggggctcgca tctctccttc acgcgcccgc 3901 cgccctacct gaggccgcca tccacgccgg ttgagtcgcg ttctgccgcc tcccgcctgt 3961 ggtgcctcct gaactgcgtc cgccgtctag gtaagtttaa agctcaggtc gagaccgggc 4021 ctttgtccgg cgctcccttg gagcctacct agactcagcc ggctctccac gctttgcctg 4081 accctgcttg ctcaactcta cgtctttgtt tcgttttctg ttctgcgccg ttacagatcc 4141 aagctgtgac cggcgcctac gctagacgcc accatggaga gcgacgagag cggcctgccc 4201 gccatggaga tcgagtgccg catcaccggc accctgaacg gcgtggagtt cgagctggtg 4261 ggcggcggag agggcacccc caagcagggc cgcatgacca acaagatgaa gagcaccaaa 4321 ggcgccctga ccttcagccc ctacctgctg agccacgtga tgggctacgg cttctaccac 4381 ttcggcacct accccagcgg ctacgagaac cccttcctgc acgccatcaa caacggcggc 4441 tacaccaaca cccgcatcga gaagtacgag gacggcggcg tgctgcacgt gagcttcagc 4501 taccgctacg aggccggccg cgtgatcggc gacttcaagg tggtgggcac cggcttcccc 4561 gaggacagcg tgatcttcac cgacaagatc atccgcagca acgccaccgt ggagcacctg 4621 caccccatgg gcgataacgt gctggtgggc agcttcgccc gcaccttcag cctgcgcgac 4681 ggcggctact acagcttcgt ggtggacagc cacatgcact tcaagagcgc catccacccc 4741 agcatcctgc agaacggggg ccccatgttc gccttccgcc gcgtggagga gctgcacagc 4801 aacaccgagc tgggcatcgt ggagtaccag cacgccttca agacccccat cgccttcgcc 4861 agatcccgcg ctcagtcgtc caattctgcc gtggacggca ccgccggacc cggctccacc 4921 ggatctcgcg agggcagagg aagtcttcta acatgcggtg acgtggagga gaatcccggc 4981 cctatgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc 5041 gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgatccg 5101 gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc 5161 gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg 5221 gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc 5281 ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag 5341 gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg 5401 ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc 5461 ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc 5521 gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc 5581 tgaaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt 5641 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc 5701 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag 5761 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc 5821 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc 5881 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg 5941 ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg 6001 ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc 6061 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt 6121 cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc gcctggtacc 6181 tttaagacca atgacttaca aggcagctgt agatcttagc cactttttaa aagaaaaggg 6241 gggactggaa gggctaattc actcccaacg aagataagat ctgctttttg cttgtactgg 6301 gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact 6361 gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg 6421 tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag 6481 tagtagttca tgtcatctta ttattcagta tttataactt gcaaagaaat gaatatcaga 6541 gagtgagagg aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac 6601 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat 6661 caatgtatct tatcatgtct ggctctagct atcccgcccc taactccgcc catcccgccc 6721 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat 6781 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt 6841 ggaggcctag acttttgcag agaccaaatt cgtaatcatg tcatagctgt ttcctgtgtg 6901 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc 6961 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt 7021 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg 7081 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt 7141 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc 7201 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa 7261 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa 7321 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc 7381 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc 7441 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag 7501 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga 7561 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc 7621 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac 7681 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg 7741 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca 7801 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa 7861 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa 7921 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt 7981 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag 8041 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat 8101 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc 8161 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa 8221 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca 8281 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa 8341 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt 8401 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc 8461 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact 8521 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc 8581 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg 8641 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct 8701 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc 8761 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag 8821 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac 8881 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg 8941 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt 9001 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac 9061 attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga 9121 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga 9181 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg 9241 gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat 9301 accgcacaga tgcgtaagga gaaaataccg catcaggcgc cattcgccat tcaggctgcg 9361 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg 9421 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg 9481 taaaacgacg gccagtgcca agctg //