LOCUS V020178 8806 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V020178 VERSION V020178 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 1798..1915 /note="central polypurine tract and central termination sequence of HIV-1" /label="cPPT/CTS" promoter 2005..2208 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 2297..2848 /label="Tnfaip8l2(NM_027206)" /note="Tnfaip8l2(NM_027206)" /gene="Tnfaip8l2" CDS 2855..2920 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 2965..3176 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3189..3457 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 3486..3495 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 3516..4178 /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 4248..4301 /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 4302..4901 /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 4902..5489 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5563..5796 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 5868..5989 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6008..6143 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6181..6197) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6205..6221 /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(6229..6259) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6274..6295 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6583..7171) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7342..8202) /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(8203..8307) /gene="bla" /label="AmpR promoter" primer_bind 8781..8797 /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 ggttaacttt 1801 taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc 1861 aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa attttatcga 1921 tactagtatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac 1981 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga 2041 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg 2101 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg 2161 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctcg tttagtgaac 2221 cgtcagatcg cctggagacg ccatccacgc tgttttgacc tccatagaag attctagagc 2281 tagcgaattc gccaccatgg agtccttcag ctcaaagagt ctggcactac aagcggagaa 2341 gaagctgctg agtaaaatgg ctggtcggtc cgtggcgcat ctctttatcg acgagaccag 2401 cagcgaggtg ctagacgagc tttaccgcgt gtccaaagaa tacacgcaca gccggcccaa 2461 ggcacagcgg gtgatcaaag acctcatcaa ggtagcggtt aaagtggctg tgctgcaccg 2521 cagtggctgc tttggccctg gggagctggc tctggctaca cgatttcgtc agaagctacg 2581 gcagggcgcc atgaccgcac ttagcttcgg tgaggtggac ttcacctttg aggctgccgt 2641 gctagcaggt ctgctcgtcg agtgccggga cattctgctg gagctggtgg agcaccacct 2701 cacacccaag tcacatgacc gcatcaggca cgtgtttgat cactactctg accccgacct 2761 gctggctgcc ctctatgggc ctgacttcac tcagcacctt ggcaagatct gtgatgggct 2821 ccggaagctg ctggacgagg gcaagctcgg atccgactac aaagaccatg acggtgatta 2881 taaagatcat gacatcgact acaaggatga cgatgacaag tgagcggccg cgaaggatct 2941 gcgatcgctc cggtgcccgt cagtgggcag agcgcacatc gcccacagtc cccgagaagt 3001 tggggggagg ggtcggcaat tgaacgggtg cctagagaag gtggcgcggg gtaaactggg 3061 aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa ccgtatataa 3121 gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt tgccgccaga acacagctga 3181 agcttcgagg ggctcgcatc tctccttcac gcgcccgccg ccctacctga ggccgccatc 3241 cacgccggtt gagtcgcgtt ctgccgcctc ccgcctgtgg tgcctcctga actgcgtccg 3301 ccgtctaggt aagtttaaag ctcaggtcga gaccgggcct ttgtccggcg ctcccttgga 3361 gcctacctag actcagccgg ctctccacgc tttgcctgac cctgcttgct caactctacg 3421 tctttgtttc gttttctgtt ctgcgccgtt acagatccaa gctgtgaccg gcgcctacgc 3481 tagacgccac catggagagc gacgagagcg gcctgcccgc catggagatc gagtgccgca 3541 tcaccggcac cctgaacggc gtggagttcg agctggtggg cggcggagag ggcaccccca 3601 agcagggccg catgaccaac aagatgaaga gcaccaaagg cgccctgacc ttcagcccct 3661 acctgctgag ccacgtgatg ggctacggct tctaccactt cggcacctac cccagcggct 3721 acgagaaccc cttcctgcac gccatcaaca acggcggcta caccaacacc cgcatcgaga 3781 agtacgagga cggcggcgtg ctgcacgtga gcttcagcta ccgctacgag gccggccgcg 3841 tgatcggcga cttcaaggtg gtgggcaccg gcttccccga ggacagcgtg atcttcaccg 3901 acaagatcat ccgcagcaac gccaccgtgg agcacctgca ccccatgggc gataacgtgc 3961 tggtgggcag cttcgcccgc accttcagcc tgcgcgacgg cggctactac agcttcgtgg 4021 tggacagcca catgcacttc aagagcgcca tccaccccag catcctgcag aacgggggcc 4081 ccatgttcgc cttccgccgc gtggaggagc tgcacagcaa caccgagctg ggcatcgtgg 4141 agtaccagca cgccttcaag acccccatcg ccttcgccag atcccgcgct cagtcgtcca 4201 attctgccgt ggacggcacc gccggacccg gctccaccgg atctcgcgag ggcagaggaa 4261 gtcttctaac atgcggtgac gtggaggaga atcccggccc tatgaccgag tacaagccca 4321 cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt 4381 tcgccgacta ccccgccacg cgccacaccg tcgatccgga ccgccacatc gagcgggtca 4441 ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg 4501 cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg 4561 tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc 4621 aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca 4681 ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg 4741 gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca 4801 acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag 4861 gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg aaatcaacct ctggattaca 4921 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat 4981 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct 5041 ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac 5101 gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca 5161 cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca 5221 tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg 5281 tggtgttgtc ggggaagctg acgtcctttc catggctgct cgcctgtgtt gccacctgga 5341 ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt 5401 cccgcggcct gctgccggct ctgcggcctc ttccgcgtct ccgccttcgc cctcagacga 5461 gtcggatctc cctttggccg cctccccgcc tggtaccttt aagaccaatg acttacaagg 5521 cagctgtaga tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact 5581 cccaacgaaa ataagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct 5641 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc 5701 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc 5761 tcagaccctt ttagtcagtg tggaaaatct ctagcagtag tagttcatgt catcttatta 5821 ttcagtattt ataacttgca aagaaatgaa tatcagagag tgagaggaac ttgtttattg 5881 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt 5941 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctggc 6001 tctagctatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact 6061 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta 6121 gtgaggaggc ttttttggag gcctagactt ttgcagagac ggcccaaatt cgtaatcatg 6181 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc 6241 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc 6301 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat 6361 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac 6421 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt 6481 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca 6541 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc 6601 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact 6661 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct 6721 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag 6781 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca 6841 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa 6901 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc 6961 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag 7021 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg 7081 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca 7141 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc 7201 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag 7261 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata 7321 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat 7381 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg 7441 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc 7501 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc 7561 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc 7621 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc 7681 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc 7741 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa 7801 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat 7861 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata 7921 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca 7981 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag 8041 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc 8101 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc 8161 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata 8221 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta 8281 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta 8341 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg 8401 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt 8461 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg 8521 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt 8581 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg 8641 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct 8701 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg 8761 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctg //