LOCUS V037729 8864 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V037729 VERSION V037729 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..2949 /label="Rac1(NM_001347530)" /note="Rac1(NM_001347530)" /gene="Rac1" CDS 2956..2979 /codon_start=1 /product="8xHis affinity tag" /transl_table=1 /translation="HHHHHHHH" /label="8xHis" promoter 3023..3234 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3247..3515 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 3544..3553 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 3574..4236 /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 4306..4359 /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 4360..4959 /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 4960..5547 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5621..5854 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 5926..6047 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6066..6201 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6239..6255) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6263..6279 /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(6287..6317) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6332..6353 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6641..7229) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7400..8260) /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(8261..8365) /gene="bla" /label="AmpR promoter" primer_bind 8839..8855 /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 aggccatcaa gtgtgtggtg 2341 gtgggagacg gagctgttgg taaaacctgc ctgctcatca gttacacgac caatgcattt 2401 cctggagagt acatccccac cgtctttgac aactattctg ccaatgttat ggtagatgga 2461 aaaccagtga atctgggcct atgggacaca gctggacaag aagattatga cagattgcgt 2521 cccctctcct acccgcagac agttggagac acatgtggta aagatagacc ctccaggggc 2581 aaagacaagc cgattgccga cgtgttctta atttgctttt cccttgtgag tcctgcatca 2641 tttgaaaatg tccgtgcaaa gtggtatcct gaagtgcgac accactgtcc caatactcct 2701 atcatcctcg tggggacgaa gcttgatctt agggatgata aggacaccat tgagaagctg 2761 aaggagaaga agctgactcc catcacctac ccgcaggggc tggccatggc gaaagagatc 2821 ggtgctgtca aatacctgga gtgctcagct ctcacacagc gaggactcaa gacagtgttt 2881 gacgaagcta tccgagcggt tctctgtccc cctcctgtca agaagaggaa gagaaaatgc 2941 ctgctgttgg gatcccacca tcaccatcac catcaccatt agcggccgcg aaggatctgc 3001 gatcgctccg gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc cgagaagttg 3061 gggggagggg tcggcaattg aacgggtgcc tagagaaggt ggcgcggggt aaactgggaa 3121 agtgatgtcg tgtactggct ccgccttttt cccgagggtg ggggagaacc gtatataagt 3181 gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg ccgccagaac acagctgaag 3241 cttcgagggg ctcgcatctc tccttcacgc gcccgccgcc ctacctgagg ccgccatcca 3301 cgccggttga gtcgcgttct gccgcctccc gcctgtggtg cctcctgaac tgcgtccgcc 3361 gtctaggtaa gtttaaagct caggtcgaga ccgggccttt gtccggcgct cccttggagc 3421 ctacctagac tcagccggct ctccacgctt tgcctgaccc tgcttgctca actctacgtc 3481 tttgtttcgt tttctgttct gcgccgttac agatccaagc tgtgaccggc gcctacgcta 3541 gacgccacca tggagagcga cgagagcggc ctgcccgcca tggagatcga gtgccgcatc 3601 accggcaccc tgaacggcgt ggagttcgag ctggtgggcg gcggagaggg cacccccaag 3661 cagggccgca tgaccaacaa gatgaagagc accaaaggcg ccctgacctt cagcccctac 3721 ctgctgagcc acgtgatggg ctacggcttc taccacttcg gcacctaccc cagcggctac 3781 gagaacccct tcctgcacgc catcaacaac ggcggctaca ccaacacccg catcgagaag 3841 tacgaggacg gcggcgtgct gcacgtgagc ttcagctacc gctacgaggc cggccgcgtg 3901 atcggcgact tcaaggtggt gggcaccggc ttccccgagg acagcgtgat cttcaccgac 3961 aagatcatcc gcagcaacgc caccgtggag cacctgcacc ccatgggcga taacgtgctg 4021 gtgggcagct tcgcccgcac cttcagcctg cgcgacggcg gctactacag cttcgtggtg 4081 gacagccaca tgcacttcaa gagcgccatc caccccagca tcctgcagaa cgggggcccc 4141 atgttcgcct tccgccgcgt ggaggagctg cacagcaaca ccgagctggg catcgtggag 4201 taccagcacg ccttcaagac ccccatcgcc ttcgccagat cccgcgctca gtcgtccaat 4261 tctgccgtgg acggcaccgc cggacccggc tccaccggat ctcgcgaggg cagaggaagt 4321 cttctaacat gcggtgacgt ggaggagaat cccggcccta tgaccgagta caagcccacg 4381 gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc 4441 gccgactacc ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc 4501 gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg 4561 gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg 4621 ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa 4681 cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc 4741 gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga 4801 gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac 4861 ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga 4921 ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgaa atcaacctct ggattacaaa 4981 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac 5041 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc 5101 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt 5161 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc 5221 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc 5281 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg 5341 gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt 5401 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc 5461 cgcggcctgc tgccggctct gcggcctctt ccgcgtctcc gccttcgccc tcagacgagt 5521 cggatctccc tttggccgcc tccccgcctg gtacctttaa gaccaatgac ttacaaggca 5581 gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc 5641 caacgaaaat aagatctgct ttttgcttgt actgggtctc tctggttaga ccagatctga 5701 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct 5761 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc 5821 agaccctttt agtcagtgtg gaaaatctct agcagtagta gttcatgtca tcttattatt 5881 cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggaactt gtttattgca 5941 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt 6001 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggctc 6061 tagctatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa 6121 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt 6181 gaggaggctt ttttggaggc ctagactttt gcagagacgg cccaaattcg taatcatggt 6241 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg 6301 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt 6361 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg 6421 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg 6481 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa 6541 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc 6601 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc 6661 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat 6721 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc 6781 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct 6841 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg 6901 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc 6961 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga 7021 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa 7081 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta 7141 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc 7201 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg 7261 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga 7321 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg 7381 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct 7441 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg 7501 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc 7561 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa 7621 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc 7681 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt 7741 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc 7801 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt 7861 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc 7921 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt 7981 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata 8041 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga 8101 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag 8161 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa 8221 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt 8281 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga 8341 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag 8401 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc 8461 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca 8521 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg 8581 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc 8641 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc 8701 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat 8761 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt 8821 tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa gctg //