LOCUS Exported 6999 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Cmv d0 dPspCas13b-GGS-Sun no-fusion construct. ACCESSION . VERSION . KEYWORDS Cmv d0 dPspCas13b-GGS-Sun SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6999) AUTHORS Rauch S, He C, Dickinson BC TITLE Targeted m(6)A Reader Proteins To Study Epitranscriptomic Regulation of Single RNAs. JOURNAL J Am Chem Soc. 2018 Sep 26;140(38):11974-11981. doi: 10.1021/jacs.8b05012. Epub 2018 Sep 18. PUBMED 30183280 REFERENCE 2 (bases 1 to 6999) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6999 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 69..448 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 449..652 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 603..623 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" intron 788..920 /label=chimeric intron /note="chimera between introns from human beta-globin and immunoglobulin heavy chain genes" primer_bind 903..922 /label=pMT2-F /note="Synthetic intron, forward primer" CDS 4001..4057 /codon_start=1 /product="GCN4 peptide optimized to improve solubility while preserving binding to an scFv-GFP fusion protein (Tanenbaum et al., 2014)" /label=GCN4_v4 /translation="EELLSKNYHLENEVARLKK" CDS 4073..4129 /codon_start=1 /product="GCN4 peptide optimized to improve solubility while preserving binding to an scFv-GFP fusion protein (Tanenbaum et al., 2014)" /label=GCN4_v4 /translation="EELLSKNYHLENEVARLKK" CDS 4145..4201 /codon_start=1 /product="GCN4 peptide optimized to improve solubility while preserving binding to an scFv-GFP fusion protein (Tanenbaum et al., 2014)" /label=GCN4_v4 /translation="EELLSKNYHLENEVARLKK" CDS 4208..4231 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" primer_bind complement(4349..4367) /label=T7 Term /note="T7 terminator, reverse primer" polyA_signal 4404..4525 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4415..4434) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 4469..4488 /label=SV40pA-R /note="SV40 polyA, reverse primer" CDS 4924..5718 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MLEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(4978..4997) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 5588..5607 /label=Neo-F /note="Neomycin resistance gene, forward primer" rep_origin complement(5890..6478) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5970..5989) /label=pBR322ori-F /note="pBR322 origin, forward primer" misc_feature 6594..6877 /label=cer region /note="ColE1-derived recombination site that helps to maintain plasmids as monomers" ORIGIN 1 gcatacgttg tatctatatc ataatatgta catttatatt ggctcatgtc caatatgacc 61 gccatgttgg cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt 121 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg 181 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc 241 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc 301 agtacatcaa gtgtatcata tgccaagtcc gccccctatt gacgtcaatg acggtaaatg 361 gcccgcctgg cattatgccc agtacatgac cttacgggac tttcctaatt ggcagtacat 421 ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca ccaatgggcg 481 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag 541 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaataacc ccgccccgtt 601 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctggtttagt 661 gaaccgtcag atcactagaa gctttattgc ggtagtttat cacagttaaa ttgctaacgc 721 agtcagtgct tctgacacaa cagtctcgaa cttaagctgc agaagttggt cgtgaggcac 781 tgggcaggta agtatcaagg ttacaagaca ggtttaagga gaccaataga aactgggctt 841 gtcgagacag agaagactct tgcgtttctg ataggcacct attggtctta ctgacatcca 901 ctttgccttt ctctccacag gtgtccactc ccagttcaat tacagctctt aaggcagatc 961 actagaagct tacgctacca ccatgaacat ccccgctctg gtggaaaacc agaagaagta 1021 ctttggcacc tacagcgtga tggccatgct gaacgctcag accgtgctgg accacatcca 1081 gaaggtggcc gatattgagg gcgagcagaa cgagaacaac gagaatctgt ggtttcaccc 1141 cgtgatgagc cacctgtaca acgccaagaa cggctacgac aagcagcccg agaaaaccat 1201 gttcatcatc gagcggctgc agagctactt cccattcctg aagatcatgg ccgagaacca 1261 gagagagtac agcaacggca agtacaagca gaaccgcgtg gaagtgaaca gcaacgacat 1321 cttcgaggtg ctgaagcgcg ccttcggcgt gctgaagatg tacagggacc tgaccaacgc 1381 atacaagacc tacgaggaaa agctgaacga cggctgcgag ttcctgacca gcacagagca 1441 acctctgagc ggcatgatca acaactacta cacagtggcc ctgcggaaca tgaacgagag 1501 atacggctac aagacagagg acctggcctt catccaggac aagcggttca agttcgtgaa 1561 ggacgcctac ggcaagaaaa agtcccaagt gaataccgga ttcttcctga gcctgcagga 1621 ctacaacggc gacacacaga agaagctgca cctgagcgga gtgggaatcg ccctgctgat 1681 ctgcctgttc ctggacaagc agtacatcaa catctttctg agcaggctgc ccatcttctc 1741 cagctacaat gcccagagcg aggaacggcg gatcatcatc agatccttcg gtatcaacag 1801 catcaagctg cccaaggacc ggatccacag cgagaagtcc aacaagagcg tggccatgga 1861 tatgctcaac gaagtgaagc ggtgccccga cgagctgttc acaacactgt ctgccgagaa 1921 gcagtcccgg ttcagaatca tcagcgacga ccacaatgaa gtgctgatga agcggagcag 1981 cgacagattc gtgcctctgc tgctgcagta tatcgattac ggcaagctgt tcgaccacat 2041 caggttccac gtgaacatgg gcaagctgag atacctgctg aaggccgaca agacctgcat 2101 cgacggccag accagagtca gagtgatcga gcagcccctg aacggcttcg gcagactgga 2161 agaggccgag acaatgcgga agcaagagaa cggcaccttc ggcaacagcg gcatccggat 2221 cagagacttc gagaacatga agcgggacga cgccaatcct gccaactatc cctacatcgt 2281 ggacacctac acacactaca tcctggaaaa caacaaggtc gagatgttta tcaacgacaa 2341 agaggacagc gccccactgc tgcccgtgat cgaggatgat agatacgtgg tcaagacaat 2401 ccccagctgc cggatgagca ccctggaaat tccagccatg gccttccaca tgtttctgtt 2461 cggcagcaag aaaaccgaga agctgatcgt ggacgtgcac aaccggtaca agagactgtt 2521 ccaggccatg cagaaagaag aagtgaccgc cgagaatatc gccagcttcg gaatcgccga 2581 gagcgacctg cctcagaaga tcctggatct gatcagcggc aatgcccacg gcaaggatgt 2641 ggacgccttc atcagactga ccgtggacga catgctgacc gacaccgagc ggagaatcaa 2701 gagattcaag gacgaccgga agtccattcg gagcgccgac aacaagatgg gaaagagagg 2761 cttcaagcag atctccacag gcaagctggc cgacttcctg gccaaggaca tcgtgctgtt 2821 tcagcccagc gtgaacgatg gcgagaacaa gatcaccggc ctgaactacc ggatcatgca 2881 gagcgccatt gccgtgtacg atagcggcga cgattacgag gccaagcagc agttcaagct 2941 gatgttcgag aaggcccggc tgatcggcaa gggcacaaca gagcctcatc catttctgta 3001 caaggtgttc gcccgcagca tccccgccaa tgccgtcgag ttctacgagc gctacctgat 3061 cgagcggaag ttctacctga ccggcctgtc caacgagatc aagaaaggca acagagtgga 3121 tgtgcccttc atccggcggg accagaacaa gtggaaaaca cccgccatga agaccctggg 3181 cagaatctac agcgaggatc tgcccgtgga actgcccaga cagatgttcg acaatgagat 3241 caagtcccac ctgaagtccc tgccacagat ggaaggcatc gacttcaaca atgccaacgt 3301 gacctatctg atcgccgagt acatgaagag agtgctggac gacgacttcc agaccttcta 3361 ccagtggaac cgcaactacc ggtacatgga catgcttaag ggcgagtacg acagaaaggg 3421 ctccctgcag cactgcttca ccagcgtgga agagagagaa ggcctctgga aagagcgggc 3481 ctccagaaca gagcggtaca gaaagcaggc cagcaacaag atccgcagca accggcagat 3541 gagaaacgcc agcagcgaag agatcgagac aatcctggat aagcggctga gcaacagccg 3601 gaacgagtac cagaaaagcg agaaagtgat ccggcgctac agagtgcagg atgccctgct 3661 gtttctgctg gccaaaaaga ccctgaccga actggccgat ttcgacggcg agaggttcaa 3721 actgaaagaa atcatgcccg acgccgagaa gggaatcctg agcgagatca tgcccatgag 3781 cttcaccttc gagaaaggcg gcaagaagta caccatcacc agcgagggca tgaagctgaa 3841 gaactacggc gacttctttg tgctggctag cgacaagagg atcggcaacc tgctggaact 3901 cgtgggcagc gacatcgtgt ccaaagagga tggcggtagt ggaggaagcg gggggagcgg 3961 agggagtggt ggatccggcg gctccgggtc cggatcgggg gaagaacttt tgagcaagaa 4021 ttatcatctt gagaacgaag tggctcgtct taagaaaggt tctggcagtg gagaagaact 4081 gctttcaaag aattaccacc tggaaaatga ggtagctaga ctgaaaaagg ggagcggaag 4141 tggggaggag ttgctgagca aaaattatca tttggagaac gaagtagcac gactaaagaa 4201 aggatccgat tacaaggacg acgatgacaa gtagtgttta aacgaattcg ggctcggtac 4261 acggggatcc tctagagtcg acctgcaggc atgcaagctg atccggctgc taacaaagcc 4321 cgaaaggaag ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg 4381 gcggccgctt cgagcagaca tgataagata cattgatgag tttggacaaa ccacaactag 4441 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac 4501 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt 4561 tcagggggag atgtgggagg tttttttaag caagtaaaac ctctacaaat gtggtaaaat 4621 cgaattctaa tggatcctct ttgcgcttgc gttttccctt gtccagatag cccagtagct 4681 gacattcatc cggggtcagc accgtttctg cggactggct ttctacgtgt tccgcttcct 4741 ttagcagccc ttgcgccctg agtgcttgcg gcagcgtgag cttcaaaaga attgccagct 4801 ggggcgccct ctggtaaggt tgggaagccc tgcaaagtaa actggatggc tttcttgccg 4861 ccaaggatct gatggcgcag gggatcaaga tctgatcaag agacaggatg acggtcgttt 4921 cgcatgcttg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta 4981 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg 5041 tcagcgcagg ggcgcccggt tctttttgtc aagacagacc tgtccggtgc cctgaatgaa 5101 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct 5161 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg 5221 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca 5281 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat 5341 cgcatcgagc gagcacgcac tcggatggaa gccggtcttg tcgatcagga tgatctggac 5401 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgtatgccg 5461 gatggtgagg atctcgtcgt gactcatggc gatgcctgct tgccgaatat catggtggaa 5521 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag 5581 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc 5641 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt 5701 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca 5761 accggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc 5821 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt 5881 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag 5941 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc 6001 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc 6061 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt 6121 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt 6181 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc 6241 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa 6301 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa 6361 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg 6421 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatttcaaga 6481 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg 6541 gattttggtc atgagattat caaaaaggat cttcacctag atccttttat agtccggaaa 6601 tacaggaacg cacgctggat ggcccttcgc tgggatggtg aaaccatgaa aaatggcagc 6661 ttcagtggat taagtggggg taatgtggcc tgtaccctct ggttgcatag gtattcatac 6721 ggttaaaatt tatcaggcgc gattgcggca gtttttcggg tggtttgttg ccatttttac 6781 ctgtctgctg ccgtgatcgc gctgaacgcg ttttagcggt gcgtacaatt aagggattat 6841 ggtaaatcca cttactgtct gccctcgtag ccatcgagat aaaccgcagt actccggcca 6901 cgatgcgtcc ggcgtagagg atcgagatct tcaatattgg ccattagcca tattattcat 6961 tggttatata gcataaatca atattggcta ttggccatt //