LOCUS V034690 7744 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V034690 VERSION V034690 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..3234 /label="Nsun2(NM_145354)" /note="Nsun2(NM_145354)" /gene="Nsun2" CDS 3241..3306 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 3344..3568 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 3614..4042 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 4056..4385 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4452..5246 /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 5420..5541 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5590..5606) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5614..5630 /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(5638..5668) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5683..5704 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5992..6577) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6748..7608) /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(7609..7713) /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 accatggggc ggcgggcgcg gggccggcgg ttccagcagc cgccgcagcc tgagggcgag 1021 gaagacgcca gcgacggcgg cagaaagcga ggccaggcgg gctgggaagg tggctatccc 1081 gagatcgtaa aggagaacaa gctcttcgag cactactatc aggaactcaa gatcgtgcca 1141 gagggagaat gggaccaatt catggagtca ctccgagaac ctctcccagc cacactgaga 1201 atcactgggt acaaaagcca tgccaaagag attctccatt gcttgaagaa caagtacttt 1261 aaggagttgg aggacctgga agtagatgga cagaaagttg aagttccaca accactaagc 1321 tggtaccctg aagaacttgc ctggcataca aacttaagtc ggaaaatctt gaggaagtcc 1381 ccgttgttgg caaagttcca tcagttcctg gtcagcgaga ctgagtctgg aaacatcagc 1441 cgccaggagg ctgtcagcat gatcccccca ctgctgctca acgtggagcc acaccataag 1501 atcttagaca tgtgtgcagc ccctggatcc aagaccacac agttaattga aatgttgcat 1561 gcagacatga gtgtgccctt tccagaggga tttgtaatcg caaatgacgt ggacaacaag 1621 cgctgctatc tgctcgtcca tcaggccaaa aggttgagca gtccctgcat catggtggta 1681 aaccatgacg catccagcat acctagactt acagtagatg tggacggaag gaaagagatt 1741 ctcttctatg atcgaatttt atgtgatgtc ccttgcagtg gcgatggcac aatgagaaaa 1801 aacattgatg tctggaagaa atggacaacc ttaaacagct tgcagctcca tggcctgcag 1861 cttcggattg caactcgagg tgctgagcag ctggcggaag gtggcaggat ggtgtattcc 1921 acgtgttcct tgaaccccgt ggaggatgaa gcagtgatcg cagctctgct agagaagagt 1981 gaaggagctc ttgagcttgc tgatgtgtct gctgagttgc caggactgaa gtggatgcct 2041 ggagtctcac agtggaaggt catgactaga gacgggcagt ggtttgcaga ctggcatgag 2101 gttccccagg gcaggcatac acaaatccga cctaccatgt tcccaccaac ggacctggag 2161 aagctacagg caatgcatct agagcgatgc cttcgaatcc tgccccatca tcagaatact 2221 ggagggttct ttgtggcagt attggtcaag aaagcaccaa tgccgtggaa caaacgtcag 2281 cccaaggtcc agaataaatc tgcagaagcc agagaaccca gggtatccag ccatgtggct 2341 gccacagagg gaaatcccag tgaccagtct gagctggaaa gtcagatgat aactggagct 2401 ggtgactcag aaacagctca caacactgag aacacagaga gcaatgagaa gaaagatggc 2461 gtgtgtggcc ctcctccatc aaagaaaatg aagttgtttg gatttaaaga agatccattt 2521 gtattcattc ctgaagatga tcctttattt ccacctattg agaagtttta tgccttggat 2581 ccttcattcc cgaggatgaa tctgttaacc cgaaccacag aaggaaagaa gcggcagctt 2641 tatatggtct ccaaggagct gaggaatgta ctgctgaaca acagcgagaa gatgaaggtc 2701 attaacactg ggataaaagt ctggtgtcga aataacagtg gtgaagaatt cgattgtgca 2761 ttccgtttgg cacaggaggg aatatataca ttgtatccat ttatcaattc aagaatcatc 2821 actgtatcaa tggaagacgt gaagacactg ttgacccagg agaacccatt ctttagaaaa 2881 ctgagcagtg aggcctacag tcaagtcaag gaccttgcaa agggaagtgt tgtgctgaag 2941 tatgagccag attctgcgaa tccagacacc ctccagtgcc ccatcgtgct gtgtgggtgg 3001 cggggaaagg cctctattcg aacttttgtg cccaaaaatg agcggcttca ttatctcagg 3061 atgatgggtc tggaggtact gggagaaaag aagaaagagg gagtcattct taccaatgag 3121 aatgctgcca gcccagagca gcctggagat gaggatgcca agcagacagc acaagacccc 3181 tgcgtcccag actccgtccc tggctgtgat gcagctgcag ctgagccatc ccggctcgag 3241 gactacaaag accatgacgg tgattataaa gatcatgaca tcgactacaa ggatgacgat 3301 gacaagtagt gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc 3361 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc 3421 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct 3481 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg 3541 catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct 3601 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg 3661 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct 3721 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta 3781 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt 3841 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg 3901 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat 3961 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt 4021 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt 4081 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca 4141 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt 4201 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt 4261 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg 4321 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt 4381 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag 4441 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg 4501 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt 4561 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc 4621 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt 4681 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag 4741 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg 4801 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag 4861 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg 4921 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc 4981 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca 5041 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc 5101 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg 5161 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct 5221 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc 5281 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg 5341 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct 5401 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa 5461 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc 5521 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc 5581 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa 5641 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac 5701 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca 5761 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc 5821 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc 5881 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc 5941 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag 6001 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc 6061 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt 6121 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct 6181 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg 6241 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct 6301 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat 6361 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg 6421 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa 6481 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtt tttttgtttg 6541 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac 6601 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc 6661 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag 6721 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc 6781 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac 6841 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc 6901 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg 6961 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag 7021 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc 7081 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac 7141 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag 7201 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac 7261 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg 7321 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc 7381 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact 7441 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg 7501 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa 7561 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt 7621 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg 7681 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga 7741 cgtc //