LOCUS       sequence-327711-        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"
     promoter        complement(1..105)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             467..790
                     /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"
     polyA_signal    1324..1405
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1361..1380)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1416..1433)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1587..2175)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1667..1686)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2346..3137,3138..3206))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2969..2988
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
       61 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
      121 agtgccacct gacgtacgtg gactgggcat agtagagcac acctttagcc tcagcacttg
      181 ggaggcagag gcaagtagat ctctgagttc aaggttagcc tggtctacat ggagagttag
      241 ttccaggaca gcctgagata ggtatcacca gaaagtagaa cattttaagt attaagccct
      301 tacaggattt ttggtttgcc tcttctttcc cccttgtttt aggattactc ttggaaacca
      361 actggatggg atacacctac tagacccaga tgtggttgcc aggtttaagc aaaggtatcc
      421 ggacggcatc atctctaaac ctaagaattt aggtggaagt ggaggcggag tgcaggtgga
      481 aaccatctcc ccaggagacg ggcgcacctt ccccaagcgc ggccagacct gcgtggtgca
      541 ctacaccggg atgcttgaag atggaaagaa agttgattcc tcccgggaca gaaacaagcc
      601 ctttaagttt atgctaggca agcaggaggt gatccgaggc tgggaagaag gggttgccca
      661 gatgagtgtg ggtcagagag ccaaactgac tatatctcca gattatgcct atggtgccac
      721 tgggcaccca ggcatcatcc caccacatgc cactctcgtc ttcgatgtgg agcttctaaa
      781 actggaatag acgcacttcc ttacaaagct aagctttcag agagatgggc tacaggccac
      841 accttaagag gactgtttac gcaatagtgt cccctaccct gtatataaat aaaagtttgt
      901 attataattg gaatttctga agtgaaatta tggctttcca cttagtgtgt gtccttgtgc
      961 tactcttatt tagcgtgatt cattataagt aaaagaagaa tgttagctac ctagcgtgtt
     1021 agaaaagact acacaggttc acagtgctta gcaattagta tgctataaac aggattaatt
     1081 tgaattttac catgcatatg gtactctaaa cttttgctta ttctatgtac gtttgctggt
     1141 ttattagtct gtactctagt cagctgacgc gtgctagcgc ggccgcatcg ataagcttgt
     1201 cgacgatatc tctagaggat cataatcagc cataccacat ttgtagaggt tttacttgct
     1261 ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt
     1321 gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     1381 acaaataaag catttttttc actgcctcga gcttcctcgc tcactgactc gctgcgctcg
     1441 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     1501 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     1561 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     1621 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     1681 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     1741 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     1801 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     1861 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     1921 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     1981 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     2041 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     2101 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     2161 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     2221 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     2281 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     2341 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     2401 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     2461 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     2521 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     2581 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     2641 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     2701 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     2761 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     2821 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     2881 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     2941 agttgctctt gcccggcgtc aacacgggat aataccgcgc cacatagcag aactttaaaa
     3001 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     3061 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     3121 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     3181 gcgacacgga aatgttgaat actcat
//
