LOCUS V042798 6998 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V042798 VERSION V042798 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers enhancer 202..581 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 582..785 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" promoter 830..848 /note="promoter for bacteriophage T7 RNA polymerase" /label="T7 promoter" CDS 911..931 /codon_start=1 /locus_tag="" /note="" /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /protein_id="" /transl_table=1 /translation="PKKKRKV" /label="SV40 NLS" CDS 932..952 /codon_start=1 /locus_tag="" /note="" /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /protein_id="" /transl_table=1 /translation="PKKKRKV" /label="SV40 NLS" CDS 953..973 /codon_start=1 /locus_tag="" /note="" /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /protein_id="" /transl_table=1 /translation="PKKKRKV" /label="SV40 NLS" CDS 974..2713 /label="METTL3(NM_019852)" /note="METTL3(NM_019852)" /gene="METTL3" polyA_signal 2760..2984 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 3030..3458 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3472..3801 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3868..4467 /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" polyA_signal 4641..4762 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4811..4827) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4835..4851 /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(4859..4889) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4904..4925 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5213..5798) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5969..6829) /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(6830..6934) /gene="bla" /label="AmpR promoter" ORIGIN 1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt 61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct 121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg 181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta 241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg 301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc 361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa 421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca 481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta 541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt 601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg 661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca 721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca 781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc 841 actataggga gacccaagct ggctagcgtt taaacttaag cttggtaccg agctcggatc 901 cgccaccatg ccaaaaaaga agagaaaggt tccaaaaaag aagagaaagg tcccaaaaaa 961 gaagagaaag gtaatgtcgg acacgtggag ctctatccag gcccacaaga agcagctgga 1021 ctctctgcgg gagaggctgc agcggaggcg gaagcaggac tcggggcact tggatctacg 1081 gaatccagag gcagcattgt ctccaacctt ccgtagtgac agcccagtgc ctactgcacc 1141 cacctctggt ggccctaagc ccagcacagc ttcagcagtt cctgaattag ctacagatcc 1201 tgagttagag aagaagttgc tacaccacct ctctgatctg gccttaacat tgcccactga 1261 tgctgtgtcc atctgtcttg ccatctccac gccagatgct cctgccactc aagatggggt 1321 agaaagcctc ctgcagaagt ttgcagctca ggagttgatt gaggtaaagc gaggtctcct 1381 acaagatgat gcacatccta ctcttgtaac ctatgctgac cattccaagc tctctgccat 1441 gatgggtgct gtggcagaaa agaagggccc tggggaggta gcagggactg tcacagggca 1501 gaagcggcgt gcagaacagg actcgactac agtagctgcc tttgccagtt cgttagtctc 1561 tggtctgaac tcttcagcat cggaaccagc aaaggagcca gccaagaaat caaggaaaca 1621 tgctgcctca gatgttgatc tggagataga gagccttctg aaccaacagt ccactaagga 1681 acaacagagc aagaaggtca gtcaggagat cctagagcta ttaaatacta caacagccaa 1741 ggaacaatcc attgttgaaa aatttcgctc tcgaggtcgg gcccaagtgc aagaattctg 1801 tgactatgga accaaggagg agtgcatgaa agccagtgat gctgatcgac cctgtcgcaa 1861 gctgcacttc agacgaatta tcaataaaca cactgatgag tctttaggtg actgctcttt 1921 ccttaataca tgtttccaca tggatacctg caagtatgtt cactatgaaa ttgatgcttg 1981 catggattct gaggcccctg gcagcaaaga ccacacgcca agccaggagc ttgctcttac 2041 acagagtgtc ggaggtgatt ccagtgcaga ccgactcttc ccacctcagt ggatctgttg 2101 tgatatccgc tacctggacg tcagtatctt gggcaagttt gcagttgtga tggctgaccc 2161 accctgggat attcacatgg aactgcccta tgggaccctg acagatgatg agatgcgcag 2221 gctcaacata cccgtactac aggatgatgg ctttctcttc ctctgggtca caggcagggc 2281 catggagttg gggagagaat gtctaaacct ctgggggtat gaacgggtag atgaaattat 2341 ttgggtgaag acaaatcaac tgcaacgcat cattcggaca ggccgtacag gtcactggtt 2401 gaaccatggg aaggaacact gcttggttgg tgtcaaagga aatccccaag gcttcaacca 2461 gggtctggat tgtgatgtga tcgtagctga ggttcgttcc accagtcata aaccagatga 2521 aatctatggc atgattgaaa gactatctcc tggcactcgc aagattgagt tatttggacg 2581 accacacaat gtgcaaccca actggatcac ccttggaaac caactggatg ggatccacct 2641 actagaccca gatgtggttg cacggttcaa gcaaaggtac ccagatggta tcatctctaa 2701 acctaagaat ttatagctcg agtctagagg gcccgtttaa acccgctgat cagcctcgac 2761 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct 2821 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct 2881 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg 2941 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag 3001 aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc 3061 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc 3121 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa 3181 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact 3241 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt 3301 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa 3361 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt 3421 aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag 3481 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc 3541 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa 3601 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc 3661 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat 3721 gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt 3781 ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat 3841 caagagacag gatgaggatc gtttcgcatg accgagtaca agcccacggt gcgcctcgcc 3901 acccgcgacg acgtcccccg ggccgtacgc accctcgccg ccgcgttcgc cgactacccc 3961 gccacgcgcc acaccgtcga cccggaccgc cacatcgagc gggtcaccga gctgcaagaa 4021 ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc 4081 gcggtggcgg tctggaccac gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc 4141 ggcccgcgca tggccgagtt gagcggttcc cggctggccg cgcagcaaca gatggaaggc 4201 ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc tggccaccgt cggcgtctcg 4261 cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc 4321 gagcgcgccg gggtgcccgc cttcctggag acctccgcgc cccgcaacct ccccttctac 4381 gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg 4441 tgcatgaccc gcaagcccgg tgcctgagcg ggactctggg gttcgaaatg accgaccaag 4501 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg 4561 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc 4621 tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca 4681 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt 4741 ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg 4801 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca 4861 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca 4921 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc 4981 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt 5041 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact 5101 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag 5161 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata 5221 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc 5281 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg 5341 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc 5401 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg 5461 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc 5521 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga 5581 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg 5641 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa 5701 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ttttttgttt 5761 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta 5821 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat 5881 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa 5941 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct 6001 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta 6061 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct 6121 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg 6181 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa 6241 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt 6301 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta 6361 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca 6421 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta 6481 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct 6541 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg 6601 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac 6661 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact 6721 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa 6781 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt 6841 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat 6901 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg 6961 acgtcgacgg atcgggagat ctcccgatcc cctatggt //