LOCUS       sequence-331460-        5724 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express zebrafish Dnase1l1l-mRFP in zebrafish.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5724)
  AUTHORS   Morishita H, Eguchi T, Tsukamoto S, Sakamaki Y, Takahashi S, Saito 
            C, Koyama-Honda I, Mizushima N
  TITLE     Organelle degradation in the lens by PLAAT phospholipases.
  JOURNAL   Nature. 2021 Apr;592(7855):634-638. doi: 10.1038/s41586-021-03439-w.
            Epub 2021 Apr 14.
  PUBMED    33854238
REFERENCE   2  (bases 1 to 5724)
  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: 
            "10.1038/s41586-021-03439-w"; journalName: "Nature"; date: 
            "2021-04"; volume: "592"; issue: "7855"; pages: "634-638"
COMMENT     Sequence Label: pCS2-Dnase1l1l-mRFP
FEATURES             Location/Qualifiers
     source          1..5724
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        999..1017
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     999..1016
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     misc_feature    1047..1099
                     /label=MCS
                     /note="multiple cloning site"
     CDS             2033..2710
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     CDS             2033..2710
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /note="mammalian codon-optimized"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     primer_bind     complement(2163..2181)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2431..2450
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2625..2648
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    2740..2874
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2764..2783)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2818..2837
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2987..3007)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2987..3005)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3026..3042)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3026..3042)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3039..3061)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3050..3066
                     /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(3074..3080,3081..3098,3099..3104))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3119..3140
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3257..3274)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3428..4016)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3508..3527)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4187..4978,4979..5047))
                     /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     4810..4829
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5048..5152)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5178..5633)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5315..5336)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5527..5546
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        join(5711..5724,1..964)
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 gcgtatatgg actcatgcca attcaatatg gtggatctgg acctgtgcca attcaatatg
       61 gcgtatatgg actcgtgcca attcaatatg gtggatctgg accccagcca attcaatatg
      121 gcggacttgg caccatgcca attcaatatg gcggacctgg cactgtgcca actggggagg
      181 ggtctacttg gcacggtgcc aagtttgagg aggggtcttg gccctgtgcc aagtccgcca
      241 tattgaattg gcatggtgcc aataatggcg gccatattgg ctatatgcca ggatcaatat
      301 ataggcaata tccaatatgg ccctatgcca atatggctat tggccaggtt caatactatg
      361 tattggccct atgccatata gtattccata tatgggtttt cctattgacg tagatagccc
      421 ctcccaatgg gcggtcccat ataccatata tggggcttcc taataccgcc catagccact
      481 cccccattga cgtcaatggt ctctatatat ggtctttcct attgacgtca tatgggcggt
      541 cctattgacg tatatggcgc ctcccccatt gacgtcaatt acggtaaatg gcccgcctgg
      601 ctcaatgccc attgacgtca ataggaccac ccaccattga cgtcaatggg atggctcatt
      661 gcccattcat atccgttctc acgcccccta ttgacgtcaa tgacggtaaa tggcccactt
      721 ggcagtacat caatatctat taatagtaac ttggcaagta cattactatt ggaagtacgc
      781 cagggtacat tggcagtact cccattgacg tcaatggcgg taaatggccc gcgatggctg
      841 ccaagtacat ccccattgac gtcaatgggg aggggcaatg acgcaaatgg gcgttccatt
      901 gacgtaaatg ggcggtaggc gtgcctaatg ggaggtctat ataagcaatg ctcgtttagg
      961 gaaccgccat tctgcctggg gacgtcggag caagcttgat ttaggtgaca ctatagaata
     1021 caagctactt gttctttttg caggatctcg agctcaagct tcgaattctg cagtcgacgg
     1081 taccgcgggc ccgggatcca ctagtaacgg ccgccagtgt gctggaattc gcccttggga
     1141 attcaccatg agccctccat tgctcctgtg ttttattgga ctgtgtttgt ggacttgtgt
     1201 aatttccctg aagatctgcg ccttcaacat ccaaagctat ggagaagcaa aggccagcaa
     1261 tactagagtt atggagattt tgacaaagat tatttctcgc tgtgacctga gtttgattca
     1321 agaggttcga gattctagag gagaagccat ttctgcactg ttgatgagtc tgaacagatt
     1381 cgacaaatct catatttatg cccatttaga gagtaaaaga ttggggaaga aaacatacaa
     1441 agagcagtat gtgtatattt acagaaaaga catgctacag gtgcaagagc aatacgagca
     1501 cccagaggtc aatgaaagtg catcagaagt cttttccaga gagcccttca tcatactggt
     1561 ccactctcca acaaccttgg tgaagaattt tgtcctgatt ggtcagcaca cctctcctaa
     1621 gagtgcaatg aaagaaatgg attcactgta cgaggtcttt caaaccgtca agaagaaatg
     1681 gaaaactgag aatgtgatgt ttctgggaga cttaaatgcc gcctgcaact acgtcactaa
     1741 taaaggactg agaaatgtgc gtctcaggag tgacccaaag ttccactggt tgatcagaga
     1801 cgaacaagac actacagtgc gtgaaaaaac acgctgcgct tatgacagga ttattattca
     1861 tggaaaagag ctcatttctg ggatagttcc ggaatccgct caaccgttca acttcaaaca
     1921 ggaatttaat ctctctgagg aagaggcttt ggaagtcagt gaccactttc cagtagaagt
     1981 ggatttaaaa gcaaaccatc gctacctcct gcgccatgaa ctgctggatc cgatggcctc
     2041 ctccgaggac gtcatcaagg agttcatgcg cttcaaggtg cgcatggagg gctccgtgaa
     2101 cggccacgag ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac
     2161 cgccaagctg aaggtgacca agggcggccc cctgcccttc gcctgggaca tcctgtcccc
     2221 tcagttccag tacggctcca aggcctacgt gaagcacccc gccgacatcc ccgactactt
     2281 gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg
     2341 cgtggtgacc gtgacccagg actcctccct gcaggacggc gagttcatct acaaggtgaa
     2401 gctgcgcggc accaacttcc cctccgacgg ccccgtaatg cagaagaaga ccatgggctg
     2461 ggaggcctcc accgagcgga tgtaccccga ggacggcgcc ctgaagggcg agatcaagat
     2521 gaggctgaag ctgaaggacg gcggccacta cgacgccgag gtcaagacca cctacatggc
     2581 caagaagccc gtgcagctgc ccggcgccta caagaccgac atcaagctgg acatcacctc
     2641 ccacaacgag gactacacca tcgtggaaca gtacgagcgc gccgagggcc gccactccac
     2701 cggcgcctag tctagaacta tagtgagtcg tattacgtag atccagacat gataagatac
     2761 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
     2821 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
     2881 aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaattc
     2941 gcggccgcgg cgccaatgca ttgggcccgg tacccagctt ttgttccctt tagtgagggt
     3001 taattgcgcg cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     3061 tcacaattcc acacaacata cgagccggga gcataaagtg taaagcctgg ggtgcctaat
     3121 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     3181 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     3241 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     3301 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     3361 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     3421 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     3481 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     3541 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     3601 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     3661 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     3721 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     3781 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     3841 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     3901 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     3961 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     4021 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     4081 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     4141 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     4201 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     4261 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     4321 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     4381 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     4441 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     4501 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     4561 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     4621 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     4681 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     4741 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     4801 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     4861 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     4921 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     4981 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     5041 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     5101 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     5161 cccgaaaagt gccacctaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt
     5221 ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc
     5281 aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt
     5341 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact
     5401 acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg
     5461 gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag
     5521 aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac
     5581 gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcccattc
     5641 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
     5701 ccagtcgacc gccaattcaa tatg
//
