LOCUS       sequence-269591-        4178 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of membrane-mCherry mRNA for microinjection for 
            visualization of membrane in S. pupuratus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4178)
  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 4178)
  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-membrane-mCherry
FEATURES             Location/Qualifiers
     source          1..4178
                     /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             166..873
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(311..329)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     579..598
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             919..939
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             952..996
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     3'UTR           1041..1166
                     /label=Xenopus globin 3'-UTR
                     /note="3'-UTR of the major beta-globin gene of Xenopus 
                     laevis"
     primer_bind     complement(1265..1281)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1265..1281)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1278..1300)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1289..1305
                     /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(1313..1319,1320..1337,1338..1343))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1358..1379
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1496..1513)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1667..2255)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1747..1766)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2426..3217,3218..3286))
                     /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     3049..3068
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3287..3391)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3459..3477
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3515..3537)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3637..3656
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3870..3889
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 atttaggtga cactatagaa tacaagcttg cttgttcttt ttgcagaagc tcagaataaa
       61 cgctcaactt tggcagatct gatcatgcca tgggggccca tgctgtgctg tatgagaaga
      121 accaaacagg ttgaaaagaa tgatgacgac caaaagattg gcggcatggt gagcaagggc
      181 gaggaggata acatggccat catcaaggag ttcatgcgct tcaaggtgca catggagggc
      241 tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc
      301 acccagaccg ccaagctgaa ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc
      361 ctgtcccctc agttcatgta cggctccaag gcctacgtga agcaccccgc cgacatcccc
      421 gactacttga agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag
      481 gacggcggcg tggtgaccgt gacccaggac tcctccctgc aggacggcga gttcatctac
      541 aaggtgaagc tgcgcggcac caacttcccc tccgacggcc ccgtaatgca gaagaagacc
      601 atgggctggg aggcctcctc cgagcggatg taccccgagg acggcgccct gaagggcgag
      661 atcaagcaga ggctgaagct gaaggacggc ggccactacg acgctgaggt caagaccacc
      721 tacaaggcca agaagcccgt gcagctgccc ggcgcctaca acgtcaacat caagttggac
      781 atcacctccc acaacgagga ctacaccatc gtggaacagt acgaacgcgc cgagggccgc
      841 cactccaccg gcggcatgga cgagctgtac aagtccggag gggatcggtg gagctccacc
      901 ggtggcggcc gctctagaga gaatctttat tttcagggcg ccgccaaatt caaagaaacc
      961 gctgctgcta aattcgaacg ccagcacatg gacagcggag gtggaggtac tagtgactga
     1021 ctaggatctg gttaccacta aaccagcctc aagaacaccc gaatggagtc tctaagctac
     1081 ataataccaa cttacacttt acaaaatgtt gtcccccaaa atgtagccat tcgtatctgc
     1141 tcctaataaa aaraaagttt cttcacattc taaaaaaaaa aaaaaaaaaa aaaaaaammc
     1201 cccccccccc ccccctgcag gtcgactcta gaggatcccc gggcgagctc gaattcgtaa
     1261 tcatgtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     1321 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     1381 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     1441 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     1501 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     1561 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     1621 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     1681 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     1741 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     1801 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     1861 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     1921 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     1981 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     2041 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     2101 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     2161 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     2221 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     2281 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     2341 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     2401 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     2461 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     2521 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     2581 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     2641 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     2701 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     2761 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     2821 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     2881 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     2941 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     3001 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     3061 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     3121 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     3181 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     3241 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     3301 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     3361 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     3421 tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct
     3481 ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga
     3541 cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag
     3601 cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga
     3661 gagtgcacca ttcgacgctc tcccttatgc gactcctgca ttaggaagca gcccagtagt
     3721 aggttgaggc cgttgagcac cgccgccgca aggaatggtg catgcaagga gatggcgccc
     3781 aacagtcccc cggccacggg gcctgccacc atacccacgc cgaaacaagc gctcatgagc
     3841 ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg cgatataggc gccagcaacc
     3901 gcacctgtgg cgccggtgat gccggccacg atgcgtccgg cgtagaggat ctggctagcg
     3961 atgaccctgc tgattggttc gctgaccatt tccgggtgcg ggacggcgtt accagaaact
     4021 cagaaggttc gtccaaccaa accgactctg acggcagttt acgagagaga tgatagggtc
     4081 tgcttcagta agccagatgc tacacaatta ggcttgtaca tattgtcgtt agaacgcggc
     4141 tacaattaat acataacctt atgtatcata cacatacg
//
