LOCUS V028804 8224 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V028804 VERSION V028804 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..3054 /label="Fgf23(NM_022657)" /note="Fgf23(NM_022657)" /gene="Fgf23" CDS 3073..3102 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /transl_table=1 /translation="EQKLISEEDL" /label="Myc" CDS 3121..3144 /codon_start=1 /product="FLAG® epitope tag, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDDDDK" /label="FLAG" promoter 3194..3405 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3418..3686 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 3713..4312 /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 4319..4907 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 4981..5214 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 5286..5407 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 5426..5561 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(5599..5615) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5623..5639 /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(5647..5677) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5692..5713 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6001..6589) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6760..7620) /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(7621..7725) /gene="bla" /label="AmpR promoter" primer_bind 8199..8215 /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 catgctaggg acctgcctta gactcctggt gggcgtgctc 2341 tgcactgtct gcagcttggg cactgctaga gcctatccgg acacttcccc attgcttggc 2401 tccaactggg gaagcctgac ccacctgtac acggctacag ccaggaccag ctatcaccta 2461 cagatccata gggatggtca tgtagatggc accccccatc agaccatcta cagtgccctg 2521 atgattacat cagaggacgc cggctctgtg gtgataacag gagccatgac tcgaaggttc 2581 ctttgtatgg atctccacgg caacattttt ggatcgcttc acttcagccc agagaattgc 2641 aagttccgcc agtggacgct ggagaatggc tatgacgtct acttgtcgca gaagcatcac 2701 tacctggtga gcctgggccg cgccaagcgc atcttccagc cgggcaccaa cccgccgccc 2761 ttctcccagt tcctggctcg caggaacgag gtcccgctgc tgcatttcta cactgttcgc 2821 ccacggcgcc acacgcgcag cgccgaggac ccaccggagc gcgacccact gaacgtgctc 2881 aagccgcggc cccgcgccac gcctgtgcct gtatcctgct ctcgcgagct gccgagcgca 2941 gaggaaggtg gccccgcagc cagcgatcct ctgggggtgc tgcgcagagg ccgtggagat 3001 gctcgcgggg gcgcgggagg cgcggatagg tgtcgcccct ttcccaggtt cgtcacgcgt 3061 acgcggccgc tcgagcagaa actcatctca gaagaggatc tggcagcaaa tgatatcctg 3121 gattacaagg atgacgacga taagtaggat ccgcggccgc gaaggatctg cgatcgctcc 3181 ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg 3241 gtcggcaatt gaacgggtgc ctagagaagg tggcgcgggg taaactggga aagtgatgtc 3301 gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag tgcagtagtc 3361 gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacagctgaa gcttcgaggg 3421 gctcgcatct ctccttcacg cgcccgccgc cctacctgag gccgccatcc acgccggttg 3481 agtcgcgttc tgccgcctcc cgcctgtggt gcctcctgaa ctgcgtccgc cgtctaggta 3541 agtttaaagc tcaggtcgag accgggcctt tgtccggcgc tcccttggag cctacctaga 3601 ctcagccggc tctccacgct ttgcctgacc ctgcttgctc aactctacgt ctttgtttcg 3661 ttttctgttc tgcgccgtta cagatccaag ctgtgaccgg cgcctacgct agatgaccga 3721 gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc ccccgggccg tacgcaccct 3781 cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgacccgg accgccacat 3841 cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa 3901 ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga 3961 agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct 4021 ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg 4081 gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt 4141 cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc 4201 cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga 4261 ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gagtcgacaa 4321 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc 4381 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat 4441 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg 4501 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg 4561 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat 4621 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt 4681 gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc 4741 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa 4801 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg 4861 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg gtacctttaa 4921 gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac 4981 tggaagggct aattcactcc caacgaaaat aagatctgct ttttgcttgt actgggtctc 5041 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta 5101 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact 5161 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtagta 5221 gttcatgtca tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg 5281 agaggaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt 5341 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg 5401 tatcttatca tgtctggctc tagctatccc gcccctaact ccgcccagtt ccgcccattc 5461 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc 5521 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctagactttt gcagagacgg 5581 cccaaattcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat 5641 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag 5701 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg 5761 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc 5821 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc 5881 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa 5941 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt 6001 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg 6061 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg 6121 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag 6181 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc 6241 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa 6301 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg 6361 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc 6421 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac 6481 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg 6541 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt 6601 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt 6661 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa 6721 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga 6781 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt 6841 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg 6901 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga 6961 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga 7021 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg 7081 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc 7141 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc 7201 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca 7261 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac 7321 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg 7381 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc 7441 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg 7501 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac 7561 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat 7621 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata 7681 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa 7741 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg 7801 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat 7861 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg 7921 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga 7981 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag 8041 aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc 8101 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt 8161 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa 8221 gctg //