LOCUS V022278 9886 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V022278 VERSION V022278 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 2291..3928 /label="KAT5(NM_182710)" /note="KAT5(NM_182710)" /gene="KAT5" CDS 3935..4000 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 4045..4256 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4269..4537 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4566..4575 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4596..5258 /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 5328..5381 /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 5382..5981 /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 5982..6569 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6643..6876 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6948..7069 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 7088..7223 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(7261..7277) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7285..7301 /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(7309..7339) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7354..7375 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7663..8251) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8422..9282) /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(9283..9387) /gene="bla" /label="AmpR promoter" primer_bind 9861..9877 /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 tagcgccacc atggcggagg tggtgagtcc ggtgcccggg gcggggcgga gggagccagg 2341 ggaggtgggt agagcccgag gccccccagt agccgaccct ggcgtcgcgc tgtctcccca 2401 gggggagata atcgagggct gccgcctacc cgtgctgcgg cggaaccagg acaacgaaga 2461 tgagtggccc ctggccgaga tcctgagcgt gaaggacatc agtggccgga agcttttcta 2521 cgtccattac attgacttca acaaacgtct ggatgaatgg gtgacgcatg agcggctgga 2581 cctaaagaag atccagttcc ccaagaaaga ggccaagacc cccactaaga acggacttcc 2641 tgggtcccgt cctggctctc cagagagaga ggtgccggcc tcggcgcagg ccagcgggaa 2701 gaccttgcca atcccggtcc agatcacact ccgcttcaac ctgcccaagg agcgggaggc 2761 cattcccggt ggcgagcctg accagccgct ctcctccagc tcctgcctgc agcccaacca 2821 ccgctcaacg aaacggaagg tggaggtggt ttcaccagca actccagtgc ccagcgagac 2881 agccccggcc tcggtttttc cccagaatgg agccgcccgt agggcagtgg cagcccagcc 2941 aggacggaag cgaaaatcga attgtttggg cactgatgag gactcccagg acagctctga 3001 tggaataccg tcagcaccac gcatgactgg cagcctggtg tctgatcgaa gccacgacga 3061 catcgtcacc cggatgaaga acattgagtg cattgagctg ggccggcacc gcctcaagcc 3121 gtggtacttc tccccgtacc cacaggaact caccacattg cctgtcctct acctgtgcga 3181 gttctgcctc aagtacggcc gtagtctcaa gtgtcttcag cgtcatttga ccaagtgtga 3241 cctacgacat cctccaggca atgagattta ccgcaagggc accatctcct tctttgagat 3301 tgatggacgt aagaacaaga gttattccca gaacctgtgt cttttggcca agtgtttcct 3361 tgaccataag acactgtact atgacacaga ccctttcctc ttctacgtca tgacagagta 3421 tgactgtaag ggcttccaca tcgtgggcta cttctccaag gagaaagaat caacggaaga 3481 ctacaatgtg gcctgcatcc taaccctgcc tccctaccag cgccggggct acggcaagct 3541 gctgatcgag ttcagctatg aactctccaa agtggaaggg aaaacaggga cccctgagaa 3601 gcccctctca gaccttggcc tcctatccta tcgaagctac tggtcccaga ccatcctgga 3661 gatcctgatg gggctgaagt cggagagcgg ggagaggcca cagatcacca tcaatgagat 3721 tagtgaaatc accagcatca agaaggagga tgtcatctcc actctgcagt acctcaatct 3781 catcaactac tacaagggcc agtacatcct cacactgtca gaggacatcg tggatggcca 3841 tgagcgggcc atgctcaagc ggctcctgcg gatcgactcc aagtgtctgc acttcactcc 3901 caaggactgg agcaagaggg ggaagtgggg atccgactac aaagaccatg acggtgatta 3961 taaagatcat gacatcgact acaaggatga cgatgacaag tgagcggccg cgaaggatct 4021 gcgatcgctc cggtgcccgt cagtgggcag agcgcacatc gcccacagtc cccgagaagt 4081 tggggggagg ggtcggcaat tgaacgggtg cctagagaag gtggcgcggg gtaaactggg 4141 aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa ccgtatataa 4201 gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt tgccgccaga acacagctga 4261 agcttcgagg ggctcgcatc tctccttcac gcgcccgccg ccctacctga ggccgccatc 4321 cacgccggtt gagtcgcgtt ctgccgcctc ccgcctgtgg tgcctcctga actgcgtccg 4381 ccgtctaggt aagtttaaag ctcaggtcga gaccgggcct ttgtccggcg ctcccttgga 4441 gcctacctag actcagccgg ctctccacgc tttgcctgac cctgcttgct caactctacg 4501 tctttgtttc gttttctgtt ctgcgccgtt acagatccaa gctgtgaccg gcgcctacgc 4561 tagacgccac catggagagc gacgagagcg gcctgcccgc catggagatc gagtgccgca 4621 tcaccggcac cctgaacggc gtggagttcg agctggtggg cggcggagag ggcaccccca 4681 agcagggccg catgaccaac aagatgaaga gcaccaaagg cgccctgacc ttcagcccct 4741 acctgctgag ccacgtgatg ggctacggct tctaccactt cggcacctac cccagcggct 4801 acgagaaccc cttcctgcac gccatcaaca acggcggcta caccaacacc cgcatcgaga 4861 agtacgagga cggcggcgtg ctgcacgtga gcttcagcta ccgctacgag gccggccgcg 4921 tgatcggcga cttcaaggtg gtgggcaccg gcttccccga ggacagcgtg atcttcaccg 4981 acaagatcat ccgcagcaac gccaccgtgg agcacctgca ccccatgggc gataacgtgc 5041 tggtgggcag cttcgcccgc accttcagcc tgcgcgacgg cggctactac agcttcgtgg 5101 tggacagcca catgcacttc aagagcgcca tccaccccag catcctgcag aacgggggcc 5161 ccatgttcgc cttccgccgc gtggaggagc tgcacagcaa caccgagctg ggcatcgtgg 5221 agtaccagca cgccttcaag acccccatcg ccttcgccag atcccgcgct cagtcgtcca 5281 attctgccgt ggacggcacc gccggacccg gctccaccgg atctcgcgag ggcagaggaa 5341 gtcttctaac atgcggtgac gtggaggaga atcccggccc tatgaccgag tacaagccca 5401 cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt 5461 tcgccgacta ccccgccacg cgccacaccg tcgatccgga ccgccacatc gagcgggtca 5521 ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg 5581 cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg 5641 tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc 5701 aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca 5761 ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg 5821 gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca 5881 acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag 5941 gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg aaatcaacct ctggattaca 6001 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat 6061 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct 6121 ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac 6181 gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca 6241 cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca 6301 tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg 6361 tggtgttgtc ggggaagctg acgtcctttc catggctgct cgcctgtgtt gccacctgga 6421 ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt 6481 cccgcggcct gctgccggct ctgcggcctc ttccgcgtct ccgccttcgc cctcagacga 6541 gtcggatctc cctttggccg cctccccgcc tggtaccttt aagaccaatg acttacaagg 6601 cagctgtaga tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact 6661 cccaacgaaa ataagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct 6721 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc 6781 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc 6841 tcagaccctt ttagtcagtg tggaaaatct ctagcagtag tagttcatgt catcttatta 6901 ttcagtattt ataacttgca aagaaatgaa tatcagagag tgagaggaac ttgtttattg 6961 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt 7021 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctggc 7081 tctagctatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact 7141 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta 7201 gtgaggaggc ttttttggag gcctagactt ttgcagagac ggcccaaatt cgtaatcatg 7261 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc 7321 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc 7381 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat 7441 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac 7501 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt 7561 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca 7621 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc 7681 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact 7741 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct 7801 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag 7861 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca 7921 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa 7981 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc 8041 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag 8101 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg 8161 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca 8221 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc 8281 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag 8341 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata 8401 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat 8461 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg 8521 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc 8581 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc 8641 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc 8701 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc 8761 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc 8821 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa 8881 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat 8941 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata 9001 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca 9061 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag 9121 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc 9181 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc 9241 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata 9301 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta 9361 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta 9421 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg 9481 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt 9541 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg 9601 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt 9661 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg 9721 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct 9781 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg 9841 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctg //