LOCUS       sequence-355699-        5180 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses zebrafish ythdf3 with 3x flag tag, for mRNA injection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5180)
  AUTHORS   Kontur C, Jeong M, Cifuentes D, Giraldez AJ
  TITLE     Ythdf m(6)A Readers Function Redundantly during Zebrafish 
            Development.
  JOURNAL   Cell Rep. 2020 Dec 29;33(13):108598. doi: 
            10.1016/j.celrep.2020.108598.
  PUBMED    33378672
REFERENCE   2  (bases 1 to 5180)
  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"; journalName: "Cell Rep.";
            date: "2020-12-29"; volume: "33(13):108598. doi"; pages: " 
            10.1016/j.celrep.2020.108598"
COMMENT     Sequence Label: pSP64T-ythdf3-3xflag
FEATURES             Location/Qualifiers
     source          1..5180
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(11..28)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(182..770)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(262..281)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(941..1732,1733..1801))
                     /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     1564..1583
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1802..1906)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1974..1992
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2030..2052)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2152..2171
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2385..2404
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        2694..2712
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     2694..2711
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     5'UTR           2724..2766
                     /label=Xenopus globin 5'-UTR
                     /note="translational enhancer from the 5'-UTR of the major 
                     beta-globin gene of Xenopus laevis"
     regulatory      2777..2786
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2786..2851
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     3'UTR           4707..4832
                     /label=Xenopus globin 3'-UTR
                     /note="3'-UTR of the major beta-globin gene of Xenopus 
                     laevis"
     primer_bind     complement(4960..4976)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4960..4976)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4973..4995)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4984..5000
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5008..5014,5015..5032,5033..5038))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5053..5074
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
       61 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
      121 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
      181 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
      241 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
      301 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
      361 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
      421 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
      481 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
      541 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
      601 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
      661 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
      721 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
      781 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
      841 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
      901 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
      961 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     1021 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     1081 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     1141 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     1201 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     1261 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     1321 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     1381 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     1441 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     1501 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     1561 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     1621 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     1681 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     1741 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     1801 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     1861 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     1921 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     1981 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     2041 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     2101 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag
     2161 agcagattgt actgagagtg caccattcga cgctctccct tatgcgactc ctgcattagg
     2221 aagcagccca gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc
     2281 aaggagatgg cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa
     2341 caagcgctca tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata
     2401 taggcgccag caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag
     2461 aggatctggc tagcgatgac cctgctgatt ggttcgctga ccatttccgg gtgcgggacg
     2521 gcgttaccag aaactcagaa ggttcgtcca accaaaccga ctctgacggc agtttacgag
     2581 agagatgata gggtctgctt cagtaagcca gatgctacac aattaggctt gtacatattg
     2641 tcgttagaac gcggctacaa ttaatacata accttatgta tcatacacat acgatttagg
     2701 tgacactata gaatacaagc ttgcttgttc tttttgcaga agctcagaat aaacgctcaa
     2761 ctttggcaga tccgcggccg ccatggacta caaagaccat gacggtgatt ataaagatca
     2821 tgacatcgac tacaaggatg acgatgacaa gaattcaggt tctggctcag gatcgtctgc
     2881 gacaactgtc gatcagagac ctaaaggaca aggaaacaaa gtgcaaaacg gttcgatgca
     2941 tcagaaggat gctgtaaatg atgatgactt tgacaattac ctgagcagtc agtcaaatca
     3001 gagtaatagc tacccaccaa tgtctgatcc ctacatgccc agctactatg ctccctctat
     3061 tggattccct tactccctgg gagaggctgc ttggtccact gcaggagacc ctcctatgcc
     3121 ttacctgacc acgtatggac agatgagcaa tggtgagcat cacttcattc ctgacggagt
     3181 cttcagccag cccggtgctt taggcaacac gccccccttt ctcagtcaac acggatttaa
     3241 cttctttcct ggaaatgcag acttttctac ctggggcacc agtggttctc agggacagtc
     3301 aacacagagt tcagcgtaca gcagtagtta tggatacccg ccaagctctt tgggccgggc
     3361 cattgctgat ggacaggctg gatttggtag tgacacgcaa cttagtaaag ttccagggct
     3421 cagtagtatc gatcagggaa tggcaggact gaagctcggc tctgacatgt cagctgttac
     3481 gaaaactgtt gggtctcctt taggagggac ggtgggtatg agtagcatgg ccgctaacag
     3541 catgccacct gtcagctcct ctgcaccgaa acccacttca tgggcagcca tcgcgaggaa
     3601 acctgcaaag cctcagccta agatgaagcc gaagcccaac atgattttgg gaagcggcgt
     3661 cacaattcct cctccgccaa taaagcataa cataaacatt gggacttggg aggacaaagg
     3721 gtccatcacc aaaccccctc ttgcgcagca gatgctgccc cctcagcagt tggtacagca
     3781 gcaactcttg gcccaacctc cacccatgct tcaaagtcct ttgccacctc aacaccaacc
     3841 ccaacaccag cagctccagg tccaatctcc ccagcaccct cagcatcttc ccccaggtcc
     3901 tccccaccat cactcgcagc caggacctcc acagcccctg cacccatctc aggcgcagaa
     3961 ccccctaatc caaaaccgct gggtggcccc tcggaaccga ggaaccatgt ttaaccagaa
     4021 cagcggcatg gataactttg gccttgggac aggagtcccc atgagctcat cgccttcctc
     4081 cagcgaggtt catcccgtgc tggaaaagct taaggccctc aacaattaca accccaaaga
     4141 cttcgactgg accttgaaaa acggacgggt cttcatcatc aagagctact ctgaggacga
     4201 catccaccgc tccatcaagt actccatctg gtgcagcaca gagcatggaa acaagcgcct
     4261 ggatggtgcc taccgctcgc tcagtgccaa aggtccactg tacctgctct ttagtgtcaa
     4321 tggcagtgga cacttctgcg gcgtggcgga gatgaagtca accgtggact ataacgcgta
     4381 tgctggcgtc tggtcccagg ataaatggaa gggcaagttt gaggtgaaat ggatctttgt
     4441 taaggacgtg cccaataacc agctccggca catccgctta gaaaacaatg acaacaagcc
     4501 tgtcaccaac tccagagaca ctcaggaagt gcccctagag aaagcaaagc aagtgcttaa
     4561 aattattgcg actttcaagc ataccacctc aatctttgac gattttgcac attacgaaaa
     4621 gcggcaggag gaagaggaag ccatgcgaag ggagcgaaat agaaataagc agtagctcga
     4681 gaataattcg gatctggtta ccactaaacc agcctcaaga acacccgaat ggagtctcta
     4741 agctacataa taccaactta cactttacaa aatgttgtcc cccaaaatgt agccattcgt
     4801 atctgctcct aataaaaaga aagtttcttc acattctaaa aaaaaaaaaa aaaaaaaaaa
     4861 aaaaaaaaaa aaaaaaaaaa aaaaaaaamc cccccccccc cccccctgca ggtcgactct
     4921 agaggatccc cgggcgagct cgaattaatt cgtaatcatg tcatagctgt ttcctgtgtg
     4981 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     5041 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     5101 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     5161 cggtttgcgt attgggcgct
//
