LOCUS V036965 10204 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V036965 VERSION V036965 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..4230 /label="ZNF143(NM_003442)" /note="ZNF143(NM_003442)" /gene="ZNF143" CDS 4237..4302 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 4346..4557 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4570..4838 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4867..4876 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4897..5559 /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 5629..5682 /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 5683..6282 /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 6283..6871 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6945..7178 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 7250..7371 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 7411..7546 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(7579..7595) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7603..7619 /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(7627..7657) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7672..7693 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7981..8569) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8740..9600) /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(9601..9705) /gene="bla" /label="AmpR promoter" primer_bind 10179..10195 /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 gccaccatgt tgttagccca aataaatcga 2341 gattctcagg gaatgacaga gtttcctgga ggagggatgg aggcgcaaca tgttacgctg 2401 tgcttgacag aggcagtcac cgtggcagat ggtgacaact tagaaaatat ggaaggcgta 2461 agcttgcaag cagtaacact tgcagatggt tctactgctt acatacaaca caattctaaa 2521 gatgcaaaac tcatagatgg ccaggtcatt cagttggaag atggttctgc ggcctatgtt 2581 caacatgtac ccatacctaa aagtacaggg gacagtttgc gtctagagga tggtcaagca 2641 gtacagttag aagatggtac cacagcattt attcaccaca cctccaaaga tagttatgac 2701 cagagtgcat tacaggcggt tcagctggaa gatggtacca cagcttatat ccaccatgca 2761 gtgcaagtcc cgcagtctga caccatcttg gcaattcagg ctgatgggac agtggcaggt 2821 ctgcacactg gggatgctac aattgaccct gacaccatca gtgctttgga acagtatgca 2881 gcaaaggtgt ccattgatgg aagtgaaagt gtagcaggta ctggaatgat tggagaaaat 2941 gagcaagaga aaaaaatgca gattgtttta caaggacatg ctacaagagt aactgctaaa 3001 tctcaacaga gtggagagaa ggcatttcga tgtgaatatg atggatgtgg aaaattatat 3061 acaacagctc atcatctcaa ggtccatgag aggtcacaca caggagatcg gccttatcag 3121 tgtgagcatg caggctgtgg gaaggcattt gcaacaggtt atggattaaa aagtcacgtc 3181 agaactcata caggagaaaa gccatatcgg tgttcggaag ataattgtac taaatctttc 3241 aaaacttcag gagatctaca gaaacacatc agaactcata caggagaaag gccctttaag 3301 tgtcccttcg aaggctgcgg tcggtccttt acaacatcaa atatcagaaa agtgcacgtt 3361 aggacacaca caggagaaag accttattac tgcacagagc caggatgtgg gagggcattt 3421 gccagtgcaa caaattataa aaaccatgtg aggatacaca caggagaaaa gccatatgtt 3481 tgtacagttc ctgggtgtga caaaaggttt acagaatatt ccagtttgta caaacatcat 3541 gttgtccaca ctcattccaa accttacaac tgtaaccact gtgggaagac atacaagcag 3601 atctccacgc tggccatgca caaacggaca gcccacaacg acactgagcc catcgaggag 3661 gagcaggaag ccttctttga gccgccccca ggtcaaggtg aagatgttct taaagggtcc 3721 cagattacgt atgttacagg tgtagaaggg gacgacgttg tttctacaca agtagccaca 3781 gtaacccaat ctggactgag tcaacaagtt acactcatat cccaggatgg gactcagcat 3841 gtcaacatat ctcaagctga catgcaggcc attggcaaca ccatcacaat ggtaacgcag 3901 gatggcacgc ccatcacagt ccccgcccat gatgcagtca tctcctcagc aggaacgcac 3961 tctgttgcta tggttactgc tgagggtaca gaaggggaac aggttgcaat tgtagctcaa 4021 gacttggcag cattccatac tgcctcatca gaaatggggc accagcagca tagccatcac 4081 ttagtaacca cagaaaccag acctctgacc ttagtagcaa catccaatgg cacccagatt 4141 gcagttcagc ttggagaaca gccatctctg gaagaagcca tcagaatagc gtctagaatc 4201 caacaaggag aaacgccagg gttggatgat ggatccgact acaaagacca tgacggtgat 4261 tataaagatc atgacatcga ctacaaggat gacgatgaca agtagcggcc gcgaaggatc 4321 tgcgatcgct ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag 4381 ttggggggag gggtcggcaa ttgaacgggt gcctagagaa ggtggcgcgg ggtaaactgg 4441 gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata 4501 agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacagctg 4561 aagcttcgag gggctcgcat ctctccttca cgcgcccgcc gccctacctg aggccgccat 4621 ccacgccggt tgagtcgcgt tctgccgcct cccgcctgtg gtgcctcctg aactgcgtcc 4681 gccgtctagg taagtttaaa gctcaggtcg agaccgggcc tttgtccggc gctcccttgg 4741 agcctaccta gactcagccg gctctccacg ctttgcctga ccctgcttgc tcaactctac 4801 gtctttgttt cgttttctgt tctgcgccgt tacagatcca agctgtgacc ggcgcctacg 4861 ctagacgcca ccatggagag cgacgagagc ggcctgcccg ccatggagat cgagtgccgc 4921 atcaccggca ccctgaacgg cgtggagttc gagctggtgg gcggcggaga gggcaccccc 4981 aagcagggcc gcatgaccaa caagatgaag agcaccaaag gcgccctgac cttcagcccc 5041 tacctgctga gccacgtgat gggctacggc ttctaccact tcggcaccta ccccagcggc 5101 tacgagaacc ccttcctgca cgccatcaac aacggcggct acaccaacac ccgcatcgag 5161 aagtacgagg acggcggcgt gctgcacgtg agcttcagct accgctacga ggccggccgc 5221 gtgatcggcg acttcaaggt ggtgggcacc ggcttccccg aggacagcgt gatcttcacc 5281 gacaagatca tccgcagcaa cgccaccgtg gagcacctgc accccatggg cgataacgtg 5341 ctggtgggca gcttcgcccg caccttcagc ctgcgcgacg gcggctacta cagcttcgtg 5401 gtggacagcc acatgcactt caagagcgcc atccacccca gcatcctgca gaacgggggc 5461 cccatgttcg ccttccgccg cgtggaggag ctgcacagca acaccgagct gggcatcgtg 5521 gagtaccagc acgccttcaa gacccccatc gccttcgcca gatcccgcgc tcagtcgtcc 5581 aattctgccg tggacggcac cgccggaccc ggctccaccg gatctcgcga gggcagagga 5641 agtcttctaa catgcggtga cgtggaggag aatcccggcc ctatgaccga gtacaagccc 5701 acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg 5761 ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc 5821 accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc 5881 gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg 5941 gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag 6001 caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc 6061 accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc 6121 ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc 6181 aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa 6241 ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gaaatcaacc tctggattac 6301 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga 6361 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc 6421 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa 6481 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc 6541 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc 6601 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc 6661 gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg 6721 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct 6781 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg 6841 agtcggatct ccctttgggc cgcctccccg cctggtacct ttaagaccaa tgacttacaa 6901 ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca 6961 ctcccaacga agataagatc tgctttttgc ttgtactggg tctctctggt tagaccagat 7021 ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt 7081 gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc 7141 cctcagaccc ttttagtcag tgtggaaaat ctctagcagt agtagttcat gtcatcttat 7201 tattcagtat ttataacttg caaagaaatg aatatcagag agtgagagga acttgtttat 7261 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt 7321 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg 7381 gctctagcta tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc 7441 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg 7501 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctaga cttttgcaga 7561 gaccaaattc gtaatcatgt catagctgtt tcctgtgtga aattgttatc cgctcacaat 7621 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag 7681 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg 7741 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc 7801 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc 7861 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa 7921 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt 7981 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg 8041 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg 8101 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag 8161 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc 8221 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa 8281 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg 8341 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc 8401 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac 8461 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg 8521 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt 8581 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt 8641 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa 8701 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga 8761 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt 8821 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg 8881 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga 8941 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga 9001 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg 9061 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc 9121 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc 9181 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca 9241 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac 9301 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg 9361 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc 9421 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg 9481 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac 9541 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat 9601 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata 9661 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa 9721 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg 9781 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat 9841 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg 9901 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga 9961 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag 10021 aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc 10081 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt 10141 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa 10201 gctg //