LOCUS V029090 9864 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V029090 VERSION V029090 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 1494..1538 /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 1801..1916 /note="central polypurine tract and central termination sequence of HIV-1 (lacking the first T)" /label="cPPT/CTS" enhancer 1974..2277 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 2278..2481 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 2588..3934 /label="Rbfox2(NM_053104)" /note="Rbfox2(NM_053104)" /gene="Rbfox2" CDS 3959..4030 /codon_start=1 /product="three tandem FLAG® epitope tags" /transl_table=1 /translation="DYKDDDDKDYKDDDDKDYKDDDDK" /label="3xFLAG" promoter 4084..4295 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4308..4576 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 4604..5296 /codon_start=1 /note="mammalian codon-optimized" /product="Zoanthus green fluorescent protein" /transl_table=1 /translation="MAQSKHGLTKEMTMKYRMEGCVDGHKFVITGEGIGYPFKGKQAIN LCVVEGGPLPFAEDILSAAFMYGNRVFTEYPQDIVDYFKNSCPAGYTWDRSFLFEDGAV CICNADITVSVEENCMYHESKFYGVNFPADGPVMKKMTDNWEPSCEKIIPVPKQGILKG DVSMYLLLKDGGRLRCQFDTVYKAKSVPRKMPDWHFIQHKLTREDRSDAKNQKWHLTEH AIASGSALP" /label="ZsGreen1" CDS 5306..5359 /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 5360..5959 /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 5960..6547 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6621..6854 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6926..7047 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 7066..7201 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(7239..7255) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7263..7279 /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(7287..7317) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7332..7353 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7641..8229) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8400..9260) /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(9261..9365) /gene="bla" /label="AmpR promoter" primer_bind 9839..9855 /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 gggcgcagcg 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 acagatttgg aatcacacga cctggatgga gtgggacaga gaaattaaca 1441 attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa gaaaagaatg 1501 aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt aacataacaa 1561 attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta ggtttaagaa 1621 tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca ccattatcgt 1681 ttcagaccca cctcccaacc ccgaggggac ccgacaggcc cgaaggaata gaagaagaag 1741 gtggagagag agacagagac agatccattc gattagtgaa cggatctcga cggtatcgcc 1801 tttaaaagaa aaggggggat tggggggtac agtgcagggg aaagaatagt agacataata 1861 gcaacagaca tacaaactaa agaattacaa aaacaaatta caaaaattca aaattttcgg 1921 gtttattaca gggacagcag agatccagtt tatcgataag cttgggagtt ccgcgttaca 1981 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca 2041 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg 2101 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg 2161 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc 2221 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg 2281 atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca 2341 agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt 2401 ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg 2461 gaggtctata taagcagagc tcgtttagtg aaccgtcaga tcgcctggag acgccatcca 2521 cgctgttttg acctccatag aagacaccga ctctactaga ggatctattt ccggtgaatt 2581 cgccaccatg gcggaaggcg gccaggcgca gcagcagccg cctcagctcg ggcccggcgc 2641 tgccgcccgg ggcatgaagc gggagtcgga ggtggagttg ccggtgcccg gagcgggggc 2701 agacggaccc gagcccggcc tgagcaagcg gccacgcaca gaggaggcgg ccgacggcgg 2761 gatgcagaac gagccactga ccccgggtta tcatggattc ccagcaaggg atggccaggg 2821 taaccaggag ccaacaacaa ctcctgacgc aatggttcag ccttttacta ccatcccgtt 2881 cccaccacct ccacaaaatg gaattcccac agaatatgga gtgccacaca ctcaggacta 2941 tgccggccag accagtgaac ataacctgac cctctacggg agtacgcagc ctcatggaga 3001 acagagtagc aattcaccca gcaaccagaa tggatctctc acgcagacag aaggtggagc 3061 acagacagac ggacaacagt cacagacaca aagtagtgaa aattcagaga gtaaatctac 3121 gcccaagcga ctacatgtct ctaatattcc cttccgcttc cgagaccctg acctccggca 3181 gatgtttggg cagtttggca aaatcctaga tgtggaaata atctttaatg agcgcggctc 3241 caagggattc gggttcgtaa ctttcgagaa tagtgctgat gcagacaggg ccagggagaa 3301 attgcacggc accgtggtag agggccgtaa aatcgaggtg aataatgcaa cagcacgggt 3361 catgaccaac aagaagatgg tcacgccata tgcaaatggc tggaagttaa gcccagtagt 3421 tggagctgtg tatggtcctg agttatatgc agcatccagc tttcaagctg atgtgtccct 3481 aggcaatgag gcggctgtgc ccttgtcagg aagaggaggc atcaacactt acatccctct 3541 aatcagtctc cctttagttc ctggcttccc ttacccaact gcagccacca cggcagctgc 3601 tttccgagga gcccatctga ggggcagagg gcggacagtg tatggtgcag tccgagcggt 3661 acctccaaca gccatccccg cctatccagg tgtggtttac caggacggat tttacggtgc 3721 tgacctctat ggtggatatg cagcctacag atatgcacag cctgctactg caaccgcagc 3781 cacagctgct gcagccgctg cagccgctta cagcgacggt tacggcaggg tgtacacagc 3841 tgacccctac catgccctcg cccctgccgc cagctatgga gttggcgctg tggcgagttt 3901 gtaccgaggt ggctacagcc gatttgcccc ctacggatcc tcggtaccaa gcttaagtga 3961 ctacaaggat gacgatgaca aggattacaa agacgacgat gataaggact ataaggatga 4021 tgacgacaaa taaagatcca tcgatactag taaggatctg cgatcgctcc ggtgcccgtc 4081 agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt 4141 gaacgggtgc ctagagaagg tggcgcgggg taaactggga aagtgatgtc gtgtactggc 4201 tccgcctttt tcccgagggt gggggagaac cgtatataag tgcagtagtc gccgtgaacg 4261 ttctttttcg caacgggttt gccgccagaa cacagctgaa gcttcgaggg gctcgcatct 4321 ctccttcacg cgcccgccgc cctacctgag gccgccatcc acgccggttg agtcgcgttc 4381 tgccgcctcc cgcctgtggt gcctcctgaa ctgcgtccgc cgtctaggta agtttaaagc 4441 tcaggtcgag accgggcctt tgtccggcgc tcccttggag cctacctaga ctcagccggc 4501 tctccacgct ttgcctgacc ctgcttgctc aactctacgt ctttgtttcg ttttctgttc 4561 tgcgccgtta cagatccaag ctgtgaccgg cgcctactct agcatggccc agtccaagca 4621 cggcctgacc aaggagatga ccatgaagta ccgcatggag ggctgcgtgg acggccacaa 4681 gttcgtgatc accggcgagg gcatcggcta ccccttcaag ggcaagcagg ccatcaacct 4741 gtgcgtggtg gagggcggcc ccttgccctt cgccgaggac atcttgtccg ccgccttcat 4801 gtacggcaac cgcgtgttca ccgagtaccc ccaggacatc gtcgactact tcaagaactc 4861 ctgccccgcc ggctacacct gggaccgctc cttcctgttc gaggacggcg ccgtgtgcat 4921 ctgcaacgcc gacatcaccg tgagcgtgga ggagaactgc atgtaccacg agtccaagtt 4981 ctacggcgtg aacttccccg ccgacggccc cgtgatgaag aagatgaccg acaactggga 5041 gccctcctgc gagaagatca tccccgtgcc caagcagggc atcttgaagg gcgacgtgag 5101 catgtacctg ctgctgaagg acggtggccg cttgcgctgc cagttcgaca ccgtgtacaa 5161 ggccaagtcc gtgccccgca agatgcccga ctggcacttc atccagcaca agctgacccg 5221 cgaggaccgc agcgacgcca agaaccagaa gtggcacctg accgagcacg ccatcgcctc 5281 cggctccgcc ttgcccgcgg ccgctgaggg cagaggaagt cttctaacat gcggtgacgt 5341 ggaggagaat cccggcccta tgaccgagta caagcccacg gtgcgcctcg ccacccgcga 5401 cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg 5461 ccacaccgtc gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct 5521 cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc 5581 ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg 5641 catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc 5701 gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca 5761 ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc 5821 cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct 5881 cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac 5941 ccgcaagccc ggtgcctgaa atcaacctct ggattacaaa atttgtgaaa gattgactgg 6001 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta 6061 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct 6121 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt 6181 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac 6241 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg 6301 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac 6361 gtcctttcca tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg 6421 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct 6481 gcggcctctt ccgcgtctcc gccttcgccc tcagacgagt cggatctccc tttggccgcc 6541 tccccgcctg gtacctttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt 6601 tttaaaagaa aaggggggac tggaagggct aattcactcc caacgaaaat aagatctgct 6661 ttttgcttgt actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta 6721 actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg 6781 tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg 6841 gaaaatctct agcagtagta gttcatgtca tcttattatt cagtatttat aacttgcaaa 6901 gaaatgaata tcagagagtg agaggaactt gtttattgca gcttataatg gttacaaata 6961 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg 7021 tttgtccaaa ctcatcaatg tatcttatca tgtctggctc tagctatccc gcccctaact 7081 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag 7141 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc 7201 ctagactttt gcagagacgg cccaaattcg taatcatggt catagctgtt tcctgtgtga 7261 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc 7321 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc 7381 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc 7441 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt 7501 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca 7561 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa 7621 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat 7681 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc 7741 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc 7801 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt 7861 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac 7921 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg 7981 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca 8041 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc 8101 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa 8161 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa 8221 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac 8281 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta 8341 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt 8401 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata 8461 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc 8521 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac 8581 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag 8641 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac 8701 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc 8761 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg 8821 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc 8881 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct 8941 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc 9001 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc 9061 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc 9121 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc 9181 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca 9241 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt 9301 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt 9361 ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca 9421 ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac 9481 ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat 9541 gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg 9601 cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata 9661 ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc attcgccatt caggctgcgc 9721 aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg 9781 ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt 9841 aaaacgacgg ccagtgccaa gctg //