LOCUS       sequence-321550-        3206 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Template for knockin FBKP-V into Mettl3 C-terminus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3206)
  AUTHORS   Wei G, Almeida M, Pintacuda G, Coker H, Bowness JS, Ule J, 
            Brockdorff N
  TITLE     Acute depletion of METTL3 implicates N (6)-methyladenosine in 
            alternative intron/exon inclusion in the nascent transcriptome.
  JOURNAL   Genome Res. 2021 Aug;31(8):1395-1408. doi: 10.1101/gr.271635.120. 
            Epub 2021 Jun 15.
  PUBMED    34131006
REFERENCE   2  (bases 1 to 3206)
  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: 
            "10.1101/gr.271635.120"; journalName: "Genome Res"; date: "2021-08";
            volume: "31"; issue: "8"; pages: "1395-1408"
COMMENT     Sequence Label: pTargeting_Mettl3C_FKBP-V
FEATURES             Location/Qualifiers
     source          1..3206
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    171..252
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(208..227)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(263..280)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(434..1022)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(514..533)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1193..1984,1985..2053))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     1816..1835
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2054..2158)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2520..2843
                     /codon_start=1
                     /product="F36V mutant of FK506-binding protein FKBP12"
                     /label=FKBP-V
                     /note="binds synthetic ligands that do not bind wild-type 
                     FKBP"
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKVDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
ORIGIN
        1 ctctagtcag ctgacgcgtg ctagcgcggc cgcatcgata agcttgtcga cgatatctct
       61 agaggatcat aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc
      121 cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta
      181 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
      241 ttttttcact gcctcgagct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
      301 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
      361 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
      421 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
      481 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
      541 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
      601 tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc agttcggtgt
      661 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
      721 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
      781 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
      841 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
      901 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
      961 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     1021 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     1081 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     1141 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     1201 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     1261 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     1321 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     1381 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     1441 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     1501 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     1561 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     1621 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     1681 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     1741 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     1801 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     1861 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     1921 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     1981 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     2041 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     2101 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     2161 catttccccg aaaagtgcca cctgacgtac gtggactggg catagtagag cacaccttta
     2221 gcctcagcac ttgggaggca gaggcaagta gatctctgag ttcaaggtta gcctggtcta
     2281 catggagagt tagttccagg acagcctgag ataggtatca ccagaaagta gaacatttta
     2341 agtattaagc ccttacagga tttttggttt gcctcttctt tcccccttgt tttaggatta
     2401 ctcttggaaa ccaactggat gggatacacc tactagaccc agatgtggtt gccaggttta
     2461 agcaaaggta tccggacggc atcatctcta aacctaagaa tttaggtgga agtggaggcg
     2521 gagtgcaggt ggaaaccatc tccccaggag acgggcgcac cttccccaag cgcggccaga
     2581 cctgcgtggt gcactacacc gggatgcttg aagatggaaa gaaagttgat tcctcccggg
     2641 acagaaacaa gccctttaag tttatgctag gcaagcagga ggtgatccga ggctgggaag
     2701 aaggggttgc ccagatgagt gtgggtcaga gagccaaact gactatatct ccagattatg
     2761 cctatggtgc cactgggcac ccaggcatca tcccaccaca tgccactctc gtcttcgatg
     2821 tggagcttct aaaactggaa tagacgcact tccttacaaa gctaagcttt cagagagatg
     2881 ggctacaggc cacaccttaa gaggactgtt tacgcaatag tgtcccctac cctgtatata
     2941 aataaaagtt tgtattataa ttggaatttc tgaagtgaaa ttatggcttt ccacttagtg
     3001 tgtgtccttg tgctactctt atttagcgtg attcattata agtaaaagaa gaatgttagc
     3061 tacctagcgt gttagaaaag actacacagg ttcacagtgc ttagcaatta gtatgctata
     3121 aacaggatta atttgaattt taccatgcat atggtactct aaacttttgc ttattctatg
     3181 tacgtttgct ggtttattag tctgta
//
