LOCUS V023701 9024 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V023701 VERSION V023701 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..3878 /label="FTO(NM_001363894)" /note="FTO(NM_001363894)" /gene="FTO" CDS 3885..3950 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3994..4205 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4218..4486 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 4513..5112 /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 5119..5707 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5781..6014 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6086..6207 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6226..6361 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6399..6415) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6423..6439 /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(6447..6477) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6492..6513 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6801..7389) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7560..8420) /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(8421..8525) /gene="bla" /label="AmpR promoter" primer_bind 8999..9015 /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 atgaagcgca ccccgactgc cgaggaacga gagcgcgaag 2341 ctaagaaact gaggcttctt gaagagcttg aagacacttg gctcccttat ctgaccccca 2401 aagatgatga attctatcag cagtggcagc tgaaatatcc taaactaatt ctccgagaag 2461 ccagcagtgt atctgaggag ctccataaag aggttcaaga agcctttctc acactgcaca 2521 agcatggctg cttatttcgg gacctggtta ggatccaagg caaagatctg ctcactccgg 2581 tatctcgcat cctcattggt aatccaggct gcacctacaa gtacctgaac accaggctct 2641 ttacggtccc ctggccagtg aaagggtcta atataaaaca caccgaggct gaaatagccg 2701 ctgcttgtga gaccttcctc aagctcaatg actacctgca gatagaaacc atccaggctt 2761 tggaagaact tgctgccaaa gagaaggcta atgaggatgc tgtgccattg tgtatgtctg 2821 cagatttccc cagggttggg atgggttcat cctacaacgg acaagatgaa gtggacatta 2881 agagcagagc agcatacaac gtaactttgc tgaatttcat ggatcctcag aaaatgccat 2941 acctgaaaga ggaaccttat tttggcatgg ggaaaatggc agtgagctgg catcatgatg 3001 aaaatctggt ggacaggtca gcggtggcag tgtacagtta tagctgtgaa ggccctgaag 3061 aggaaagtga ggatgactct catctcgaag gcagggatcc tgatatttgg catgttggtt 3121 ttaagatctc atgggacata gagacacctg gtttggcgat accccttcac caaggagact 3181 gctatttcat gcttgatgat ctcaatgcca cccaccaaca ctgtgttttg gccggttcac 3241 aacctcggtt tagttccacc caccgagtgg cagagtgctc aacaggaacc ttggattata 3301 ttttacaacg ctgtcagttg gctctgcaga atgtctgtga cgatgtggac aatgatgatg 3361 tctctttgaa atcctttgag cctgcagttt tgaaacaagg agaagaaatt cataatgagg 3421 tcgagtttga gtggctgagg cagttttggt ttcaaggcaa tcgatacaga aagtgcactg 3481 actggtggtg tcaacccatg gctcaactgg aagcactgtg gaagaagatg gagggtgtga 3541 taagtgtgga gttatacaaa gagctaggac ttatgtctcc caggttgata aggcacaagg 3601 caacaaatgc tgtgcttcat gaagttaaaa gagaggggct ccccgtggaa caaaggaatg 3661 aaatcttgac tgccatcctt gcctcgctca ctgcacgcca gaacctgagg agagaatggc 3721 atgccaggtg ccagtcacga attgcccgaa cattacctgc tgatcagaag ccagaatgtc 3781 ggccatactg ggaaaaggat gatgcttcga tgcctctgcc gtttgacctc acagacatcg 3841 tttcagaact cagaggtcag cttctggaag caaaacccgg atccgactac aaagaccatg 3901 acggtgatta taaagatcat gacatcgact acaaggatga cgatgacaag tagcggccgc 3961 gaaggatctg cgatcgctcc ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc 4021 ccgagaagtt ggggggaggg gtcggcaatt gaacgggtgc ctagagaagg tggcgcgggg 4081 taaactggga aagtgatgtc gtgtactggc tccgcctttt tcccgagggt gggggagaac 4141 cgtatataag tgcagtagtc gccgtgaacg ttctttttcg caacgggttt gccgccagaa 4201 cacagctgaa gcttcgaggg gctcgcatct ctccttcacg cgcccgccgc cctacctgag 4261 gccgccatcc acgccggttg agtcgcgttc tgccgcctcc cgcctgtggt gcctcctgaa 4321 ctgcgtccgc cgtctaggta agtttaaagc tcaggtcgag accgggcctt tgtccggcgc 4381 tcccttggag cctacctaga ctcagccggc tctccacgct ttgcctgacc ctgcttgctc 4441 aactctacgt ctttgtttcg ttttctgttc tgcgccgtta cagatccaag ctgtgaccgg 4501 cgcctacgct agatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc 4561 ccccgggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc 4621 gtcgacccgg accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc 4681 gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg 4741 accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc 4801 gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac 4861 cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc 4921 aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg 4981 cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc 5041 accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag 5101 cccggtgcct gagtcgacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt 5161 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat 5221 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg 5281 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt 5341 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact 5401 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc 5461 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg 5521 tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc 5581 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg 5641 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc 5701 tccccgcctg gtacctttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt 5761 tttaaaagaa aaggggggac tggaagggct aattcactcc caacgaaaat aagatctgct 5821 ttttgcttgt actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta 5881 actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg 5941 tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg 6001 gaaaatctct agcagtagta gttcatgtca tcttattatt cagtatttat aacttgcaaa 6061 gaaatgaata tcagagagtg agaggaactt gtttattgca gcttataatg gttacaaata 6121 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg 6181 tttgtccaaa ctcatcaatg tatcttatca tgtctggctc tagctatccc gcccctaact 6241 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag 6301 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc 6361 ctagactttt gcagagacgg cccaaattcg taatcatggt catagctgtt tcctgtgtga 6421 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc 6481 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc 6541 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc 6601 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt 6661 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca 6721 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa 6781 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat 6841 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc 6901 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc 6961 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt 7021 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac 7081 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg 7141 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca 7201 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc 7261 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa 7321 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa 7381 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac 7441 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta 7501 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt 7561 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata 7621 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc 7681 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac 7741 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag 7801 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac 7861 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc 7921 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg 7981 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc 8041 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct 8101 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc 8161 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc 8221 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc 8281 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc 8341 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca 8401 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt 8461 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt 8521 ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca 8581 ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac 8641 ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat 8701 gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg 8761 cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata 8821 ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc attcgccatt caggctgcgc 8881 aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg 8941 ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt 9001 aaaacgacgg ccagtgccaa gctg //