LOCUS V042854 9226 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V042854 VERSION V042854 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..4080 /label="GRN(NM_002087)" /note="GRN(NM_002087)" /gene="GRN" CDS 4087..4152 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 4196..4407 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4420..4688 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 4715..5314 /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 5321..5909 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5983..6216 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6288..6409 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6428..6563 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6601..6617) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6625..6641 /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(6649..6679) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6694..6715 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7003..7591) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7762..8622) /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(8623..8727) /gene="bla" /label="AmpR promoter" primer_bind 9201..9217 /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 catgtggacc ctggtgagct gggtggcctt aacagcaggg 2341 ctggtggctg gaacgcggtg cccagatggt cagttctgcc ctgtggcctg ctgcctggac 2401 cccggaggag ccagctacag ctgctgccgt ccccttctgg acaaatggcc cacaacactg 2461 agcaggcatc tgggtggccc ctgccaggtt gatgcccact gctctgccgg ccactcctgc 2521 atctttaccg tctcagggac ttccagttgc tgccccttcc cagaggccgt ggcatgcggg 2581 gatggccatc actgctgccc acggggcttc cactgcagtg cagacgggcg atcctgcttc 2641 caaagatcag gtaacaactc cgtgggtgcc atccagtgcc ctgatagtca gttcgaatgc 2701 ccggacttct ccacgtgctg tgttatggtc gatggctcct gggggtgctg ccccatgccc 2761 caggcttcct gctgtgaaga cagggtgcac tgctgtccgc acggtgcctt ctgcgacctg 2821 gttcacaccc gctgcatcac acccacgggc acccaccccc tggcaaagaa gctccctgcc 2881 cagaggacta acagggcagt ggccttgtcc agctcggtca tgtgtccgga cgcacggtcc 2941 cggtgccctg atggttctac ctgctgtgag ctgcccagtg ggaagtatgg ctgctgccca 3001 atgcccaacg ccacctgctg ctccgatcac ctgcactgct gcccccaaga cactgtgtgt 3061 gacctgatcc agagtaagtg cctctccaag gagaacgcta ccacggacct cctcactaag 3121 ctgcctgcgc acacagtggg ggatgtgaaa tgtgacatgg aggtgagctg cccagatggc 3181 tatacctgct gccgtctaca gtcgggggcc tggggctgct gcccttttac ccaggctgtg 3241 tgctgtgagg accacataca ctgctgtccc gcggggttta cgtgtgacac gcagaagggt 3301 acctgtgaac aggggcccca ccaggtgccc tggatggaga aggccccagc tcacctcagc 3361 ctgccagacc cacaagcctt gaagagagat gtcccctgtg ataatgtcag cagctgtccc 3421 tcctccgata cctgctgcca actcacgtct ggggagtggg gctgctgtcc aatcccagag 3481 gctgtctgct gctcggacca ccagcactgc tgcccccagg gctacacgtg tgtagctgag 3541 gggcagtgtc agcgaggaag cgagatcgtg gctggactgg agaagatgcc tgcccgccgg 3601 gcttccttat cccaccccag agacatcggc tgtgaccagc acaccagctg cccggtgggg 3661 cagacctgct gcccgagcct gggtgggagc tgggcctgct gccagttgcc ccatgctgtg 3721 tgctgcgagg atcgccagca ctgctgcccg gctggctaca cctgcaacgt gaaggctcga 3781 tcctgcgaga aggaagtggt ctctgcccag cctgccacct tcctggcccg tagccctcac 3841 gtgggtgtga aggacgtgga gtgtggggaa ggacacttct gccatgataa ccagacctgc 3901 tgccgagaca accgacaggg ctgggcctgc tgtccctacc gccagggcgt ctgttgtgct 3961 gatcggcgcc actgctgtcc tgctggcttc cgctgcgcag ccaggggtac caagtgtttg 4021 cgcagggagg ccccgcgctg ggacgcccct ttgagggacc cagccttgag acagctgctg 4081 ggatccgact acaaagacca tgacggtgat tataaagatc atgacatcga ctacaaggat 4141 gacgatgaca agtagcggcc gcgaaggatc tgcgatcgct ccggtgcccg tcagtgggca 4201 gagcgcacat cgcccacagt ccccgagaag ttggggggag gggtcggcaa ttgaacgggt 4261 gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt 4321 tttcccgagg gtgggggaga accgtatata agtgcagtag tcgccgtgaa cgttcttttt 4381 cgcaacgggt ttgccgccag aacacagctg aagcttcgag gggctcgcat ctctccttca 4441 cgcgcccgcc gccctacctg aggccgccat ccacgccggt tgagtcgcgt tctgccgcct 4501 cccgcctgtg gtgcctcctg aactgcgtcc gccgtctagg taagtttaaa gctcaggtcg 4561 agaccgggcc tttgtccggc gctcccttgg agcctaccta gactcagccg gctctccacg 4621 ctttgcctga ccctgcttgc tcaactctac gtctttgttt cgttttctgt tctgcgccgt 4681 tacagatcca agctgtgacc ggcgcctacg ctagatgacc gagtacaagc ccacggtgcg 4741 cctcgccacc cgcgacgacg tcccccgggc cgtacgcacc ctcgccgccg cgttcgccga 4801 ctaccccgcc acgcgccaca ccgtcgaccc ggaccgccac atcgagcggg tcaccgagct 4861 gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga 4921 cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc 4981 cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat 5041 ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg 5101 cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga 5161 ggcggccgag cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc 5221 cttctacgag cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg 5281 cacctggtgc atgacccgca agcccggtgc ctgagtcgac aatcaacctc tggattacaa 5341 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata 5401 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc 5461 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg 5521 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac 5581 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat 5641 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt 5701 ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat 5761 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc 5821 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag 5881 tcggatctcc ctttgggccg cctccccgcc tggtaccttt aagaccaatg acttacaagg 5941 cagctgtaga tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact 6001 cccaacgaaa ataagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct 6061 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc 6121 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc 6181 tcagaccctt ttagtcagtg tggaaaatct ctagcagtag tagttcatgt catcttatta 6241 ttcagtattt ataacttgca aagaaatgaa tatcagagag tgagaggaac ttgtttattg 6301 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt 6361 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctggc 6421 tctagctatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact 6481 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta 6541 gtgaggaggc ttttttggag gcctagactt ttgcagagac ggcccaaatt cgtaatcatg 6601 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc 6661 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc 6721 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat 6781 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac 6841 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt 6901 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca 6961 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc 7021 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact 7081 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct 7141 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag 7201 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca 7261 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa 7321 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc 7381 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag 7441 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg 7501 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca 7561 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc 7621 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag 7681 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata 7741 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat 7801 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg 7861 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc 7921 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc 7981 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc 8041 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc 8101 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc 8161 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa 8221 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat 8281 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata 8341 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca 8401 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag 8461 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc 8521 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc 8581 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata 8641 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta 8701 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta 8761 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg 8821 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt 8881 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg 8941 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt 9001 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg 9061 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct 9121 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg 9181 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctg //