LOCUS V046618 9611 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V046618 VERSION V046618 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" promoter 2010..2213 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 2302..3018 /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" CDS 3055..4443 /label="Ruvbl2(NM_011304)" /note="Ruvbl2(NM_011304)" /gene="Ruvbl2" CDS 4450..4476 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" CDS 4480..4506 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" CDS 4510..4536 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" promoter 4581..4792 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4805..5073 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 5100..5699 /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 5706..6294 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6368..6601 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6673..6794 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6813..6948 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6986..7002) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7010..7026 /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(7034..7064) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7079..7100 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7388..7976) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8147..9007) /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(9008..9112) /gene="bla" /label="AmpR promoter" primer_bind 9586..9602 /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 gtattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca 1981 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc 2041 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga 2101 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat 2161 tgacgcaaat gggcggtagg cgtgtacggt gggaggttta tataagcaga gctcgtttag 2221 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagattct 2281 agagctagcg aattcgccac catggtgagc aagggcgagg agctgttcac cggggtggtg 2341 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag 2401 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag 2461 ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc 2521 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac 2581 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg 2641 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag 2701 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc 2761 atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag 2821 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc 2881 gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac 2941 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc 3001 atggacgagc tgtacaaggg cggcggaggg agtggaggag ggggaagtgg atccatggca 3061 accgtggcag ccaccaccaa agtccctgag atccgagatg tgacaagaat cgagcgaatc 3121 ggcgctcact cccacatccg ggggctggga ctggatgatg ccctggagcc aagacaggct 3181 tcccagggaa tggtggggca gctagctgcc cggcgtgcag ctggcgtggt gctggagatg 3241 atccgagaag ggaagattgc agggcgggcc gtcctcattg caggccagcc aggcactggg 3301 aagacagcca ttgccatggg catggcccag gcactgggcc ccgacacacc attcacagcc 3361 atcgcaggca gtgagatctt ttctctggag atgagcaaga ctgaggctct aacgcaagcc 3421 ttccgccgtt ccattggtgt ccggatcaag gaggagacag agatcattga aggggaggtg 3481 gtagagatcc agattgaccg gccagccaca ggcacgggat ccaaggtggg caaactgacc 3541 ctcaagacca cggagatgga gaccatctac gacctgggta ccaagatgat cgagtccctg 3601 accaaggaca aggtccaggc cggagacgtc atcaccattg acaaggccac cggcaagatc 3661 tccaagctgg gccgctcttt cacacgtgca cgggactacg acgccatggg ctcccagacc 3721 aagtttgtgc agtgcccaga cggagagctg cagaaacgca aggaggtggt gcacaccgtg 3781 tccctccatg agattgacgt catcaactcc cgtactcagg gcttcctggc cctcttctca 3841 ggagacacag gggagatcaa gtcagaagtc cgagaacaga tcaatgcaaa ggtggctgag 3901 tggagagagg agggcaaggc agagatcatt cctggggtgc tgttcatcga cgaggtccac 3961 atgctggaca ttgagagttt ctctttcctc aaccgggccc tggagagcga catggcgcct 4021 gtcctcatca tggccaccaa ccgagggatc acccggatcc gaggcaccag ctaccagagt 4081 ccccacggca tccccatcga cctgctggac cgcttgctca ttgtatcaac gtcaccctac 4141 agtgagaagg ataccaagca gattctgcgc atccgctgtg aggaggaaga tgtggagatg 4201 agtgaggacg cctacacagt gctgacccgc attgggctcg agacctccct gcgctatgcc 4261 atccagctca tcacagcggc cagcctggtg tgccggaaac gaaagggcac cgaggtgcag 4321 gtcgatgaca ttaagcgtgt atactccctc ttcctggatg aatcccgctc cacacagtac 4381 atgaaggagt accaagatgc cttcctcttc aatgagctca aaggcgaaac aatggacacc 4441 tccctcgagt acccatacga cgtacctgat tatgcaactt atccttatga tgtacctgat 4501 tatgctagct atccttatga tgtcccagac tacgcatagg cggccgcgaa ggatctgcga 4561 tcgctccggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg 4621 gggaggggtc ggcaattgaa cgggtgccta gagaaggtgg cgcggggtaa actgggaaag 4681 tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc 4741 agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac agctgaagct 4801 tcgaggggct cgcatctctc cttcacgcgc ccgccgccct acctgaggcc gccatccacg 4861 ccggttgagt cgcgttctgc cgcctcccgc ctgtggtgcc tcctgaactg cgtccgccgt 4921 ctaggtaagt ttaaagctca ggtcgagacc gggcctttgt ccggcgctcc cttggagcct 4981 acctagactc agccggctct ccacgctttg cctgaccctg cttgctcaac tctacgtctt 5041 tgtttcgttt tctgttctgc gccgttacag atccaagctg tgaccggcgc ctacgctaga 5101 tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc cgggccgtac 5161 gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gacccggacc 5221 gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca 5281 tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga 5341 gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt 5401 cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc 5461 ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca 5521 gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg 5581 agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg 5641 acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgag 5701 tcgacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg 5761 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt 5821 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg 5881 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc 5941 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc 6001 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc 6061 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc 6121 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg 6181 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc 6241 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggta 6301 cctttaagac caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag 6361 gggggactgg aagggctaat tcactcccaa cgaaaataag atctgctttt tgcttgtact 6421 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca 6481 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg 6541 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc 6601 agtagtagtt catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca 6661 gagagtgaga ggaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc 6721 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc 6781 atcaatgtat cttatcatgt ctggctctag ctatcccgcc cctaactccg cccagttccg 6841 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct 6901 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta gacttttgca 6961 gagacggccc aaattcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc 7021 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat 7081 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc 7141 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg 7201 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag 7261 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag 7321 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc 7381 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc 7441 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc 7501 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt 7561 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg 7621 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat 7681 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag 7741 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt 7801 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc 7861 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta 7921 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag 7981 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga 8041 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa 8101 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa 8161 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc 8221 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga 8281 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa 8341 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt 8401 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg 8461 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc 8521 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg 8581 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag 8641 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt 8701 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt 8761 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac 8821 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac 8881 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag 8941 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa 9001 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga 9061 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc 9121 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa 9181 ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct 9241 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac 9301 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg 9361 catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg 9421 taaggagaaa ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag 9481 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa 9541 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca 9601 gtgccaagct g //