LOCUS V035268 9751 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V035268 VERSION V035268 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..3777 /label="STK4(NM_006282)" /note="STK4(NM_006282)" /gene="STK4" CDS 3784..3849 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3893..4104 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4117..4385 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4414..4423 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4444..5106 /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 5176..5229 /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 5230..5829 /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 5830..6418 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6492..6725 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6797..6918 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6958..7093 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(7126..7142) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7150..7166 /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(7174..7204) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7219..7240 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7528..8116) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8287..9147) /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(9148..9252) /gene="bla" /label="AmpR promoter" primer_bind 9726..9742 /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 agacggtaca gctgaggaac 2341 ccgccgcgcc ggcagctgaa aaagttggat gaagatagtt taaccaaaca accagaagaa 2401 gtatttgatg tcttagagaa acttggagaa gggtcctatg gcagcgtata caaagctatt 2461 cataaagaga ccggccagat tgttgctatt aagcaagttc ctgtggaatc agacctccag 2521 gagataatca aagaaatctc tataatgcag caatgtgaca gccctcatgt agtcaaatat 2581 tatggcagtt attttaagaa cacagactta tggatcgtta tggagtactg tggggctggt 2641 tctgtatctg atatcattcg attacgaaat aaaacgttaa cagaagatga aatagctaca 2701 atattacaat caactcttaa gggacttgaa taccttcatt ttatgagaaa aatacaccga 2761 gatatcaagg caggaaatat tttgctaaat acagaaggac atgcaaaact tgcagatttt 2821 ggggtagcag gtcaacttac agataccatg gccaagcgga atacagtgat aggaacacca 2881 ttttggatgg ctccagaagt gattcaggaa attggataca actgtgtagc agacatctgg 2941 tccctgggaa taactgccat agaaatggct gaaggaaagc ccccttatgc tgatatccat 3001 ccaatgaggg caatcttcat gattcctaca aatcctcctc ccacattccg aaaaccagag 3061 ctatggtcag ataactttac agattttgtg aaacagtgtc ttgtaaagag ccctgagcag 3121 agggccacag ccactcagct cctgcagcac ccatttgtca ggagtgccaa aggagtgtca 3181 atactgcgag acttaattaa tgaagccatg gatgtgaaac tgaaacgcca ggaatcccag 3241 cagcgggaag tggaccagga cgatgaagaa aactcagaag aggatgaaat ggattctggc 3301 acgatggttc gagcagtggg tgatgagatg ggcactgtcc gagtagccag caccatgact 3361 gatggagcca atactatgat tgagcacgat gacacgttgc catcacaact gggcaccatg 3421 gtgatcaatg cagaggatga ggaagaggaa ggaactatga aaagaaggga tgagaccatg 3481 cagcctgcga aaccatcctt tcttgaatat tttgaacaaa aagaaaagga aaaccagatc 3541 aacagctttg gcaagagtgt acctggtcca ctgaaaaatt cttcagattg gaaaatacca 3601 caggatggag actacgagtt tcttaagagt tggacagtgg aggaccttca gaagaggctc 3661 ttggccctgg accccatgat ggagcaggag attgaagaga tccggcagaa gtaccagtcc 3721 aagcggcagc ccatcctgga tgccatagag gctaagaaga gacggcaaca aaacttcgga 3781 tccgactaca aagaccatga cggtgattat aaagatcatg acatcgacta caaggatgac 3841 gatgacaagt agcggccgcg aaggatctgc gatcgctccg gtgcccgtca gtgggcagag 3901 cgcacatcgc ccacagtccc cgagaagttg gggggagggg tcggcaattg aacgggtgcc 3961 tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg tgtactggct ccgccttttt 4021 cccgagggtg ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc 4081 aacgggtttg ccgccagaac acagctgaag cttcgagggg ctcgcatctc tccttcacgc 4141 gcccgccgcc ctacctgagg ccgccatcca cgccggttga gtcgcgttct gccgcctccc 4201 gcctgtggtg cctcctgaac tgcgtccgcc gtctaggtaa gtttaaagct caggtcgaga 4261 ccgggccttt gtccggcgct cccttggagc ctacctagac tcagccggct ctccacgctt 4321 tgcctgaccc tgcttgctca actctacgtc tttgtttcgt tttctgttct gcgccgttac 4381 agatccaagc tgtgaccggc gcctacgcta gacgccacca tggagagcga cgagagcggc 4441 ctgcccgcca tggagatcga gtgccgcatc accggcaccc tgaacggcgt ggagttcgag 4501 ctggtgggcg gcggagaggg cacccccaag cagggccgca tgaccaacaa gatgaagagc 4561 accaaaggcg ccctgacctt cagcccctac ctgctgagcc acgtgatggg ctacggcttc 4621 taccacttcg gcacctaccc cagcggctac gagaacccct tcctgcacgc catcaacaac 4681 ggcggctaca ccaacacccg catcgagaag tacgaggacg gcggcgtgct gcacgtgagc 4741 ttcagctacc gctacgaggc cggccgcgtg atcggcgact tcaaggtggt gggcaccggc 4801 ttccccgagg acagcgtgat cttcaccgac aagatcatcc gcagcaacgc caccgtggag 4861 cacctgcacc ccatgggcga taacgtgctg gtgggcagct tcgcccgcac cttcagcctg 4921 cgcgacggcg gctactacag cttcgtggtg gacagccaca tgcacttcaa gagcgccatc 4981 caccccagca tcctgcagaa cgggggcccc atgttcgcct tccgccgcgt ggaggagctg 5041 cacagcaaca ccgagctggg catcgtggag taccagcacg ccttcaagac ccccatcgcc 5101 ttcgccagat cccgcgctca gtcgtccaat tctgccgtgg acggcaccgc cggacccggc 5161 tccaccggat ctcgcgaggg cagaggaagt cttctaacat gcggtgacgt ggaggagaat 5221 cccggcccta tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc 5281 agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc 5341 gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc 5401 gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc 5461 acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag 5521 ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg 5581 cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag 5641 ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc 5701 gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc 5761 gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc 5821 ggtgcctgaa atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac 5881 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt 5941 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat 6001 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca 6061 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc 6121 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg 6181 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct 6241 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct 6301 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt 6361 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct 6421 ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga 6481 aaagggggga ctggaagggc taattcactc ccaacgaaga taagatctgc tttttgcttg 6541 tactgggtct ctctggttag accagatctg agcctgggag ctctctggct aactagggaa 6601 cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct 6661 gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc 6721 tagcagtagt agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat 6781 atcagagagt gagaggaact tgtttattgc agcttataat ggttacaaat aaagcaatag 6841 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa 6901 actcatcaat gtatcttatc atgtctggct ctagctatcc cgcccctaac tccgcccatc 6961 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt 7021 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc 7081 ttttttggag gcctagactt ttgcagagac caaattcgta atcatgtcat agctgtttcc 7141 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg 7201 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc 7261 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg 7321 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc 7381 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac 7441 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa 7501 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca 7561 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc 7621 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata 7681 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta 7741 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca 7801 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga 7861 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg 7921 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg 7981 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg 8041 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag 8101 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa 8161 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat 8221 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc 8281 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc 8341 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc 8401 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc 8461 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc 8521 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt 8581 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc 8641 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa 8701 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt 8761 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg 8821 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc 8881 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa 8941 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt 9001 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt 9061 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag 9121 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta 9181 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat 9241 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat 9301 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg 9361 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta 9421 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg 9481 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg 9541 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag 9601 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc 9661 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg 9721 acgttgtaaa acgacggcca gtgccaagct g //