LOCUS V046665 9484 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V046665 VERSION V046665 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..3510 /label="ABHD12(NM_001042472)" /note="ABHD12(NM_001042472)" /gene="ABHD12" CDS 3517..3582 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3626..3837 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3850..4118 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4147..4156 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4177..4839 /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 4909..4962 /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 4963..5562 /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 5563..6151 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6225..6458 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6530..6651 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6691..6826 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6859..6875) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6883..6899 /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(6907..6937) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6952..6973 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7261..7849) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8020..8880) /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(8881..8985) /gene="bla" /label="AmpR promoter" primer_bind 9459..9475 /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 ggaagcggac cgagcccgtc 2341 gccttggagc atgagcgctg cgccgccgcg ggctcgtcct cctccggctc ggccgccgcg 2401 gcgctggacg ccgactgccg cctgaagcag aacctacgcc tgacgggccc ggcggcggct 2461 gagccgcgct gcgcagccga cgcgggaatg aagcgggcgc tgggcaggcg aaagggcgtg 2521 tggttgcgcc tgaggaagat acttttctgt gttttggggt tgtacattgc cattccattt 2581 ctcatcaaac tatgtcctgg aatacaggcc aaactgattt tcttgaattt cgtaagagtt 2641 ccctatttca ttgatttgaa aaaaccacag gatcaaggtt tgaatcacac gtgtaactac 2701 tacctgcagc cagaggaaga cgtgaccatt ggagtctggc acaccgtccc tgcagtctgg 2761 tggaagaacg cccaaggcaa agaccagatg tggtatgagg atgccttggc ttccagccac 2821 cctatcattc tgtacctgca tgggaacgca ggtaccagag gaggcgacca ccgcgtggag 2881 ctttacaagg tgctgagttc ccttggttac catgtggtca cctttgacta cagaggttgg 2941 ggtgactcag tgggaacgcc atctgagcgg ggcatgacct atgacgcact ccacgttttt 3001 gactggatca aagcaagaag tggtgacaac cccgtgtaca tctggggcca ctctctgggc 3061 actggcgtgg cgacaaatct ggtgcggcgc ctctgtgagc gagagacgcc tccagatgcc 3121 cttatattgg aatctccatt cactaatatc cgcgaagaag ctaagagcca tccattttca 3181 gtgatatatc gatacttccc tgggtttgac tggttcttcc ttgatcctat tacaagtagt 3241 ggaattaaat ttgcaaatga tgaaaacgtg aagcacatct cctgtcccct gctcatcctg 3301 cacgctgagg acgacccggt ggtgcccttc cagcttggca gaaagctcta tagcatcgcc 3361 gcaccagctc gaagcttccg agatttcaaa gttcagtttg tgccctttca ttcagacctt 3421 ggctacaggc acaaatacat ttacaagagc cctgagctgc cacggatact gagggaattc 3481 ctggggaagt cggagcctga gcaccagcac ggatccgact acaaagacca tgacggtgat 3541 tataaagatc atgacatcga ctacaaggat gacgatgaca agtagcggcc gcgaaggatc 3601 tgcgatcgct ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag 3661 ttggggggag gggtcggcaa ttgaacgggt gcctagagaa ggtggcgcgg ggtaaactgg 3721 gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata 3781 agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacagctg 3841 aagcttcgag gggctcgcat ctctccttca cgcgcccgcc gccctacctg aggccgccat 3901 ccacgccggt tgagtcgcgt tctgccgcct cccgcctgtg gtgcctcctg aactgcgtcc 3961 gccgtctagg taagtttaaa gctcaggtcg agaccgggcc tttgtccggc gctcccttgg 4021 agcctaccta gactcagccg gctctccacg ctttgcctga ccctgcttgc tcaactctac 4081 gtctttgttt cgttttctgt tctgcgccgt tacagatcca agctgtgacc ggcgcctacg 4141 ctagacgcca ccatggagag cgacgagagc ggcctgcccg ccatggagat cgagtgccgc 4201 atcaccggca ccctgaacgg cgtggagttc gagctggtgg gcggcggaga gggcaccccc 4261 aagcagggcc gcatgaccaa caagatgaag agcaccaaag gcgccctgac cttcagcccc 4321 tacctgctga gccacgtgat gggctacggc ttctaccact tcggcaccta ccccagcggc 4381 tacgagaacc ccttcctgca cgccatcaac aacggcggct acaccaacac ccgcatcgag 4441 aagtacgagg acggcggcgt gctgcacgtg agcttcagct accgctacga ggccggccgc 4501 gtgatcggcg acttcaaggt ggtgggcacc ggcttccccg aggacagcgt gatcttcacc 4561 gacaagatca tccgcagcaa cgccaccgtg gagcacctgc accccatggg cgataacgtg 4621 ctggtgggca gcttcgcccg caccttcagc ctgcgcgacg gcggctacta cagcttcgtg 4681 gtggacagcc acatgcactt caagagcgcc atccacccca gcatcctgca gaacgggggc 4741 cccatgttcg ccttccgccg cgtggaggag ctgcacagca acaccgagct gggcatcgtg 4801 gagtaccagc acgccttcaa gacccccatc gccttcgcca gatcccgcgc tcagtcgtcc 4861 aattctgccg tggacggcac cgccggaccc ggctccaccg gatctcgcga gggcagagga 4921 agtcttctaa catgcggtga cgtggaggag aatcccggcc ctatgaccga gtacaagccc 4981 acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg 5041 ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc 5101 accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc 5161 gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg 5221 gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag 5281 caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc 5341 accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc 5401 ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc 5461 aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa 5521 ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gaaatcaacc tctggattac 5581 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga 5641 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc 5701 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa 5761 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc 5821 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc 5881 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc 5941 gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg 6001 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct 6061 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg 6121 agtcggatct ccctttgggc cgcctccccg cctggtacct ttaagaccaa tgacttacaa 6181 ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca 6241 ctcccaacga agataagatc tgctttttgc ttgtactggg tctctctggt tagaccagat 6301 ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt 6361 gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc 6421 cctcagaccc ttttagtcag tgtggaaaat ctctagcagt agtagttcat gtcatcttat 6481 tattcagtat ttataacttg caaagaaatg aatatcagag agtgagagga acttgtttat 6541 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt 6601 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg 6661 gctctagcta tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc 6721 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg 6781 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctaga cttttgcaga 6841 gaccaaattc gtaatcatgt catagctgtt tcctgtgtga aattgttatc cgctcacaat 6901 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag 6961 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg 7021 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc 7081 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc 7141 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa 7201 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt 7261 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg 7321 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg 7381 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag 7441 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc 7501 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa 7561 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg 7621 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc 7681 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac 7741 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg 7801 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt 7861 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt 7921 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa 7981 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga 8041 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt 8101 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg 8161 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga 8221 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga 8281 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg 8341 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc 8401 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc 8461 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca 8521 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac 8581 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg 8641 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc 8701 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg 8761 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac 8821 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat 8881 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata 8941 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa 9001 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg 9061 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat 9121 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg 9181 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga 9241 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag 9301 aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc 9361 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt 9421 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa 9481 gctg //