LOCUS V038695 8836 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V038695 VERSION V038695 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..3690 /label="Pdcd10(NM_019745)" /note="Pdcd10(NM_019745)" /gene="Pdcd10" CDS 3697..3762 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3806..4017 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4030..4298 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 4325..4924 /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 4931..5519 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5593..5826 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 5898..6019 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6038..6173 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6211..6227) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6235..6251 /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(6259..6289) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6304..6325 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6613..7201) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7372..8232) /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(8233..8337) /gene="bla" /label="AmpR promoter" primer_bind 8811..8827 /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 tgaccgtcag atcgcctgga gacgccatcc acgctgtttt gacctccata gaagattcta 2281 gagctagcga attcagccac 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 aggaggggga agcggtggag gcggttctct cgagatgagg 3061 atgacaatgg aagagatgaa gaatgaagct gagaccactt ccatggtttc tatgcctctc 3121 tatgcagtca tgtatcctgt gtttaatgag ctagaacgag taaatttgtc tgcagctcag 3181 actctgagag ccgctttcat taaggctgaa aaggaaaatc caggcctcac acaagacatc 3241 attatgaaaa tattagagaa aaaaagtgta gaagttaact tcaccgagtc cctccttcgt 3301 atggcagctg atgacgtaga agagtacatg attgagcggc cagagccaga attccaggac 3361 ctaaatgaaa aggcacgggc acttaaacag attctcagta agattcctga tgagatcaat 3421 gacagagtga ggtttctgca gacaatcaag gatatagcta gtgcaataaa agaactactt 3481 gatacagtga ataatgtctt caaaaaatat cagtaccaga accgcagggc acttgaacac 3541 caaaagaaag aatttgtaaa atactccaaa agtttcagtg acactctgaa gacttatttt 3601 aaagatggca aggcaataaa tgtgttcata agtgccaacc ggctaattca tcaaaccaac 3661 ttgatacttc agaccttcaa aactgtggcc ggatccgact acaaagacca tgacggtgat 3721 tataaagatc atgacatcga ctacaaggat gacgatgaca agtagcggcc gcgaaggatc 3781 tgcgatcgct ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag 3841 ttggggggag gggtcggcaa ttgaacgggt gcctagagaa ggtggcgcgg ggtaaactgg 3901 gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata 3961 agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacagctg 4021 aagcttcgag gggctcgcat ctctccttca cgcgcccgcc gccctacctg aggccgccat 4081 ccacgccggt tgagtcgcgt tctgccgcct cccgcctgtg gtgcctcctg aactgcgtcc 4141 gccgtctagg taagtttaaa gctcaggtcg agaccgggcc tttgtccggc gctcccttgg 4201 agcctaccta gactcagccg gctctccacg ctttgcctga ccctgcttgc tcaactctac 4261 gtctttgttt cgttttctgt tctgcgccgt tacagatcca agctgtgacc ggcgcctacg 4321 ctagatgacc gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tcccccgggc 4381 cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgaccc 4441 ggaccgccac atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct 4501 cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc 4561 ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag 4621 cggttcccgg ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa 4681 ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct 4741 gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt 4801 cctggagacc tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac 4861 cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc 4921 ctgagtcgac aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa 4981 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat 5041 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta 5101 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc 5161 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt 5221 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg 5281 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc 5341 ttggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc 5401 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct 5461 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc 5521 tggtaccttt aagaccaatg acttacaagg cagctgtaga tcttagccac tttttaaaag 5581 aaaagggggg actggaaggg ctaattcact cccaacgaaa ataagatctg ctttttgctt 5641 gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc taactaggga 5701 acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc 5761 tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct 5821 ctagcagtag tagttcatgt catcttatta ttcagtattt ataacttgca aagaaatgaa 5881 tatcagagag tgagaggaac ttgtttattg cagcttataa tggttacaaa taaagcaata 5941 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca 6001 aactcatcaa tgtatcttat catgtctggc tctagctatc ccgcccctaa ctccgcccag 6061 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc 6121 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctagactt 6181 ttgcagagac ggcccaaatt cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta 6241 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc 6301 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg 6361 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg 6421 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg 6481 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa 6541 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc 6601 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc 6661 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag 6721 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct 6781 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta 6841 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc 6901 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc 6961 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt 7021 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct 7081 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc 7141 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca 7201 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta 7261 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa 7321 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg 7381 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg 7441 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc 7501 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc 7561 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa 7621 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc 7681 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg 7741 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc 7801 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat 7861 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg 7921 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc 7981 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg 8041 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat 8101 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg 8161 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg 8221 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct 8281 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac 8341 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta 8401 taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa 8461 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag 8521 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta 8581 tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag 8641 atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg 8701 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc 8761 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac 8821 ggccagtgcc aagctg //