LOCUS V045448 8846 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V045448 VERSION V045448 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..3699 /label="HMGB1(NM_001313893)" /note="HMGB1(NM_001313893)" /gene="HMGB1" CDS 3706..3771 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3816..4027 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4040..4308 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 4335..4934 /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 4941..5529 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5603..5836 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 5908..6029 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6048..6183 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6221..6237) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6245..6261 /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(6269..6299) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6314..6335 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6623..7211) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7382..8242) /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(8243..8347) /gene="bla" /label="AmpR promoter" primer_bind 8821..8837 /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 atccatgggc 3061 aaaggagatc ctaagaagcc gagaggcaaa atgtcatcat atgcattttt tgtgcaaact 3121 tgtcgggagg agcataagaa gaagcaccca gatgcttcag tcaacttctc agagttttct 3181 aagaagtgct cagagaggtg gaagaccatg tctgctaaag agaaaggaaa atttgaagat 3241 atggcaaaag cggacaaggc ccgttatgaa agagaaatga aaacctatat ccctcccaaa 3301 ggggagacaa aaaagaagtt caaggatccc aatgcaccca agaggcctcc ttcggccttc 3361 ttcctcttct gctctgagta tcgcccaaaa atcaaaggag aacatcctgg cctgtccatt 3421 ggtgatgttg cgaagaaact gggagagatg tggaataaca ctgctgcaga tgacaagcag 3481 ccttatgaaa agaaggctgc gaagctgaag gaaaaatacg aaaaggatat tgctgcatat 3541 cgagctaaag gaaagcctga tgcagcaaaa aagggagttg tcaaggctga aaaaagcaag 3601 aaaaagaagg aagaggagga agatgaggaa gatgaagagg atgaggagga ggaggaagat 3661 gaagaagatg aagatgaaga agaagatgat gatgatgaag gatccgacta caaagaccat 3721 gacggtgatt ataaagatca tgacatcgac tacaaggatg acgatgacaa gtaggcggcc 3781 gcgaaggatc tgcgatcgct ccggtgcccg tcagtgggca gagcgcacat cgcccacagt 3841 ccccgagaag ttggggggag gggtcggcaa ttgaacgggt gcctagagaa ggtggcgcgg 3901 ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg gtgggggaga 3961 accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag 4021 aacacagctg aagcttcgag gggctcgcat ctctccttca cgcgcccgcc gccctacctg 4081 aggccgccat ccacgccggt tgagtcgcgt tctgccgcct cccgcctgtg gtgcctcctg 4141 aactgcgtcc gccgtctagg taagtttaaa gctcaggtcg agaccgggcc tttgtccggc 4201 gctcccttgg agcctaccta gactcagccg gctctccacg ctttgcctga ccctgcttgc 4261 tcaactctac gtctttgttt cgttttctgt tctgcgccgt tacagatcca agctgtgacc 4321 ggcgcctacg ctagatgacc gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg 4381 tcccccgggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca 4441 ccgtcgaccc ggaccgccac atcgagcggg tcaccgagct gcaagaactc ttcctcacgc 4501 gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct 4561 ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg 4621 ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc 4681 accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg 4741 gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg 4801 tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag cggctcggct 4861 tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc atgacccgca 4921 agcccggtgc ctgagtcgac aatcaacctc tggattacaa aatttgtgaa agattgactg 4981 gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt 5041 atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc 5101 tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt 5161 ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga 5221 ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct 5281 gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat 5341 cgtcctttcc ttggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct 5401 gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc 5461 tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg 5521 cctccccgcc tggtaccttt aagaccaatg acttacaagg cagctgtaga tcttagccac 5581 tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaaa ataagatctg 5641 ctttttgctt gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc 5701 taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg 5761 tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg 5821 tggaaaatct ctagcagtag tagttcatgt catcttatta ttcagtattt ataacttgca 5881 aagaaatgaa tatcagagag tgagaggaac ttgtttattg cagcttataa tggttacaaa 5941 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt 6001 ggtttgtcca aactcatcaa tgtatcttat catgtctggc tctagctatc ccgcccctaa 6061 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag 6121 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag 6181 gcctagactt ttgcagagac ggcccaaatt cgtaatcatg gtcatagctg tttcctgtgt 6241 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag 6301 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt 6361 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag 6421 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg 6481 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat 6541 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta 6601 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa 6661 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc 6721 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt 6781 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca 6841 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg 6901 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat 6961 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta 7021 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct 7081 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac 7141 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa 7201 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa 7261 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt 7321 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca 7381 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca 7441 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc 7501 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa 7561 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc 7621 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca 7681 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat 7741 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag 7801 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac 7861 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt 7921 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt 7981 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc 8041 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat 8101 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca 8161 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga 8221 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg 8281 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg 8341 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga 8401 cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg 8461 acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg 8521 atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct 8581 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa 8641 taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc 8701 gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag 8761 ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt 8821 gtaaaacgac ggccagtgcc aagctg //