LOCUS V031766 9466 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V031766 VERSION V031766 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..3492 /label="CERS6(NM_001256126)" /note="CERS6(NM_001256126)" /gene="CERS6" CDS 3499..3564 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3608..3819 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3832..4100 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4129..4138 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4159..4821 /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 4891..4944 /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 4945..5544 /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 5545..6133 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6207..6440 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6512..6633 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6673..6808 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6841..6857) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6865..6881 /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(6889..6919) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6934..6955 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7243..7831) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8002..8862) /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(8863..8967) /gene="bla" /label="AmpR promoter" primer_bind 9441..9457 /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 cagggatctt agcctggttc 2341 tggaacgaga ggttttggct cccgcacaat gtcacctggg cggacctgaa gaacacggag 2401 gaggccacct tcccgcaggc tgaggacctc tatctcgctt ttcccctggc cttctgtatc 2461 ttcatggtgc ggctcatctt cgagagattt gtagccaaac cgtgcgccat agccctcaac 2521 attcaggcca atggaccaca aattgctccg cccaatgcca ttctggaaaa ggtcttcact 2581 gcaattacaa agcatcctga tgaaaagaga ttggaaggcc tctccaagca actggactgg 2641 gatgttcgaa gcattcagcg ctggtttcga caaagacgca atcaggagaa gccaagcacg 2701 ctgacgaggt tctgtgagag catgtggaga ttttcatttt acctttatgt atttacctac 2761 ggagtcagat tcctgaaaaa gaccccctgg ttgtggaata cgaggcattg ctggtacaac 2821 tacccctatc agccactcac aactgacctt cactactatt acatcctgga gctgtcgttt 2881 tattggtctt tgatgttttc tcagttcact gatatcaaaa gaaaggactt tggcattatg 2941 ttcctgcacc accttgtatc tattttcttg attacctttt catatgtcaa caatatggcc 3001 cgagtaggaa cgctggtcct ttgtcttcat gattcagctg atgctcttct ggaggctgcc 3061 aaaatggcaa attatgccaa gtttcagaaa atgtgtgatc tcctgtttgt tatgtttgcc 3121 gtggttttta tcaccacacg actgggtata tttcctctct gggtgttaaa taccacatta 3181 tttgaaagct gggagatcgt tggaccttac ccttcctggt gggtttttaa cctactgcta 3241 ttgctagtac aagggttgaa ctgcttctgg tcttacttga ttgtgaaaat agcttgcaaa 3301 gctgtttcaa gaggcaaggc tgggaagtgg aaccctttac atgtgtccaa ggatgatcga 3361 agtgatattg agtctagctc agatgaggag gactcagaac ctccgggaaa gaatccccac 3421 actgcgacaa ccaccaatgg gaccagtggt accaacgggt atctcctgac tggctcctgc 3481 tccatggatg atggatccga ctacaaagac catgacggtg attataaaga tcatgacatc 3541 gactacaagg atgacgatga caagtagcgg ccgcgaagga tctgcgatcg ctccggtgcc 3601 cgtcagtggg cagagcgcac atcgcccaca gtccccgaga agttgggggg aggggtcggc 3661 aattgaacgg gtgcctagag aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac 3721 tggctccgcc tttttcccga gggtggggga gaaccgtata taagtgcagt agtcgccgtg 3781 aacgttcttt ttcgcaacgg gtttgccgcc agaacacagc tgaagcttcg aggggctcgc 3841 atctctcctt cacgcgcccg ccgccctacc tgaggccgcc atccacgccg gttgagtcgc 3901 gttctgccgc ctcccgcctg tggtgcctcc tgaactgcgt ccgccgtcta ggtaagttta 3961 aagctcaggt cgagaccggg cctttgtccg gcgctccctt ggagcctacc tagactcagc 4021 cggctctcca cgctttgcct gaccctgctt gctcaactct acgtctttgt ttcgttttct 4081 gttctgcgcc gttacagatc caagctgtga ccggcgccta cgctagacgc caccatggag 4141 agcgacgaga gcggcctgcc cgccatggag atcgagtgcc gcatcaccgg caccctgaac 4201 ggcgtggagt tcgagctggt gggcggcgga gagggcaccc ccaagcaggg ccgcatgacc 4261 aacaagatga agagcaccaa aggcgccctg accttcagcc cctacctgct gagccacgtg 4321 atgggctacg gcttctacca cttcggcacc taccccagcg gctacgagaa ccccttcctg 4381 cacgccatca acaacggcgg ctacaccaac acccgcatcg agaagtacga ggacggcggc 4441 gtgctgcacg tgagcttcag ctaccgctac gaggccggcc gcgtgatcgg cgacttcaag 4501 gtggtgggca ccggcttccc cgaggacagc gtgatcttca ccgacaagat catccgcagc 4561 aacgccaccg tggagcacct gcaccccatg ggcgataacg tgctggtggg cagcttcgcc 4621 cgcaccttca gcctgcgcga cggcggctac tacagcttcg tggtggacag ccacatgcac 4681 ttcaagagcg ccatccaccc cagcatcctg cagaacgggg gccccatgtt cgccttccgc 4741 cgcgtggagg agctgcacag caacaccgag ctgggcatcg tggagtacca gcacgccttc 4801 aagaccccca tcgccttcgc cagatcccgc gctcagtcgt ccaattctgc cgtggacggc 4861 accgccggac ccggctccac cggatctcgc gagggcagag gaagtcttct aacatgcggt 4921 gacgtggagg agaatcccgg ccctatgacc gagtacaagc ccacggtgcg cctcgccacc 4981 cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc 5041 acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct gcaagaactc 5101 ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg 5161 gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc 5221 ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggaaggcctc 5281 ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc 5341 gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag 5401 cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag 5461 cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc 5521 atgacccgca agcccggtgc ctgaaatcaa cctctggatt acaaaatttg tgaaagattg 5581 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct 5641 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg 5701 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact 5761 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc 5821 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc 5881 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa 5941 tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc 6001 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg 6061 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg 6121 gccgcctccc cgcctggtac ctttaagacc aatgacttac aaggcagctg tagatcttag 6181 ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac gaagataaga 6241 tctgcttttt gcttgtactg ggtctctctg gttagaccag atctgagcct gggagctctc 6301 tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt 6361 agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc 6421 agtgtggaaa atctctagca gtagtagttc atgtcatctt attattcagt atttataact 6481 tgcaaagaaa tgaatatcag agagtgagag gaacttgttt attgcagctt ataatggtta 6541 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag 6601 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggctctagc tatcccgccc 6661 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc 6721 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag 6781 aagtagtgag gaggcttttt tggaggccta gacttttgca gagaccaaat tcgtaatcat 6841 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc 6901 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc 6961 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat 7021 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac 7081 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt 7141 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca 7201 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc 7261 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact 7321 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct 7381 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag 7441 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca 7501 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa 7561 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc 7621 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag 7681 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg 7741 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca 7801 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc 7861 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag 7921 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata 7981 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat 8041 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg 8101 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc 8161 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc 8221 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc 8281 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc 8341 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc 8401 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa 8461 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat 8521 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata 8581 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca 8641 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag 8701 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc 8761 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc 8821 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata 8881 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta 8941 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta 9001 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg 9061 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt 9121 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg 9181 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt 9241 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg 9301 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct 9361 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg 9421 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctg //