LOCUS V033651 6841 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V033651 VERSION V033651 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers enhancer 235..614 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 615..818 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" promoter 863..881 /note="promoter for bacteriophage T7 RNA polymerase" /label="T7 promoter" CDS 964..2331 /label="Mettl14(NM_201638)" /note="Mettl14(NM_201638)" /gene="Mettl14" CDS 2338..2403 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 2441..2665 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2711..3139 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3153..3482 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3549..4343 /codon_start=1 /gene="aph(3')-II (or nptII)" /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin®)" /product="aminoglycoside phosphotransferase from Tn5" /transl_table=1 /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*" /label="NeoR/KanR" polyA_signal 4517..4638 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4687..4703) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4711..4727 /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(4735..4765) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4780..4801 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5089..5674) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5845..6705) /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(6706..6810) /gene="bla" /label="AmpR promoter" ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg 61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt 241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata 301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc 361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc 421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt 481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt 541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca 601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg 661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc 721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg 781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca 841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc 901 gtttaaactt aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattcgcc 961 accatggata gccgcctgca ggagatccgg gagcggcaga agttacggcg gcagctccta 1021 gctcagcagt tgggagctga gagtgcggat agcattggtg ctgtgttaaa tagcaaagat 1081 gaacagaggg agattgctga aaccagagaa acctgcaggg cttcctatga tacatctgct 1141 ccaaactcaa aacggaagtg tctggatgaa ggagagactg atgaagacaa agtagaagaa 1201 tataaggatg aactggaaat gcagcaggag gaagagaatt tgccatatga agaagagatt 1261 tacaaagatt ccagtacctt tcttaaggga acgcagagct taaatcccca taatgattac 1321 tgccaacatt ttgtagacac tggacacaga cctcagaatt tcatcaggga tgtaggttta 1381 gctgacagat ttgaagaata ccctaaactt agggaactca tcagactaaa ggatgagtta 1441 atagctaagt caaacactcc tcccatgtac ttacaagcag acatagaagc ctttgacatc 1501 agagaattga cacccaaatt tgatgtgatt ctcctggagc cccctctgga agaatactat 1561 agagagactg gcatcactgc gaatgagaaa tgctggacct gggatgatat tatgaagtta 1621 gaaatcgatg agattgcagc acctcggtca tttatatttc tctggtgcgg ttctggggaa 1681 ggattggacc ttgggagagt atgcttgcga aagtggggtt acagaagatg tgaagatatt 1741 tgttggatta aaaccaataa aaacaatcct ggaaagacaa agactctaga tccaaaggca 1801 gttttccaga gaacaaagga gcattgcctg atggggatca aaggaaccgt gaagcgaagc 1861 acagacgggg acttcattca tgctaatgtt gacattgact taattatcac agaagaacct 1921 gagattggca atatagaaaa accagtagaa atttttcata taatagaaca tttttgtctt 1981 ggtagaagac gccttcatct ctttgggaga gatagcacta tcaggccagg ctggctcaca 2041 gttggaccaa cgcttacaaa cagtaactac aatgcagaaa catatgcatc gtatttcagt 2101 gcccccaact catacttgac cggatgtaca gaggaaatcg agaggcttcg accgaagtca 2161 cctcctccca agtccaagtc tgaccgtggg ggtggagctc ccagaggtgg aggaaggggg 2221 ggaacatctg ctggccgtgg tcgggaaaga aaccgatcca atttccgagg agagagaggt 2281 ggctttaggg ggggccgtgg aggcacgcac agaggcggct ttactcctcg gctcgaggac 2341 tacaaagacc atgacggtga ttataaagat catgacatcg actacaagga tgacgatgac 2401 aagtagtgag ggcccgttta aacccgctga tcagcctcga ctgtgccttc tagttgccag 2461 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact 2521 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt 2581 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat 2641 gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg 2701 gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc 2761 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc 2821 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg 2881 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca 2941 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc 3001 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct 3061 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa 3121 caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc 3181 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt 3241 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt 3301 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg 3361 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct 3421 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca 3481 aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat 3541 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga 3601 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc 3661 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga 3721 atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg 3781 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc 3841 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg 3901 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga 3961 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc 4021 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca 4081 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg 4141 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct 4201 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg 4261 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc 4321 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac 4381 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt 4441 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga 4501 gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag 4561 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa 4621 actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta 4681 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat 4741 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt 4801 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta 4861 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc 4921 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa 4981 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa 5041 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct 5101 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac 5161 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc 5221 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc 5281 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg 5341 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga 5401 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag 5461 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta 5521 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag 5581 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggttttt ttgtttgcaa 5641 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg 5701 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa 5761 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat 5821 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc 5881 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat 5941 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc 6001 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc 6061 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag 6121 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg 6181 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg 6241 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag 6301 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt 6361 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga 6421 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc 6481 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc 6541 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc 6601 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc 6661 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca 6721 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat 6781 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt 6841 c //