LOCUS       sequence-268883-        3944 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of mCherry mRNA for microinjection into S. purpuratus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3944)
  AUTHORS   Uchida A, Yajima M
  TITLE     An optogenetic approach to control protein localization during 
            embryogenesis of the sea urchin.
  JOURNAL   Dev Biol. 2018 Sep 1;441(1):19-30. doi: 10.1016/j.ydbio.2018.06.015.
            Epub 2018 Jun 26.
  PUBMED    29958898
REFERENCE   2  (bases 1 to 3944)
  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: 
            "10.1016/j.ydbio.2018.06.015"; journalName: "Dev Biol"; date: 
            "2018-09-1- 1"; volume: "441"; issue: "1"; pages: "19-30"
COMMENT     Sequence Label: pSP64-mCherry
FEATURES             Location/Qualifiers
     source          1..3944
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..19
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1..18
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     5'UTR           31..73
                     /label=Xenopus globin 5'-UTR
                     /note="translational enhancer from the 5'-UTR of the major 
                     beta-globin gene of Xenopus laevis"
     CDS             104..814
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTTGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(249..267)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     517..536
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     3'UTR           850..975
                     /label=Xenopus globin 3'-UTR
                     /note="3'-UTR of the major beta-globin gene of Xenopus 
                     laevis"
     primer_bind     complement(1031..1047)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1031..1047)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1044..1066)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1055..1071
                     /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(1079..1085,1086..1103,1104..1109))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1124..1145
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1262..1279)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1433..2021)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1513..1532)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2192..2983,2984..3052))
                     /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     2815..2834
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3053..3157)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3225..3243
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3281..3303)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3403..3422
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3636..3655
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 atttaggtga cactatagaa tacaagcttg cttgttcttt ttgcagaagc tcagaataaa
       61 cgctcaactt tggcagatct gatcatgcca tggttcgggc cctatggtga gcaagggcga
      121 ggaggataac atggccatca tcaaggagtt catgcgcttc aaggtgcaca tggagggctc
      181 cgtgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac
      241 ccagaccgcc aagctgaagg tgaccaaggg tggccccctg cccttcgcct gggacatcct
      301 gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag caccccgccg acatccccga
      361 ctacttgaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga
      421 cggcggcgtg gtgaccgtga cccaggactc ctccctgcag gacggcgagt tcatctacaa
      481 ggtgaagctg cgcggcacca acttcccctc cgacggcccc gtaatgcaga agaagaccac
      541 gggctgggag gcctcctccg agcggatgta ccccgaggac ggcgccctga agggcgagat
      601 caagcagagg ctgaagctga aggacggcgg ccactacgac gctgaggtca agaccaccta
      661 caaggccaag aagcccgtgc agctgcccgg cgcctacaac gtcaacatca agttggacat
      721 cacctcccac aacgaggact acaccatcgt ggaacagtac gaacgcgccg agggccgcca
      781 ctccaccggc ggcatggacg agctgtacaa gtaataaact agtgactgac taggatctgg
      841 ttaccactaa accagcctca agaacacccg aatggagtct ctaagctaca taataccaac
      901 ttacacttta caaaatgttg tcccccaaaa tgtagccatt cgtatctgct cctaataaaa
      961 agaaagtttc ttcacccccc ctgcaggtcg actctagagg atccccgggc gagctcgaat
     1021 tcgtaatcat gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     1081 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     1141 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     1201 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     1261 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     1321 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     1381 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     1441 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     1501 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     1561 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     1621 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     1681 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     1741 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     1801 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     1861 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     1921 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     1981 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     2041 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     2101 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     2161 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     2221 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     2281 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     2341 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     2401 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     2461 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     2521 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     2581 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     2641 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     2701 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     2761 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     2821 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     2881 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     2941 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     3001 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     3061 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     3121 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     3181 ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga
     3241 ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc
     3301 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
     3361 cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg
     3421 tactgagagt gcaccattcg acgctctccc ttatgcgact cctgcattag gaagcagccc
     3481 agtagtaggt tgaggccgtt gagcaccgcc gccgcaagga atggtgcatg caaggagatg
     3541 gcgcccaaca gtcccccggc cacggggcct gccaccatac ccacgccgaa acaagcgctc
     3601 atgagcccga agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca
     3661 gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta gaggatctgg
     3721 ctagcgatga ccctgctgat tggttcgctg accatttccg ggtgcgggac ggcgttacca
     3781 gaaactcaga aggttcgtcc aaccaaaccg actctgacgg cagtttacga gagagatgat
     3841 agggtctgct tcagtaagcc agatgctaca caattaggct tgtacatatt gtcgttagaa
     3901 cgcggctaca attaatacat aaccttatgt atcatacaca tacg
//
