LOCUS       sequence-227046-        6999 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cmv d0 dPspCas13b-GGS-Sun no-fusion construct.
ACCESSION   .
VERSION     .
KEYWORDS    .
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 Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1021/jacs.8b05012"; journalName: "J Am Chem Soc"; date: 
            "2018-09-26- 26"; volume: "140"; issue: "38"; pages: "11974-11981"
COMMENT     Sequence Label: Cmv d0 dPspCas13b-GGS-Sun
FEATURES             Location/Qualifiers
     source          1..6999
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    191..474
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
     enhancer        665..1044
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1045..1248
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1199..1219
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          1384..1516
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1499..1518
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             4597..4653
                     /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             4669..4725
                     /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             4741..4797
                     /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             4804..4827
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(4945..4963)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    5000..5121
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5011..5030)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5065..5084
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             5520..6314
                     /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(5574..5593)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6184..6203
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(join(6486..6999,1..75))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6566..6585)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
       61 aaaaaaggat ttcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
      121 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
      181 cttttatagt ccggaaatac aggaacgcac gctggatggc ccttcgctgg gatggtgaaa
      241 ccatgaaaaa tggcagcttc agtggattaa gtgggggtaa tgtggcctgt accctctggt
      301 tgcataggta ttcatacggt taaaatttat caggcgcgat tgcggcagtt tttcgggtgg
      361 tttgttgcca tttttacctg tctgctgccg tgatcgcgct gaacgcgttt tagcggtgcg
      421 tacaattaag ggattatggt aaatccactt actgtctgcc ctcgtagcca tcgagataaa
      481 ccgcagtact ccggccacga tgcgtccggc gtagaggatc gagatcttca atattggcca
      541 ttagccatat tattcattgg ttatatagca taaatcaata ttggctattg gccattgcat
      601 acgttgtatc tatatcataa tatgtacatt tatattggct catgtccaat atgaccgcca
      661 tgttggcatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
      721 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      781 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      841 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      901 catcaagtgt atcatatgcc aagtccgccc cctattgacg tcaatgacgg taaatggccc
      961 gcctggcatt atgcccagta catgacctta cgggactttc ctacttggca gtacatctac
     1021 gtattagtca tcgctattac catggtgatg cggttttggc agtacaccaa tgggcgtgga
     1081 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
     1141 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta ataaccccgc cccgttgacg
     1201 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctgg tttagtgaac
     1261 cgtcagatca ctagaagctt tattgcggta gtttatcaca gttaaattgc taacgcagtc
     1321 agtgcttctg acacaacagt ctcgaactta agctgcagaa gttggtcgtg aggcactggg
     1381 caggtaagta tcaaggttac aagacaggtt taaggagacc aatagaaact gggcttgtcg
     1441 agacagagaa gactcttgcg tttctgatag gcacctattg gtcttactga catccacttt
     1501 gcctttctct ccacaggtgt ccactcccag ttcaattaca gctcttaagg cagatcacta
     1561 gaagcttacg ctaccaccat gaacatcccc gctctggtgg aaaaccagaa gaagtacttt
     1621 ggcacctaca gcgtgatggc catgctgaac gctcagaccg tgctggacca catccagaag
     1681 gtggccgata ttgagggcga gcagaacgag aacaacgaga atctgtggtt tcaccccgtg
     1741 atgagccacc tgtacaacgc caagaacggc tacgacaagc agcccgagaa aaccatgttc
     1801 atcatcgagc ggctgcagag ctacttccca ttcctgaaga tcatggccga gaaccagaga
     1861 gagtacagca acggcaagta caagcagaac cgcgtggaag tgaacagcaa cgacatcttc
     1921 gaggtgctga agcgcgcctt cggcgtgctg aagatgtaca gggacctgac caacgcatac
     1981 aagacctacg aggaaaagct gaacgacggc tgcgagttcc tgaccagcac agagcaacct
     2041 ctgagcggca tgatcaacaa ctactacaca gtggccctgc ggaacatgaa cgagagatac
     2101 ggctacaaga cagaggacct ggccttcatc caggacaagc ggttcaagtt cgtgaaggac
     2161 gcctacggca agaaaaagtc ccaagtgaat accggattct tcctgagcct gcaggactac
     2221 aacggcgaca cacagaagaa gctgcacctg agcggagtgg gaatcgccct gctgatctgc
     2281 ctgttcctgg acaagcagta catcaacatc tttctgagca ggctgcccat cttctccagc
     2341 tacaatgccc agagcgagga acggcggatc atcatcagat ccttcggcat caacagcatc
     2401 aagctgccca aggaccggat ccacagcgag aagtccaaca agagcgtggc catggatatg
     2461 ctcaacgaag tgaagcggtg ccccgacgag ctgttcacaa cactgtctgc cgagaagcag
     2521 tcccggttca gaatcatcag cgacgaccac aatgaagtgc tgatgaagcg gagcagcgac
     2581 agattcgtgc ctctgctgct gcagtatatc gattacggca agctgttcga ccacatcagg
     2641 ttccacgtga acatgggcaa gctgagatac ctgctgaagg ccgacaagac ctgcatcgac
     2701 ggccagacca gagtcagagt gatcgagcag cccctgaacg gcttcggcag actggaagag
     2761 gccgagacaa tgcggaagca agagaacggc accttcggca acagcggcat ccggatcaga
     2821 gacttcgaga acatgaagcg ggacgacgcc aatcctgcca actatcccta catcgtggac
     2881 acctacacac actacatcct ggaaaacaac aaggtcgaga tgtttatcaa cgacaaagag
     2941 gacagcgccc cactgctgcc cgtgatcgag gatgatagat acgtggtcaa gacaatcccc
     3001 agctgccgga tgagcaccct ggaaattcca gccatggcct tccacatgtt tctgttcggc
     3061 agcaagaaaa ccgagaagct gatcgtggac gtgcacaacc ggtacaagag actgttccag
     3121 gccatgcaga aagaagaagt gaccgccgag aatatcgcca gcttcggaat cgccgagagc
     3181 gacctgcctc agaagatcct ggatctgatc agcggcaatg cccacggcaa ggatgtggac
     3241 gccttcatca gactgaccgt ggacgacatg ctgaccgaca ccgagcggag aatcaagaga
     3301 ttcaaggacg accggaagtc cattcggagc gccgacaaca agatgggaaa gagaggcttc
     3361 aagcagatct ccacaggcaa gctggccgac ttcctggcca aggacatcgt gctgtttcag
     3421 cccagcgtga acgatggcga gaacaagatc accggcctga actaccggat catgcagagc
     3481 gccattgccg tgtacgatag cggcgacgat tacgaggcca agcagcagtt caagctgatg
     3541 ttcgagaagg cccggctgat cggcaagggc acaacagagc ctcatccatt tctgtacaag
     3601 gtgttcgccc gcagcatccc cgccaatgcc gtcgagttct acgagcgcta cctgatcgag
     3661 cggaagttct acctgaccgg cctgtccaac gagatcaaga aaggcaacag agtggatgtg
     3721 cccttcatcc ggcgggacca gaacaagtgg aaaacacccg ccatgaagac cctgggcaga
     3781 atctacagcg aggatctgcc cgtggaactg cccagacaga tgttcgacaa tgagatcaag
     3841 tcccacctga agtccctgcc acagatggaa ggcatcgact tcaacaatgc caacgtgacc
     3901 tatctgatcg ccgagtacat gaagagagtg ctggacgacg acttccagac cttctaccag
     3961 tggaaccgca actaccggta catggacatg cttaagggcg agtacgacag aaagggctcc
     4021 ctgcagcact gcttcaccag cgtggaagag agagaaggcc tctggaaaga gcgggcctcc
     4081 agaacagagc ggtacagaaa gcaggccagc aacaagatcc gcagcaaccg gcagatgaga
     4141 aacgccagca gcgaagagat cgagacaatc ctggataagc ggctgagcaa cagccggaac
     4201 gagtaccaga aaagcgagaa agtgatccgg cgctacagag tgcaggatgc cctgctgttt
     4261 ctgctggcca aaaagaccct gaccgaactg gccgatttcg acggcgagag gttcaaactg
     4321 aaagaaatca tgcccgacgc cgagaaggga atcctgagcg agatcatgcc catgagcttc
     4381 accttcgaga aaggcggcaa gaagtacacc atcaccagcg agggcatgaa gctgaagaac
     4441 tacggcgact tctttgtgct ggctagcgac aagaggatcg gcaacctgct ggaactcgtg
     4501 ggcagcgaca tcgtgtccaa agaggatggc ggtagtggag gaagcggggg gagcggaggg
     4561 agtggtggat ccggcggctc cgggtccgga tcgggggaag aacttttgag caagaattat
     4621 catcttgaga acgaagtggc tcgtcttaag aaaggttctg gcagtggaga agaactgctt
     4681 tcaaagaatt accacctgga aaatgaggta gctagactga aaaaggggag cggaagtggg
     4741 gaggagttgc tgagcaaaaa ttatcatttg gagaacgaag tagcacgact aaagaaagga
     4801 tccgattaca aggacgacga tgacaagtag tgtttaaacg aattcgggct cggtacacgg
     4861 ggatcctcta gagtcgacct gcaggcatgc aagctgatcc ggctgctaac aaagcccgaa
     4921 aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc cttggggcgg
     4981 ccgcttcgag cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg
     5041 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt
     5101 ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag
     5161 ggggagatgt gggaggtttt tttaagcaag taaaacctct acaaatgtgg taaaatcgaa
     5221 ttctaatgga tcctctttgc gcttgcgttt tcccttgtcc agatagccca gtagctgaca
     5281 ttcatccggg gtcagcaccg tttctgcgga ctggctttct acgtgttccg cttcctttag
     5341 cagcccttgc gccctgagtg cttgcggcag cgtgagcttc aaaagaattg ccagctgggg
     5401 cgccctctgg taaggttggg aagccctgca aagtaaactg gatggctttc ttgccgccaa
     5461 ggatctgatg gcgcagggga tcaagatctg atcaagagac aggatgacgg tcgtttcgca
     5521 tgcttgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     5581 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     5641 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     5701 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     5761 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     5821 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     5881 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     5941 tcgagcgagc acgcactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     6001 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgt atgccggatg
     6061 gtgaggatct cgtcgtgact catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     6121 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     6181 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     6241 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     6301 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaaccg
     6361 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     6421 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     6481 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     6541 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     6601 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     6661 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     6721 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     6781 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     6841 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     6901 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca
     6961 gttaccttcg gaaaaagagt tggtagctct tgatccggc
//
