LOCUS       sequence-331287-        4976 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express Mito (Su9)-mRFP in zebrafish.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4976)
  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 4976)
  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-Mito-mRFP
FEATURES             Location/Qualifiers
     source          1..4976
                     /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             321..998
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     CDS             321..998
                     /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(451..469)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     719..738
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     913..936
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    1028..1162
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1052..1071)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1106..1125
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1275..1295)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1275..1293)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1314..1330)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1314..1330)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1327..1349)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1338..1354
                     /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(1362..1368,1369..1386,1387..1392))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1407..1428
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1545..1562)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1716..2304)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1796..1815)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2475..3266,3267..3335))
                     /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     3098..3117
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3336..3440)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3466..3921)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3603..3624)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3815..3834
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3992..4976
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagg atctcgagct caagcttcga attcgccatg gcctccactc
      121 gtgtcctcgc ctctcgcctg gcctcccaga tggctgcttc cgccaaggtt gcccgccctg
      181 ctgtccgcgt tgctcaggtc agcaagcgca ccatccagac tggctccccc ctccagaccc
      241 tcaagcgcac ccagatgacc tccatcgtca acgccaccac ccgccaggct ttccagaagc
      301 gcgcctactc ttcggatcca atggcctcct ccgaggacgt catcaaggag ttcatgcgct
      361 tcaaggtgcg catggagggc tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg
      421 agggccgccc ctacgagggc acccagaccg ccaagctgaa ggtgaccaag ggcggccccc
      481 tgcccttcgc ctgggacatc ctgtcccctc agttccagta cggctccaag gcctacgtga
      541 agcaccccgc cgacatcccc gactacttga agctgtcctt ccccgagggc ttcaagtggg
      601 agcgcgtgat gaacttcgag gacggcggcg tggtgaccgt gacccaggac tcctccctgc
      661 aggacggcga gttcatctac aaggtgaagc tgcgcggcac caacttcccc tccgacggcc
      721 ccgtaatgca gaagaagacc atgggctggg aggcctccac cgagcggatg taccccgagg
      781 acggcgccct gaagggcgag atcaagatga ggctgaagct gaaggacggc ggccactacg
      841 acgccgaggt caagaccacc tacatggcca agaagcccgt gcagctgccc ggcgcctaca
      901 agaccgacat caagctggac atcacctccc acaacgagga ctacaccatc gtggaacagt
      961 acgagcgcgc cgagggccgc cactccaccg gcgcctaatc tagaactata gtgagtcgta
     1021 ttacgtagat ccagacatga taagatacat tgatgagttt ggacaaacca caactagaat
     1081 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     1141 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
     1201 gggggaggtg tgggaggttt tttaattcgc ggccgcggcg ccaatgcatt gggcccggta
     1261 cccagctttt gttcccttta gtgagggtta attgcgcgct tggcgtaatc atggtcatag
     1321 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccgggagc
     1381 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     1441 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     1501 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     1561 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     1621 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     1681 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     1741 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     1801 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     1861 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     1921 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     1981 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     2041 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     2101 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     2161 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     2221 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     2281 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     2341 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     2401 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     2461 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     2521 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     2581 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     2641 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     2701 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     2761 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     2821 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     2881 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     2941 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     3001 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     3061 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     3121 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     3181 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     3241 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     3301 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     3361 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     3421 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta
     3481 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
     3541 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
     3601 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
     3661 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
     3721 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
     3781 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
     3841 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
     3901 atgcgccgct acagggcgcg tcccattcgc cattcaggct gcgcaactgt tgggaagggc
     3961 gatcggtgcg ggcctcttcg ctattacgcc agtcgaccgc caattcaata tggcgtatat
     4021 ggactcatgc caattcaata tggtggatct ggacctgtgc caattcaata tggcgtatat
     4081 ggactcgtgc caattcaata tggtggatct ggaccccagc caattcaata tggcggactt
     4141 ggcaccatgc caattcaata tggcggacct ggcactgtgc caactgggga ggggtctact
     4201 tggcacggtg ccaagtttga ggaggggtct tggccctgtg ccaagtccgc catattgaat
     4261 tggcatggtg ccaataatgg cggccatatt ggctatatgc caggatcaat atataggcaa
     4321 tatccaatat ggccctatgc caatatggct attggccagg ttcaatacta tgtattggcc
     4381 ctatgccata tagtattcca tatatgggtt ttcctattga cgtagatagc ccctcccaat
     4441 gggcggtccc atataccata tatggggctt cctaataccg cccatagcca ctcccccatt
     4501 gacgtcaatg gtctctatat atggtctttc ctattgacgt catatgggcg gtcctattga
     4561 cgtatatggc gcctccccca ttgacgtcaa ttacggtaaa tggcccgcct ggctcaatgc
     4621 ccattgacgt caataggacc acccaccatt gacgtcaatg ggatggctca ttgcccattc
     4681 atatccgttc tcacgccccc tattgacgtc aatgacggta aatggcccac ttggcagtac
     4741 atcaatatct attaatagta acttggcaag tacattacta ttggaagtac gccagggtac
     4801 attggcagta ctcccattga cgtcaatggc ggtaaatggc ccgcgatggc tgccaagtac
     4861 atccccattg acgtcaatgg ggaggggcaa tgacgcaaat gggcgttcca ttgacgtaaa
     4921 tgggcggtag gcgtgcctaa tgggaggtct atataagcaa tgctcgttta gggaac
//
