LOCUS V029354 8604 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V029354 VERSION V029354 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" regulatory 2288..2297 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 2297..2362 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" CDS 2369..3529 /label="GMPPB(NM_013334)" /note="GMPPB(NM_013334)" /gene="GMPPB" promoter 3574..3785 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3798..4066 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 4093..4692 /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 4699..5287 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5361..5594 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 5666..5787 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 5806..5941 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(5979..5995) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6003..6019 /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(6027..6057) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6072..6093 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6381..6969) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7140..8000) /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(8001..8105) /gene="bla" /label="AmpR promoter" primer_bind 8579..8595 /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 agagctagcc accatggact acaaagacca tgacggtgat tataaagatc atgacatcga 2341 ctacaaggat gacgatgaca aggaattcat gaaggcactg atcttagtgg ggggctatgg 2401 gacgcggcta cggccgctga cgctgagcac cccgaagcca ctggtggact tctgcaataa 2461 gcccatcttg ctgcaccaag tggaggcgct agccgcggca ggcgtggacc acgtgatcct 2521 ggccgtgagc tacatgtcgc aggtgctgga gaaggaaatg aaggcacagg agcagaggct 2581 gggaatccga atctccatgt cccatgaaga ggagcctttg gggacagctg ggcccctggc 2641 gctggcccgt gacctactct ctgagactgc agaccctttc ttcgtcctca acagtgacgt 2701 gatctgcgat ttccccttcc aagccatggt gcagttccac cggcaccatg gccaggaggg 2761 ctccatcctg gtgaccaagg tggaggaacc ctccaagtac ggtgtggtgg tgtgtgaggc 2821 tgacacaggc cgcattcacc ggttcgtgga gaagccacag gtgtttgtgt ccaataagat 2881 caacgcaggc atgtacatcc tgagccctgc agtgctgcag cgcatccagc tgcagcctac 2941 gtccattgag aaggaggtct tccccattat ggccaaggag gggcagctat atgccatgga 3001 gttacagggc ttctggatgg acattgggca gcccaaggac ttcctcactg gcatgtgcct 3061 cttcctgcag tcactgaggc agaagcagcc tgagcggctg tgctcaggcc ctggcattgt 3121 gggcaacgtg ctggtggacc caagtgcccg catcggccag aactgcagca ttggccccaa 3181 tgtgagcctg ggacctggcg tggtggtcga agatggtgtg tgtatccggc ggtgcacggt 3241 gctgcgggat gcccggatcc gttcccattc ctggcttgag tcctgcattg tgggctggcg 3301 ctgccgcgtg ggtcagtggg taagcctgtg ggctgggctg ggtggggaga ggggcgggga 3361 gtgtgcctgc ctccctgaca aggcctatcc tctcctggag gtacgcatgg agaacgtgac 3421 agtgctgggt gaggacgtca tagttaatga tgagctctac ctcaacggag ccagcgtgct 3481 gccccacaag tctattggcg agtcagtgcc agagcctcgt atcatcatgt gagcggccgc 3541 gaaggatctg cgatcgctcc ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc 3601 ccgagaagtt ggggggaggg gtcggcaatt gaacgggtgc ctagagaagg tggcgcgggg 3661 taaactggga aagtgatgtc gtgtactggc tccgcctttt tcccgagggt gggggagaac 3721 cgtatataag tgcagtagtc gccgtgaacg ttctttttcg caacgggttt gccgccagaa 3781 cacagctgaa gcttcgaggg gctcgcatct ctccttcacg cgcccgccgc cctacctgag 3841 gccgccatcc acgccggttg agtcgcgttc tgccgcctcc cgcctgtggt gcctcctgaa 3901 ctgcgtccgc cgtctaggta agtttaaagc tcaggtcgag accgggcctt tgtccggcgc 3961 tcccttggag cctacctaga ctcagccggc tctccacgct ttgcctgacc ctgcttgctc 4021 aactctacgt ctttgtttcg ttttctgttc tgcgccgtta cagatccaag ctgtgaccgg 4081 cgcctacgct agatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc 4141 ccccgggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc 4201 gtcgacccgg accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc 4261 gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg 4321 accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc 4381 gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac 4441 cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc 4501 aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg 4561 cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc 4621 accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag 4681 cccggtgcct gagtcgacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt 4741 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat 4801 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg 4861 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt 4921 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact 4981 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc 5041 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg 5101 tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc 5161 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg 5221 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc 5281 tccccgcctg gtacctttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt 5341 tttaaaagaa aaggggggac tggaagggct aattcactcc caacgaaaat aagatctgct 5401 ttttgcttgt actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta 5461 actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg 5521 tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg 5581 gaaaatctct agcagtagta gttcatgtca tcttattatt cagtatttat aacttgcaaa 5641 gaaatgaata tcagagagtg agaggaactt gtttattgca gcttataatg gttacaaata 5701 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg 5761 tttgtccaaa ctcatcaatg tatcttatca tgtctggctc tagctatccc gcccctaact 5821 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag 5881 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc 5941 ctagactttt gcagagacgg cccaaattcg taatcatggt catagctgtt tcctgtgtga 6001 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc 6061 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc 6121 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc 6181 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt 6241 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca 6301 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa 6361 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat 6421 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc 6481 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc 6541 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt 6601 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac 6661 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg 6721 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca 6781 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc 6841 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa 6901 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa 6961 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac 7021 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta 7081 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt 7141 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata 7201 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc 7261 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac 7321 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag 7381 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac 7441 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc 7501 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg 7561 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc 7621 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct 7681 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc 7741 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc 7801 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc 7861 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc 7921 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca 7981 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt 8041 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt 8101 ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca 8161 ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac 8221 ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat 8281 gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg 8341 cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata 8401 ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc attcgccatt caggctgcgc 8461 aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg 8521 ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt 8581 aaaacgacgg ccagtgccaa gctg //