LOCUS       sequence-261755-        2720 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of EMTB-GFP-ePDZ mRNA for microinjection for use of 
            optogenetics in S. purpuratus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2720)
  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 2720)
  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-EMTB-GFP-ePDZ
FEATURES             Location/Qualifiers
     source          1..2720
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     5'UTR           6..48
                     /label=Xenopus globin 5'-UTR
                     /note="translational enhancer from the 5'-UTR of the major 
                     beta-globin gene of Xenopus laevis"
     CDS             61..90
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             join(922..924,925..927,928..1638)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(922..924,925..927,928..1638)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(967..988)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1228..1247)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1575..1596
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2316..2336
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             2349..2393
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     3'UTR           2438..2563
                     /label=Xenopus globin 3'-UTR
                     /note="3'-UTR of the major beta-globin gene of Xenopus 
                     laevis"
     primer_bind     complement(2662..2678)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2662..2678)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2675..2697)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2686..2702
                     /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)."
ORIGIN
        1 gcttgcttgt tctttttgca gaagctcaga ataaacgctc aactttggca gatctgaatg
       61 gagcagaagc tcatctcaga agaagacctc ggcgcgccag tgcgaagcga aacagcaccc
      121 gacagctaca aagtgcaaga taagaaaaat gcctccagcc gccctgcctc tgcaatttca
      181 ggacaaaata acaaccactc aggaaataaa ccagaccctc cgcctgtgtt acgtgttgat
      241 gaccggcagc ggctggcccg ggagcgacgt gaggaacggg agaaacagct agctgcaaga
      301 gaaatagtgt ggttagaaag agaagagcga gccaggcagc actacgagaa gcacctggaa
      361 gagcggaaga agaggttgga ggagcagagg cagaaggagg agcggaggag ggctgctgtg
      421 gaggagaagc ggaggcagag acttgaggag gacaaagaac gccacgaagc tgttgtacgg
      481 cgcacaatgg aaaggagcca gaagccaaaa cagaagcata accgttggtc gtggggaggc
      541 tctctccatg ggagccctag catccacagt gcagatccag acaggcggtc agtttccacc
      601 atgaatcttt cgaaatatgt tgatcccgtc attagcaagc ggctctcctc ttcatctgca
      661 actttactaa attctccaga tagagctcgc cgcctgcagc tcagcccatg ggagagcagc
      721 gttgttaaca gactcctgac gcccacacat tcgttcctgg ccagaagtaa aagcacagct
      781 gccttgtctg gagaagcagc atcttgcagc cccatcatca tgccctacaa agctgcacac
      841 tctagaaatt cgatggatcg accaaaactc tttgtaacac cacctgaggg ctcttcggac
      901 ccaccggtcg ccaccgggcc catggtgagc aagggcgagg agctgttcac cggggtggtg
      961 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
     1021 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
     1081 ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc
     1141 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     1201 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     1261 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     1321 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
     1381 atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag
     1441 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     1501 gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac
     1561 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     1621 atggacgagc tatacaagtc cggaggggat cggtggagct ccaccggtgg cggccgcatg
     1681 ccagaacttg gatttagcat atcaggtggt gtcgggggta gaggaaaccc attcagacct
     1741 gatgatgatg gtatatttgt aacaagggta caacctgaag gaccagcatc aaaattactg
     1801 cagccaggtg ataaaattat tcaggctaat ggctacagtt ttataaatat tgaacatgga
     1861 caagcagtgt ccttgctaaa aactttccag aatacagttg aactcatcat tgtacgagaa
     1921 gttggtaacg gtgcaaaaca agagattcga gtgagggttg aaaaagacgg tggttctgga
     1981 ggcgtttctt ctgttccgac caatctggaa gttgttgctg cgaccccgac tagcctgctg
     2041 atcagctggg atgcgtatag ggagcttccg gttagttatt accgtatcac gtacggtgaa
     2101 accggtggta actccccggt tcaggaattc actgtacctg gttccaagtc tactgctacc
     2161 atcagcggcc tgaaaccggg tgtcgactat accatcactg tatacgcaca ttacaactac
     2221 cattactact ctagcccaat ctcgattaac taccgtacct ctagactcga gctcaagctt
     2281 cgaattctgc agtcgacggt agcggccgct ctagagagaa tctttatttt cagggcgccg
     2341 ccaaattcaa agaaaccgct gctgctaaat tcgaacgcca gcacatggac agcggaggtg
     2401 gaggtactag tgactgacta ggatctggtt accactaaac cagcctcaag aacacccgaa
     2461 tggagtctct aagctacata ataccaactt acactttaca aaatgttgtc ccccaaaatg
     2521 tagccattcg tatctgctcc taataaaaag aaagtttctt cacattctaa aaaaaaaaaa
     2581 aaaaaaaaaa cccccccccc cccccccccc cctgcaggtc gactctagag gatccccggg
     2641 cgagctcgaa ttcgtaatca tgtcatagct gtttcctgtg tgaaattgtt atccgctcac
     2701 aattccacac aacatacgag
//
