LOCUS       sequence-231774-        7053 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cmv d0 dPspCas13b-GGS-NYTHDF2(100-200) (YTDHF2 100-200 subdomain).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7053)
  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 7053)
  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-NYTHDF2(100-200)
FEATURES             Location/Qualifiers
     source          1..7053
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        65..444
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        445..648
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     599..619
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          784..916
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     899..918
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             4258..4281
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(4399..4417)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    4454..4575
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4465..4484)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4519..4538
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             4974..5768
                     /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(5028..5047)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5638..5657
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(5940..6528)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6020..6039)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    6644..6927
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
ORIGIN
        1 acgttgtatc tatatcataa tatgtacatt tatattggct catgtccaat atgaccgcca
       61 tgttggcatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
      121 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      181 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      241 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      301 catcaagtgt atcatatgcc aagtccgccc cctattgacg tcaatgacgg taaatggccc
      361 gcctggcatt atgcccagta catgacctta cgggactttc ctacttggca gtacatctac
      421 gtattagtca tcgctattac catggtgatg cggttttggc agtacaccaa tgggcgtgga
      481 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
      541 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta ataaccccgc cccgttgacg
      601 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctgg tttagtgaac
      661 cgtcagatca ctagaagctt tattgcggta gtttatcaca gttaaattgc taacgcagtc
      721 agtgcttctg acacaacagt ctcgaactta agctgcagaa gttggtcgtg aggcactggg
      781 caggtaagta tcaaggttac aagacaggtt taaggagacc aatagaaact gggcttgtcg
      841 agacagagaa gactcttgcg tttctgatag gcacctattg gtcttactga catccacttt
      901 gcctttctct ccacaggtgt ccactcccag ttcaattaca gctcttaagg cagatcacta
      961 gaagcttacg ctaccaccat gaacatcccc gctctggtgg aaaaccagaa gaagtacttt
     1021 ggcacctaca gcgtgatggc catgctgaac gctcagaccg tgctggacca catccagaag
     1081 gtggccgata ttgagggcga gcagaacgag aacaacgaga atctgtggtt tcaccccgtg
     1141 atgagccacc tgtacaacgc caagaacggc tacgacaagc agcccgagaa aaccatgttc
     1201 atcatcgagc ggctgcagag ctacttccca ttcctgaaga tcatggccga gaaccagaga
     1261 gagtacagca acggcaagta caagcagaac cgcgtggaag tgaacagcaa cgacatcttc
     1321 gaggtgctga agcgcgcctt cggcgtgctg aagatgtaca gggacctgac caacgcatac
     1381 aagacctacg aggaaaagct gaacgacggc tgcgagttcc tgaccagcac agagcaacct
     1441 ctgagcggca tgatcaacaa ctactacaca gtggccctgc ggaacatgaa cgagagatac
     1501 ggctacaaga cagaggacct ggccttcatc caggacaagc ggttcaagtt cgtgaaggac
     1561 gcctacggca agaaaaagtc ccaagtgaat accggattct tcctgagcct gcaggactac
     1621 aacggcgaca cacagaagaa gctgcacctg agcggagtgg gaatcgccct gctgatctgc
     1681 ctgttcctgg acaagcagta catcaacatc tttctgagca ggctgcccat cttctccagc
     1741 tacaatgccc agagcgagga acggcggatc atcatcagat ccttcggtat caacagcatc
     1801 aagctgccca aggaccggat ccacagcgag aagtccaaca agagcgtggc catggatatg
     1861 ctcaacgaag tgaagcggtg ccccgacgag ctgttcacaa cactgtctgc cgagaagcag
     1921 tcccggttca gaatcatcag cgacgaccac aatgaagtgc tgatgaagcg gagcagcgac
     1981 agattcgtgc ctctgctgct gcagtatatc gattacggca agctgttcga ccacatcagg
     2041 ttccacgtga acatgggcaa gctgagatac ctgctgaagg ccgacaagac ctgcatcgac
     2101 ggccagacca gagtcagagt gatcgagcag cccctgaacg gcttcggcag actggaagag
     2161 gccgagacaa tgcggaagca agagaacggc accttcggca acagcggcat ccggatcaga
     2221 gacttcgaga acatgaagcg ggacgacgcc aatcctgcca actatcccta catcgtggac
     2281 acctacacac actacatcct ggaaaacaac aaggtcgaga tgtttatcaa cgacaaagag
     2341 gacagcgccc cactgctgcc cgtgatcgag gatgatagat acgtggtcaa gacaatcccc
     2401 agctgccgga tgagcaccct ggaaattcca gccatggcct tccacatgtt tctgttcggc
     2461 agcaagaaaa ccgagaagct gatcgtggac gtgcacaacc ggtacaagag actgttccag
     2521 gccatgcaga aagaagaagt gaccgccgag aatatcgcca gcttcggaat cgccgagagc
     2581 gacctgcctc agaagatcct ggatctgatc agcggcaatg cccacggcaa ggatgtggac
     2641 gccttcatca gactgaccgt ggacgacatg ctgaccgaca ccgagcggag aatcaagaga
     2701 ttcaaggacg accggaagtc cattcggagc gccgacaaca agatgggaaa gagaggcttc
     2761 aagcagatct ccacaggcaa gctggccgac ttcctggcca aggacatcgt gctgtttcag
     2821 cccagcgtga acgatggcga gaacaagatc accggcctga actaccggat catgcagagc
     2881 gccattgccg tgtacgatag cggcgacgat tacgaggcca agcagcagtt caagctgatg
     2941 ttcgagaagg cccggctgat cggcaagggc acaacagagc ctcatccatt tctgtacaag
     3001 gtgttcgccc gcagcatccc cgccaatgcc gtcgagttct acgagcgcta cctgatcgag
     3061 cggaagttct acctgaccgg cctgtccaac gagatcaaga aaggcaacag agtggatgtg
     3121 cccttcatcc ggcgggacca gaacaagtgg aaaacacccg ccatgaagac cctgggcaga
     3181 atctacagcg aggatctgcc cgtggaactg cccagacaga tgttcgacaa tgagatcaag
     3241 tcccacctga agtccctgcc acagatggaa ggcatcgact tcaacaatgc caacgtgacc
     3301 tatctgatcg ccgagtacat gaagagagtg ctggacgacg acttccagac cttctaccag
     3361 tggaaccgca actaccggta catggacatg cttaagggcg agtacgacag aaagggctcc
     3421 ctgcagcact gcttcaccag cgtggaagag agagaaggcc tctggaaaga gcgggcctcc
     3481 agaacagagc ggtacagaaa gcaggccagc aacaagatcc gcagcaaccg gcagatgaga
     3541 aacgccagca gcgaagagat cgagacaatc ctggataagc ggctgagcaa cagccggaac
     3601 gagtaccaga aaagcgagaa agtgatccgg cgctacagag tgcaggatgc cctgctgttt
     3661 ctgctggcca aaaagaccct gaccgaactg gccgatttcg acggcgagag gttcaaactg
     3721 aaagaaatca tgcccgacgc cgagaaggga atcctgagcg agatcatgcc catgagcttc
     3781 accttcgaga aaggcggcaa gaagtacacc atcaccagcg agggcatgaa gctgaagaac
     3841 tacggcgact tctttgtgct ggctagcgac aagaggatcg gcaacctgct ggaactcgtg
     3901 ggcagcgaca tcgtgtccaa agaggatggc ggtagtggag gaagcggggg gagcggaggg
     3961 agtggtggat ccggcggctc cgatgcaatg tttgggcaac caggagccct aggtagcact
     4021 ccatttcttg gtcagcatgg ttttaatttc tttcccagtg ggattgactt ctcagcatgg
     4081 ggaaataaca gttctcaggg acagtctact cagagctctg gatatagtag caattatgct
     4141 tatgcaccta gctccttagg tggagccatg attgatggac agtcagcttt tgccaatgag
     4201 accctcaata aggctcctgg catgaatact atagaccaag ggatggcagc aggatccgat
     4261 tacaaggacg acgatgacaa gtagtgttta aacgaattcg ggctcggtac acggggatcc
     4321 tctagagtcg acctgcaggc atgcaagctg atccggctgc taacaaagcc cgaaaggaag
     4381 ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg gcggccgctt
     4441 cgagcagaca tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga
     4501 aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc
     4561 tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag
     4621 atgtgggagg tttttttaag caagtaaaac ctctacaaat gtggtaaaat cgaattctaa
     4681 tggatcctct ttgcgcttgc gttttccctt gtccagatag cccagtagct gacattcatc
     4741 cggggtcagc accgtttctg cggactggct ttctacgtgt tccgcttcct ttagcagccc
     4801 ttgcgccctg agtgcttgcg gcagcgtgag cttcaaaaga attgccagct ggggcgccct
     4861 ctggtaaggt tgggaagccc tgcaaagtaa actggatggc tttcttgccg ccaaggatct
     4921 gatggcgcag gggatcaaga tctgatcaag agacaggatg acggtcgttt cgcatgcttg
     4981 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     5041 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     5101 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg
     5161 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     5221 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     5281 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     5341 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     5401 gagcacgcac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     5461 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgtatgccg gatggtgagg
     5521 atctcgtcgt gactcatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     5581 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     5641 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     5701 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     5761 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca accggtatca
     5821 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     5881 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     5941 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     6001 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     6061 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     6121 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     6181 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     6241 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     6301 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     6361 aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc
     6421 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     6481 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatttcaaga agatcctttg
     6541 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     6601 atgagattat caaaaaggat cttcacctag atccttttat agtccggaaa tacaggaacg
     6661 cacgctggat ggcccttcgc tgggatggtg aaaccatgaa aaatggcagc ttcagtggat
     6721 taagtggggg taatgtggcc tgtaccctct ggttgcatag gtattcatac ggttaaaatt
     6781 tatcaggcgc gattgcggca gtttttcggg tggtttgttg ccatttttac ctgtctgctg
     6841 ccgtgatcgc gctgaacgcg ttttagcggt gcgtacaatt aagggattat ggtaaatcca
     6901 cttactgtct gccctcgtag ccatcgagat aaaccgcagt actccggcca cgatgcgtcc
     6961 ggcgtagagg atcgagatct tcaatattgg ccattagcca tattattcat tggttatata
     7021 gcataaatca atattggcta ttggccattg cat
//
