LOCUS V024696 8874 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V024696 VERSION V024696 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 2301..3728 /label="EGR2(NM_000399)" /note="EGR2(NM_000399)" /gene="EGR2" CDS 3735..3800 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3844..4055 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4068..4336 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 4363..4962 /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 4969..5557 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5631..5864 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 5936..6057 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6076..6211 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6249..6265) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6273..6289 /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(6297..6327) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6342..6363 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6651..7239) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7410..8270) /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(8271..8375) /gene="bla" /label="AmpR promoter" primer_bind 8849..8865 /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 attcgccacc atgatgaccg ccaaggccgt agacaaaatc ccagtaactc 2341 tcagtggttt tgtgcaccag ctgtctgaca acatctaccc ggtggaggac ctcgccgcca 2401 cgtcggtgac catctttccc aatgccgaac tgggaggccc ctttgaccag atgaacggag 2461 tggccggaga tggcatgatc aacattgaca tgactggaga gaagaggtcg ttggatctcc 2521 catatcccag cagctttgct cccgtctctg cacctagaaa ccagaccttc acttacatgg 2581 gcaagttctc cattgaccct cagtaccctg gtgccagctg ctacccagaa ggcataatca 2641 atattgtgag tgcaggcatc ttgcaagggg tcacttcccc agcttcaacc acagcctcat 2701 ccagcgtcac ctctgcctcc cccaacccac tggccacagg acccctgggt gtgtgcacca 2761 tgtcccagac ccagcctgac ctggaccacc tgtactctcc gccaccgcct cctcctcctt 2821 attctggctg tgcaggagac ctctaccagg acccttctgc gttcctgtca gcagccacca 2881 cctccacctc ttcctctctg gcctacccac cacctccttc ctatccatcc cccaagccag 2941 ccacggaccc aggtctcttc ccaatgatcc cagactatcc tggattcttt ccatctcagt 3001 gccagagaga cctacatggt acagctggcc cagaccgtaa gccctttccc tgcccactgg 3061 acaccctgcg ggtgccccct ccactcactc cactctctac aatccgtaac tttaccctgg 3121 ggggccccag tgctggggtg accggaccag gggccagtgg aggcagcgag ggaccccggc 3181 tgcctggtag cagctcagca gcagcagcag ccgccgccgc cgccgcctat aacccacacc 3241 acctgccact gcggcccatt ctgaggcctc gcaagtaccc caacagaccc agcaagacgc 3301 cggtgcacga gaggccctac ccgtgcccag cagaaggctg cgaccggcgg ttctcccgct 3361 ctgacgagct gacacggcac atccgaatcc acactgggca taagcccttc cagtgtcgga 3421 tctgcatgcg caacttcagc cgcagtgacc acctcaccac ccatatccgc acccacaccg 3481 gtgagaagcc cttcgcctgt gactactgtg gccgaaagtt tgcccggagt gatgagagga 3541 agcgccacac caagatccac ctgagacaga aagagcggaa aagcagtgcc ccctctgcat 3601 cggtgccagc cccctctaca gcctcctgct ctgggggcgt gcagcctggg ggtaccctgt 3661 gcagcagtaa cagcagcagt cttggcggag ggccgctcgc cccttgctcc tctcggaccc 3721 ggacacctgg atccgactac aaagaccatg acggtgatta taaagatcat gacatcgact 3781 acaaggatga cgatgacaag tagcggccgc gaaggatctg cgatcgctcc ggtgcccgtc 3841 agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt 3901 gaacgggtgc ctagagaagg tggcgcgggg taaactggga aagtgatgtc gtgtactggc 3961 tccgcctttt tcccgagggt gggggagaac cgtatataag tgcagtagtc gccgtgaacg 4021 ttctttttcg caacgggttt gccgccagaa cacagctgaa gcttcgaggg gctcgcatct 4081 ctccttcacg cgcccgccgc cctacctgag gccgccatcc acgccggttg agtcgcgttc 4141 tgccgcctcc cgcctgtggt gcctcctgaa ctgcgtccgc cgtctaggta agtttaaagc 4201 tcaggtcgag accgggcctt tgtccggcgc tcccttggag cctacctaga ctcagccggc 4261 tctccacgct ttgcctgacc ctgcttgctc aactctacgt ctttgtttcg ttttctgttc 4321 tgcgccgtta cagatccaag ctgtgaccgg cgcctacgct agatgaccga gtacaagccc 4381 acggtgcgcc tcgccacccg cgacgacgtc ccccgggccg tacgcaccct cgccgccgcg 4441 ttcgccgact accccgccac gcgccacacc gtcgacccgg accgccacat cgagcgggtc 4501 accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc 4561 gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg 4621 gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag 4681 caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc 4741 accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc 4801 ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc 4861 aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa 4921 ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gagtcgacaa tcaacctctg 4981 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta 5041 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt 5101 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc 5161 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt 5221 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg 5281 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac 5341 aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc 5401 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac 5461 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct 5521 cagacgagtc ggatctccct ttgggccgcc tccccgcctg gtacctttaa gaccaatgac 5581 ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct 5641 aattcactcc caacgaaaat aagatctgct ttttgcttgt actgggtctc tctggttaga 5701 ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata 5761 aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta 5821 gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtagta gttcatgtca 5881 tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggaactt 5941 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa 6001 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca 6061 tgtctggctc tagctatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat 6121 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc 6181 cagaagtagt gaggaggctt ttttggaggc ctagactttt gcagagacgg cccaaattcg 6241 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac 6301 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca 6361 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat 6421 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc 6481 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca 6541 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca 6601 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg 6661 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg 6721 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt 6781 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt 6841 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc 6901 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt 6961 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt 7021 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc 7081 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa 7141 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt 7201 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct 7261 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta 7321 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa 7381 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc 7441 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact 7501 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc 7561 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt 7621 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta 7681 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg 7741 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt 7801 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc 7861 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt 7921 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc 7981 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc 8041 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa 8101 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac 8161 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa 8221 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt 8281 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa 8341 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct 8401 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg 8461 ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg 8521 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg 8581 tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta 8641 ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc 8701 atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc 8761 tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta 8821 acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa gctg //