LOCUS       sequence-355701-        4689 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Reporter to test methylated mRNA stability, for mRNA injection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4689)
  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 4689)
  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: pCS2+methylated-reporter
FEATURES             Location/Qualifiers
     source          1..4689
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             520..546
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    741..875
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(765..784)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     819..838
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(988..1008)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(988..1006)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1027..1043)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1027..1043)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1040..1062)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1051..1067
                     /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(1075..1081,1082..1099,1100..1105))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1120..1141
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1258..1275)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1429..2017)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1509..1528)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2188..2979,2980..3048))
                     /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     2811..2830
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3049..3153)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3179..3634)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3316..3337)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3528..3547
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3705..4689
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agatttcttg
       61 cttcttgttc tttttgctgg ttccatggcc cgcctttgtg ctgcccgttc ctctgctggt
      121 cgcctttggg ctgcccggct ctgggctggt gtgctctgtg ctggccggct gttgttcggc
      181 gcgtcctctg ggggcgtcct ttctgctggt ccttcctgtg ggggtgtcct gtgtggggcc
      241 gtgctttggg ctgccgtgct gtctgctggc cccctctgcg ctggtccgct ttgcgggggt
      301 cgcctgttgg ctgcccgtct ctgcgggggt cgtctgttgg ggggccctct ctgggctggc
      361 gtttcctctg ctggtccgtc cttgttcggc gttctctgtt tcggtcgcct gtgtttcggt
      421 ttttcctctt tcggtcctct gtgtgctggc tttctgtgcg ctggccgcct tttggctggc
      481 cgtctctgcg ggggccgtct cttgggggcc tcgctgtggt acccttatga cgtgcccgat
      541 tacgcttgac tcgagctgtc cttcggcgtg ctgtgggact ggactggact ggactggact
      601 ggactggact ggactggact ggactggact ggactgtccg ccttgcccgt tggctgccgc
      661 ttccccgtcc tcttgggatc ccatcgattc gaattcaagg cctctcgagc ctctagaact
      721 atagtgagtc gtattacgta gatccagaca tgataagata cattgatgag tttggacaaa
      781 ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt
      841 tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta
      901 tgtttcaggt tcagggggag gtgtgggagg ttttttaatt cgcggccgcg gcgccaatgc
      961 attgggcccg gtacccagct tttgttccct ttagtgaggg ttaattgcgc gcttggcgta
     1021 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     1081 acgagccggg agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     1141 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     1201 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     1261 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     1321 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     1381 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     1441 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     1501 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     1561 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     1621 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     1681 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     1741 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     1801 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     1861 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     1921 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     1981 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     2041 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     2101 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     2161 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     2221 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     2281 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     2341 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     2401 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     2461 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     2521 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     2581 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     2641 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     2701 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     2761 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     2821 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     2881 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     2941 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     3001 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     3061 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     3121 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctaa
     3181 attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
     3241 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
     3301 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
     3361 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa
     3421 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
     3481 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
     3541 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
     3601 cccgccgcgc ttaatgcgcc gctacagggc gcgtcccatt cgccattcag gctgcgcaac
     3661 tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagtcgac cgccaattca
     3721 atatggcgta tatggactca tgccaattca atatggtgga tctggacctg tgccaattca
     3781 atatggcgta tatggactcg tgccaattca atatggtgga tctggacccc agccaattca
     3841 atatggcgga cttggcacca tgccaattca atatggcgga cctggcactg tgccaactgg
     3901 ggaggggtct acttggcacg gtgccaagtt tgaggagggg tcttggccct gtgccaagtc
     3961 cgccatattg aattggcatg gtgccaataa tggcggccat attggctata tgccaggatc
     4021 aatatatagg caatatccaa tatggcccta tgccaatatg gctattggcc aggttcaata
     4081 ctatgtattg gccctatgcc atatagtatt ccatatatgg gttttcctat tgacgtagat
     4141 agcccctccc aatgggcggt cccatatacc atatatgggg cttcctaata ccgcccatag
     4201 ccactccccc attgacgtca atggtctcta tatatggtct ttcctattga cgtcatatgg
     4261 gcggtcctat tgacgtatat ggcgcctccc ccattgacgt caattacggt aaatggcccg
     4321 cctggctcaa tgcccattga cgtcaatagg accacccacc attgacgtca atgggatggc
     4381 tcattgccca ttcatatccg ttctcacgcc ccctattgac gtcaatgacg gtaaatggcc
     4441 cacttggcag tacatcaata tctattaata gtaacttggc aagtacatta ctattggaag
     4501 tacgccaggg tacattggca gtactcccat tgacgtcaat ggcggtaaat ggcccgcgat
     4561 ggctgccaag tacatcccca ttgacgtcaa tggggagggg caatgacgca aatgggcgtt
     4621 ccattgacgt aaatgggcgg taggcgtgcc taatgggagg tctatataag caatgctcgt
     4681 ttagggaac
//
