LOCUS V043133 7845 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V043133 VERSION V043133 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 927..2585 /label="Mettl16(NM_026197)" /note="Mettl16(NM_026197)" /gene="Mettl16" CDS 2592..2657 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" CDS 2688..3407 /codon_start=1 /note="mammalian codon-optimized" /product="the original enhanced GFP (Yang et al., 1996)" /transl_table=1 /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL TLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV LLEFVTAAGITLGMDELYK*" /label="EGFP" polyA_signal 3445..3669 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 3715..4143 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 4157..4486 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4553..5347 /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 5521..5642 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5691..5707) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5715..5731 /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(5739..5769) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5784..5805 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6093..6678) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6849..7709) /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(7710..7814) /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 gtttaaagcc actgggatcc gccaccatgg ctctaagcaa atcaatgcat gcaagaaata 961 gatacaagga caaaccacct gacttcgcat atctggcatc taaatatcca gattttaagc 1021 agcatattca gataaatctg aatgggagag taagtcttaa ttttaaagac cccgaagcag 1081 tcagagctct gacttgtact ctcctgagag aggactttgg actttctatt gatattcctt 1141 tggagagact aattcccaca gtccccttga gacttaacta tattcactgg gtagaagatt 1201 tgattggtca ccaggattcg gacaaaacta ctctccgaag aggaattgac ataggtactg 1261 gggcatcctg catctatccc ttacttggag caaccttaaa tggctggtat ttcctggcaa 1321 cagaagtgga tgatatgtgt ttcaattatg ccaagaaaaa tgtagagcag aataacctat 1381 ctgatctcat taaagtggtg aaagtgccgc agaagacact cctgatggat gcgcttaagg 1441 aagagtctga gatagtctat gacttctgta tgtgcaaccc tccctttttt gccaaccaat 1501 tggaagccaa gggtgtaaat tcccggaatt ctcgaagacc tccacctagt tctgtaaaca 1561 ccgggggcat cacagagatt atggcagaag gaggtgagct agaatttgtc aaaaggatca 1621 tccatgacag tttacagctc aaaaaaaggt tacggtggta tagctgtatg ctgggaaaga 1681 aatgcagtct ggctccactg aaggaagagc ttagaatcca aggggttcct aaagtcacct 1741 tcacagagtt ttgtcagggt cggacaatga gatgggcctt agcttggagt ttttatgatg 1801 atgtcacagt accgtcacct ccaagtaaac gaagaaaatt agaaaagcca aggaaaccca 1861 ttacttttgt agtgttggag tctgtgatga aagaattgtc tctcaaagct tcatctttgg 1921 gctccgagac agcggaaggc atagtggttg taacaacatg gatagaaaag atcctgactg 1981 atctgaaggt gcaacataag cgaattccct gtggaagaga agaagttagc cttttcctga 2041 cagccataga aaactcttgg attcatttaa ggcggaagag aagagagcga gtgaggcagc 2101 tgcgggaggt gcctcgggct cctgaggacg tcatcttagc cttggaagag agaaagtcca 2161 ctcccaaaga gctgagtagt ggccaagacg tggcccacag cccccaagag agtgccctgt 2221 gtgggcttga tgtgccgggc ggcgaggctg ctgctgatgg aggccattgc ctcagccaga 2281 agttactgtg ccaggaggag actcccgagg ccacggagga tgaaagggat gaggaaaggg 2341 gagggatgga ggccatggaa agttgtaaag gctccagcaa tggagcccag gacggagagg 2401 cttctgagaa gggggatcgc ctggatgggg cggccggacg ctacctgttt aagtgtttag 2461 tcaacattaa gaaggaagct ggcgatgccg tggttgaaat gcattgggtt gagggccaga 2521 atagggattt gatgaaccag ctttgcactt acgtacgtaa ccaaattttg aggcttgttg 2581 ctagtctcga ggactacaaa gaccatgacg gtgattataa agatcatgac atcgactaca 2641 aggatgacga tgacaagggc ggtggaggaa gcggcggagg tggatctatg gtgagcaagg 2701 gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg 2761 gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc 2821 tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc 2881 tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct 2941 tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg 3001 gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg 3061 agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca 3121 actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga 3181 acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc 3241 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc 3301 agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg 3361 tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtagtct agagggcccg 3421 tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc 3481 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa 3541 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg 3601 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg 3661 gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc 3721 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac 3781 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg 3841 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt 3901 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc 3961 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct 4021 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga 4081 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga 4141 attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg 4201 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg 4261 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc 4321 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca 4381 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt 4441 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc 4501 ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga 4561 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga 4621 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg 4681 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga 4741 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt 4801 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct 4861 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct 4921 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg 4981 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca 5041 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga 5101 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt 5161 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt 5221 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct 5281 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt 5341 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca 5401 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg 5461 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc 5521 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca 5581 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct 5641 tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg 5701 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata 5761 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca 5821 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc 5881 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg 5941 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta 6001 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc 6061 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag 6121 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac 6181 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc 6241 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt 6301 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc 6361 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga 6421 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta 6481 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta 6541 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga 6601 tccggcaaac aaaccaccgc tggtagcggt ttttttgttt gcaagcagca gattacgcgc 6661 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg 6721 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag 6781 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg 6841 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt 6901 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca 6961 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca 7021 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc 7081 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt 7141 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg 7201 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc 7261 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg 7321 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga 7381 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga 7441 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta 7501 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg 7561 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact 7621 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata 7681 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt 7741 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa 7801 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtc //