LOCUS V030063 9910 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V030063 VERSION V030063 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..3936 /label="CNOT2(NM_001199303)" /note="CNOT2(NM_001199303)" /gene="CNOT2" CDS 3943..4008 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 4052..4263 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4276..4544 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4573..4582 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4603..5265 /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 5335..5388 /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 5389..5988 /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 5989..6577 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6651..6884 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6956..7077 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 7117..7252 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(7285..7301) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7309..7325 /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(7333..7363) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7378..7399 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7687..8275) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8446..9306) /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(9307..9411) /gene="bla" /label="AmpR promoter" primer_bind 9885..9901 /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 tgaggactga tggacataca 2341 ttatctgaga aaagaaacta ccaggtgaca aacagcatgt ttggtgcttc aagaaagaag 2401 tttgtagagg gggtcgacag tgactaccat gacgaaaaca tgtactacag ccagtcttct 2461 atgtttccac atcggtcaga aaaagatatg ctggcatcac catctacatc aggtcagctg 2521 tctcagtttg gggcaagttt atacgggcaa caaagtgcac taggccttcc aatgaggggg 2581 atgagcaaca atacccctca gttaaatcgc agcttatcac aaggcactca gttaccgagc 2641 cacgtcacgc caacaacagg ggtaccaaca atgtcacttc acacgcctcc atctccaagc 2701 aggggtattt tgcctatgaa tcctaggaat atgatgaacc actcccaggt tggtcagggc 2761 attggaattc ctagcaggac aaatagcatg agcagttcag ggttaggtag ccccaacaga 2821 agctcgccaa gcataatatg tatgccaaag cagcagcctt ctcgacagcc ttttactgtg 2881 aacagtatgt ctggatttgg aatgaacagg aatcaggcat ttggaatgaa taactcctta 2941 tcaagtaaca tttttaatgg aacagacgga agtgaaaatg tgacaggatt ggacctttca 3001 gatttcccag cattagcaga ccgaaacagg agggaaggaa gtggtaaccc aactccatta 3061 ataaacccct tggctggaag agctccttat gttggaatgg taacaaaacc agcaaatgaa 3121 caatcccagg acttctcaat acacaatgaa gattttccag cattaccagg ctccagctat 3181 aaagatccaa catcaagtaa tgatgacagt aaatctaatt tgaatacatc tggcaagaca 3241 acttcaagta cagatggacc caaattccct ggagataaaa gttcaacaac acaaaataat 3301 aaccagcaga aaaaagggat ccaggtgtta cctgatggtc gggttactaa cattcctcaa 3361 gggatggtga cggaccaatt tggaatgatt ggcctgttaa catttatcag ggcagcagag 3421 acagacccag gaatggtaca tcttgcatta ggaagtgact taacaacatt aggcctcaat 3481 ctgaactctc ctgaaaatct ctaccccaaa tttgcgtcac cctgggcatc ttcaccttgt 3541 cgacctcaag acatagactt ccatgttcca tctgagtact taacgaacat tcacattagg 3601 gataagctgg ctgcaataaa acttggccga tatggtgaag accttctctt ctatctctat 3661 tacatgaatg gaggagacgt attacaactt ttagctgcag tggagctttt taaccgtgat 3721 tggagatacc acaaagaaga acgagtatgg attaccaggg caccaggcat ggagccaaca 3781 atgaaaacca atacctatga gaggggaaca tattacttct ttgactgtct taactggagg 3841 aaagtagcta aggagttcca tctggaatat gacaaattag aagaacggcc tcacctgcca 3901 tccaccttca actacaaccc tgctcagcaa gccttcggat ccgactacaa agaccatgac 3961 ggtgattata aagatcatga catcgactac aaggatgacg atgacaagta gcggccgcga 4021 aggatctgcg atcgctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc 4081 gagaagttgg ggggaggggt cggcaattga acgggtgcct agagaaggtg gcgcggggta 4141 aactgggaaa gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg 4201 tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca 4261 cagctgaagc ttcgaggggc tcgcatctct ccttcacgcg cccgccgccc tacctgaggc 4321 cgccatccac gccggttgag tcgcgttctg ccgcctcccg cctgtggtgc ctcctgaact 4381 gcgtccgccg tctaggtaag tttaaagctc aggtcgagac cgggcctttg tccggcgctc 4441 ccttggagcc tacctagact cagccggctc tccacgcttt gcctgaccct gcttgctcaa 4501 ctctacgtct ttgtttcgtt ttctgttctg cgccgttaca gatccaagct gtgaccggcg 4561 cctacgctag acgccaccat ggagagcgac gagagcggcc tgcccgccat ggagatcgag 4621 tgccgcatca ccggcaccct gaacggcgtg gagttcgagc tggtgggcgg cggagagggc 4681 acccccaagc agggccgcat gaccaacaag atgaagagca ccaaaggcgc cctgaccttc 4741 agcccctacc tgctgagcca cgtgatgggc tacggcttct accacttcgg cacctacccc 4801 agcggctacg agaacccctt cctgcacgcc atcaacaacg gcggctacac caacacccgc 4861 atcgagaagt acgaggacgg cggcgtgctg cacgtgagct tcagctaccg ctacgaggcc 4921 ggccgcgtga tcggcgactt caaggtggtg ggcaccggct tccccgagga cagcgtgatc 4981 ttcaccgaca agatcatccg cagcaacgcc accgtggagc acctgcaccc catgggcgat 5041 aacgtgctgg tgggcagctt cgcccgcacc ttcagcctgc gcgacggcgg ctactacagc 5101 ttcgtggtgg acagccacat gcacttcaag agcgccatcc accccagcat cctgcagaac 5161 gggggcccca tgttcgcctt ccgccgcgtg gaggagctgc acagcaacac cgagctgggc 5221 atcgtggagt accagcacgc cttcaagacc cccatcgcct tcgccagatc ccgcgctcag 5281 tcgtccaatt ctgccgtgga cggcaccgcc ggacccggct ccaccggatc tcgcgagggc 5341 agaggaagtc ttctaacatg cggtgacgtg gaggagaatc ccggccctat gaccgagtac 5401 aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg caccctcgcc 5461 gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg ccacatcgag 5521 cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg 5581 tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg 5641 ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc 5701 gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc 5761 ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg 5821 ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg 5881 ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg 5941 cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgaaa tcaacctctg 6001 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta 6061 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt 6121 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc 6181 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt 6241 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg 6301 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac 6361 aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc 6421 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac 6481 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct 6541 cagacgagtc ggatctccct ttgggccgcc tccccgcctg gtacctttaa gaccaatgac 6601 ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct 6661 aattcactcc caacgaagat aagatctgct ttttgcttgt actgggtctc tctggttaga 6721 ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata 6781 aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta 6841 gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtagta gttcatgtca 6901 tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggaactt 6961 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa 7021 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca 7081 tgtctggctc tagctatccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt 7141 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc 7201 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctagacttt 7261 tgcagagacc aaattcgtaa tcatgtcata gctgtttcct gtgtgaaatt gttatccgct 7321 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg 7381 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct 7441 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg 7501 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc 7561 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg 7621 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct 7681 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca 7741 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct 7801 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc 7861 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt 7921 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc 7981 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc 8041 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg 8101 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc 8161 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag 8221 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga 8281 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat 8341 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag 8401 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat 8461 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc 8521 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat 8581 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag 8641 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg 8701 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc 8761 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca 8821 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg 8881 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc 8941 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta 9001 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc 9061 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg 9121 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc 9181 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc 9241 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat 9301 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag 9361 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc 9421 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa 9481 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg 9541 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca 9601 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc 9661 atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt 9721 aaggagaaaa taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg 9781 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag 9841 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag 9901 tgccaagctg //