LOCUS       sequence-227044-        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        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..721
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     672..692
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          857..989
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     972..991
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             4331..4354
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(4472..4490)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    4527..4648
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4538..4557)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4592..4611
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             5047..5841
                     /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(5101..5120)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5711..5730
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(6013..6601)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6093..6112)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    6717..7000
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      661 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      721 tggtttagtg aaccgtcaga tcactagaag ctttattgcg gtagtttatc acagttaaat
      781 tgctaacgca gtcagtgctt ctgacacaac agtctcgaac ttaagctgca gaagttggtc
      841 gtgaggcact gggcaggtaa gtatcaaggt tacaagacag gtttaaggag accaatagaa
      901 actgggcttg tcgagacaga gaagactctt gcgtttctga taggcaccta ttggtcttac
      961 tgacatccac tttgcctttc tctccacagg tgtccactcc cagttcaatt acagctctta
     1021 aggcagatca ctagaagctt acgctaccac catgaacatc cccgctctgg tggaaaacca
     1081 gaagaagtac tttggcacct acagcgtgat ggccatgctg aacgctcaga ccgtgctgga
     1141 ccacatccag aaggtggccg atattgaggg cgagcagaac gagaacaacg agaatctgtg
     1201 gtttcacccc gtgatgagcc acctgtacaa cgccaagaac ggctacgaca agcagcccga
     1261 gaaaaccatg ttcatcatcg agcggctgca gagctacttc ccattcctga agatcatggc
     1321 cgagaaccag agagagtaca gcaacggcaa gtacaagcag aaccgcgtgg aagtgaacag
     1381 caacgacatc ttcgaggtgc tgaagcgcgc cttcggcgtg ctgaagatgt acagggacct
     1441 gaccaacgca tacaagacct acgaggaaaa gctgaacgac ggctgcgagt tcctgaccag
     1501 cacagagcaa cctctgagcg gcatgatcaa caactactac acagtggccc tgcggaacat
     1561 gaacgagaga tacggctaca agacagagga cctggccttc atccaggaca agcggttcaa
     1621 gttcgtgaag gacgcctacg gcaagaaaaa gtcccaagtg aataccggat tcttcctgag
     1681 cctgcaggac tacaacggcg acacacagaa gaagctgcac ctgagcggag tgggaatcgc
     1741 cctgctgatc tgcctgttcc tggacaagca gtacatcaac atctttctga gcaggctgcc
     1801 catcttctcc agctacaatg cccagagcga ggaacggcgg atcatcatca gatccttcgg
     1861 tatcaacagc atcaagctgc ccaaggaccg gatccacagc gagaagtcca acaagagcgt
     1921 ggccatggat atgctcaacg aagtgaagcg gtgccccgac gagctgttca caacactgtc
     1981 tgccgagaag cagtcccggt tcagaatcat cagcgacgac cacaatgaag tgctgatgaa
     2041 gcggagcagc gacagattcg tgcctctgct gctgcagtat atcgattacg gcaagctgtt
     2101 cgaccacatc aggttccacg tgaacatggg caagctgaga tacctgctga aggccgacaa
     2161 gacctgcatc gacggccaga ccagagtcag agtgatcgag cagcccctga acggcttcgg
     2221 cagactggaa gaggccgaga caatgcggaa gcaagagaac ggcaccttcg gcaacagcgg
     2281 catccggatc agagacttcg agaacatgaa gcgggacgac gccaatcctg ccaactatcc
     2341 ctacatcgtg gacacctaca cacactacat cctggaaaac aacaaggtcg agatgtttat
     2401 caacgacaaa gaggacagcg ccccactgct gcccgtgatc gaggatgata gatacgtggt
     2461 caagacaatc cccagctgcc ggatgagcac cctggaaatt ccagccatgg ccttccacat
     2521 gtttctgttc ggcagcaaga aaaccgagaa gctgatcgtg gacgtgcaca accggtacaa
     2581 gagactgttc caggccatgc agaaagaaga agtgaccgcc gagaatatcg ccagcttcgg
     2641 aatcgccgag agcgacctgc ctcagaagat cctggatctg atcagcggca atgcccacgg
     2701 caaggatgtg gacgccttca tcagactgac cgtggacgac atgctgaccg acaccgagcg
     2761 gagaatcaag agattcaagg acgaccggaa gtccattcgg agcgccgaca acaagatggg
     2821 aaagagaggc ttcaagcaga tctccacagg caagctggcc gacttcctgg ccaaggacat
     2881 cgtgctgttt cagcccagcg tgaacgatgg cgagaacaag atcaccggcc tgaactaccg
     2941 gatcatgcag agcgccattg ccgtgtacga tagcggcgac gattacgagg ccaagcagca
     3001 gttcaagctg atgttcgaga aggcccggct gatcggcaag ggcacaacag agcctcatcc
     3061 atttctgtac aaggtgttcg cccgcagcat ccccgccaat gccgtcgagt tctacgagcg
     3121 ctacctgatc gagcggaagt tctacctgac cggcctgtcc aacgagatca agaaaggcaa
     3181 cagagtggat gtgcccttca tccggcggga ccagaacaag tggaaaacac ccgccatgaa
     3241 gaccctgggc agaatctaca gcgaggatct gcccgtggaa ctgcccagac agatgttcga
     3301 caatgagatc aagtcccacc tgaagtccct gccacagatg gaaggcatcg acttcaacaa
     3361 tgccaacgtg acctatctga tcgccgagta catgaagaga gtgctggacg acgacttcca
     3421 gaccttctac cagtggaacc gcaactaccg gtacatggac atgcttaagg gcgagtacga
     3481 cagaaagggc tccctgcagc actgcttcac cagcgtggaa gagagagaag gcctctggaa
     3541 agagcgggcc tccagaacag agcggtacag aaagcaggcc agcaacaaga tccgcagcaa
     3601 ccggcagatg agaaacgcca gcagcgaaga gatcgagaca atcctggata agcggctgag
     3661 caacagccgg aacgagtacc agaaaagcga gaaagtgatc cggcgctaca gagtgcagga
     3721 tgccctgctg tttctgctgg ccaaaaagac cctgaccgaa ctggccgatt tcgacggcga
     3781 gaggttcaaa ctgaaagaaa tcatgcccga cgccgagaag ggaatcctga gcgagatcat
     3841 gcccatgagc ttcaccttcg agaaaggcgg caagaagtac accatcacca gcgagggcat
     3901 gaagctgaag aactacggcg acttctttgt gctggctagc gacaagagga tcggcaacct
     3961 gctggaactc gtgggcagcg acatcgtgtc caaagaggat ggcggtagtg gaggaagcgg
     4021 ggggagcgga gggagtggtg gatccggcgg ctccgatgca atgtttgggc aaccaggagc
     4081 cctaggtagc actccatttc ttggtcagca tggttttaat ttctttccca gtgggattga
     4141 cttctcagca tggggaaata acagttctca gggacagtct actcagagct ctggatatag
     4201 tagcaattat gcttatgcac ctagctcctt aggtggagcc atgattgatg gacagtcagc
     4261 ttttgccaat gagaccctca ataaggctcc tggcatgaat actatagacc aagggatggc
     4321 agcaggatcc gattacaagg acgacgatga caagtagtgt ttaaacgaat tcgggctcgg
     4381 tacacgggga tcctctagag tcgacctgca ggcatgcaag ctgatccggc tgctaacaaa
     4441 gcccgaaagg aagctgagtt ggctgctgcc accgctgagc aataactagc ataacccctt
     4501 ggggcggccg cttcgagcag acatgataag atacattgat gagtttggac aaaccacaac
     4561 tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt
     4621 aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca
     4681 ggttcagggg gagatgtggg aggttttttt aagcaagtaa aacctctaca aatgtggtaa
     4741 aatcgaattc taatggatcc tctttgcgct tgcgttttcc cttgtccaga tagcccagta
     4801 gctgacattc atccggggtc agcaccgttt ctgcggactg gctttctacg tgttccgctt
     4861 cctttagcag cccttgcgcc ctgagtgctt gcggcagcgt gagcttcaaa agaattgcca
     4921 gctggggcgc cctctggtaa ggttgggaag ccctgcaaag taaactggat ggctttcttg
     4981 ccgccaagga tctgatggcg caggggatca agatctgatc aagagacagg atgacggtcg
     5041 tttcgcatgc ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     5101 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     5161 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     5221 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     5281 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     5341 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     5401 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     5461 catcgcatcg agcgagcacg cactcggatg gaagccggtc ttgtcgatca ggatgatctg
     5521 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgtatg
     5581 ccggatggtg aggatctcgt cgtgactcat ggcgatgcct gcttgccgaa tatcatggtg
     5641 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     5701 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     5761 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     5821 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     5881 ccaaccggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     5941 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     6001 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     6061 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     6121 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     6181 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     6241 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     6301 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     6361 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     6421 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     6481 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     6541 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatttca
     6601 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     6661 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt tatagtccgg
     6721 aaatacagga acgcacgctg gatggccctt cgctgggatg gtgaaaccat gaaaaatggc
     6781 agcttcagtg gattaagtgg gggtaatgtg gcctgtaccc tctggttgca taggtattca
     6841 tacggttaaa atttatcagg cgcgattgcg gcagtttttc gggtggtttg ttgccatttt
     6901 tacctgtctg ctgccgtgat cgcgctgaac gcgttttagc ggtgcgtaca attaagggat
     6961 tatggtaaat ccacttactg tctgccctcg tagccatcga gataaaccgc agtactccgg
     7021 ccacgatgcg tccggcgtag aggatcgaga tct
//
