LOCUS V045189 9307 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V045189 VERSION V045189 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..1920 /note="central polypurine tract and central termination sequence of HIV-1" /label="cPPT/CTS" enhancer 1933..2236 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 2237..2440 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 2475..4526 /label="KBTBD7(NM_032138)" /note="KBTBD7(NM_032138)" /gene="KBTBD7" CDS 4533..4598 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" CDS 4608..4664 /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 porcine teschovirus-1 polyprotein" /transl_table=1 /translation="ATNFSLLKQAGDVEENPGP" /label="P2A" CDS 4677..5396 /codon_start=1 /note="mammalian codon-optimized" /product="the original enhanced GFP (Yang et al., 1996)" /transl_table=1 /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL TLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV LLEFVTAAGITLGMDELYK*" /label="EGFP" misc_feature 5403..5990 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6064..6297 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6369..6490 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6509..6644 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6682..6698) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6706..6722 /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(6730..6760) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6775..6796 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7084..7672) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7843..8703) /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(8704..8808) /gene="bla" /label="AmpR promoter" primer_bind 9282..9298 /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 attacaaaat tcaaaatttt 1921 atcgatacta gtcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga 1981 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt 2041 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt 2101 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca 2161 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt 2221 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt 2281 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca 2341 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg 2401 cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat 2461 ccgctagcgc caccatgcag tcccgggaag acgtcccgcg ctctcgccgc ctcgccagtc 2521 cccgtggtgg gaggcggccc aagaggattt ccaagccctc ggtttcggcc tttttcacgg 2581 gtccagagga gttaaaggac acggcccatt ctgcagccct gctggcacag ctcaagtcct 2641 tctacgacgc gcggctgctg tgtgatgtga ccatcgaggt ggtgacgcct ggcagcgggc 2701 ctggcacggg tcgcctcttt tcctgcaatc gcaacgtgct agcagctgcg tgtccctact 2761 tcaagagcat gttcacaggt ggcatgtacg agagccagca ggccagcgtg accatgcacg 2821 atgtggacgc cgagtccttc gaggtgttgg tcgactactg ctacacgggt cgtgtgtctc 2881 tcagtgaggc caatgtgcag cgcctgtacg cggcctccga catgctacag ctggaatatg 2941 tgcgggaagc ctgtgcctcc ttcttagccc gacgtcttga cctgaccaac tgcaccgcca 3001 tcctcaagtt tgcagacgcc ttcgaccatc acaagcttcg atctcaggcc cagtcctaca 3061 tagctcacaa cttcaagcag ctcagccgaa tgggttcaat tcgggaggag actctagcag 3121 atctaaccct ggcccagctg ctggctgtcc tacgcctgga tagtctggac atagagagtg 3181 agcggactgt atgccatgta gctgtgcagt ggctggaggc tgctgccaaa gagcggggtc 3241 ccagtgctgc agaagtcttc aagtgcgtgc gctggatgca cttcactgaa gaagatcagg 3301 actacttaga agggctgctg accaagccca tcgtgaagaa gtactgcctg gacgttattg 3361 aaggggccct gcagatgcgc tatggtgacc tgttgtacaa gtctctggtg ccagtgccaa 3421 acagcagcag cagcagtagc agcagcaact ctcttgtatc tgcagcagaa aatccacccc 3481 agagactggg tatgtgtgcc aaggagatgg tgatcttctt tggacatcct agagatccct 3541 ttctctgcta tgacccttac tcgggggaca tttacacaat gccatcccct ttgaccagct 3601 ttgctcacac taagactgtc acctcctcag ctgtctgtgt gtccccagac catgacatct 3661 atctagctgc tcagcccagg aaagacctct gggtgtataa accagctcag aatagttggc 3721 agcaacttgc agatcgcttg ctgtgtcgtg agggcatgga tgtggcatat ctcaatggct 3781 acatctacat tttgggggga cgagacccta ttactggagt taagttgaag gaagtggaat 3841 gctacagtgt tcagagaaac cagtgggcat tggtggctcc tgtccctcat tccttctatt 3901 cctttgaact catagtggtt cagaactatc tttatgctgt caacagtaag cgcatgcttt 3961 gctatgatcc tagccacaat atgtggctga actgtgcttc tcttaaacgt agtgactttc 4021 aggaagcatg tgtcttcaat gatgaaatct attgtatctg tgacatccca gtcatgaagg 4081 tctacaaccc agctagggga gaatggaggc ggattagtaa tattcctttg gattcagaga 4141 cccacaacta ccagattgtc aatcatgacc aaaagttgct tctcatcact tctacaaccc 4201 cacaatggaa aaagaaccga gtgacagtgt atgagtatga tactagggaa gatcagtgga 4261 ttaatatagg taccatgtta ggccttttgc agtttgactc tggctttatt tgcctttgtg 4321 ctcgtgttta tccttcctgc cttgaacctg gtcagagttt tattactgag gaagatgatg 4381 cacggagtga gtctagtact gaatgggact tagatggatt cagtgagctg gactctgagt 4441 caggaagttc aagttctttt tcagatgatg aagtctgggt gcaagtagca cctcagcgaa 4501 atgcacagga tcagcagggt tctttggtcg acgactacaa agaccatgac ggtgattata 4561 aagatcatga catcgactac aaggatgacg atgacaaggg aagcggagcc actaacttct 4621 ccctgttgaa acaagcaggg gatgtcgaag agaatcccgg gccagaattc gccaccatgg 4681 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg 4741 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca 4801 agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg 4861 tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc 4921 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca 4981 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga 5041 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc 5101 tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca 5161 tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc 5221 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc 5281 tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc 5341 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaaggat 5401 ccaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg 5461 ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc 5521 gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt 5581 tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca 5641 ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc 5701 ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc 5761 tgttgggcac tgacaattcc gtggtgttgt cggggaagct gacgtccttt ccatggctgc 5821 tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc 5881 tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc 5941 tccgccttcg ccctcagacg agtcggatct ccctttggcc gcctccccgc ctggtacctt 6001 taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg 6061 gactggaagg gctaattcac tcccaacgaa aataagatct gctttttgct tgtactgggt 6121 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc 6181 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg 6241 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta 6301 gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga 6361 gtgagaggaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa 6421 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca 6481 atgtatctta tcatgtctgg ctctagctat cccgccccta actccgccca gttccgccca 6541 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc 6601 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctagact tttgcagaga 6661 cggcccaaat tcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac 6721 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt 6781 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc 6841 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg 6901 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt 6961 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa 7021 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc 7081 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag 7141 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt 7201 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg 7261 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg 7321 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg 7381 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac 7441 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg 7501 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt 7561 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg 7621 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc 7681 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt 7741 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt 7801 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag 7861 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt 7921 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc 7981 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc 8041 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg 8101 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac 8161 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg 8221 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc 8281 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact 8341 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc 8401 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat 8461 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc 8521 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac 8581 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa 8641 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact 8701 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg 8761 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg 8821 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag 8881 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca 8941 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc 9001 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc 9061 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag 9121 gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg 9181 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg 9241 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtgc 9301 caagctg //