LOCUS       sequence-374654-        5073 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  for CALI .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5073)
  AUTHORS   Wavreil FDM, Poon J, Wessel GM, Yajima M
  TITLE     Light-induced, spatiotemporal control of protein in the developing 
            embryo of the sea urchin.
  JOURNAL   Dev Biol. 2021 Oct;478:13-24. doi: 10.1016/j.ydbio.2021.06.006. Epub
            2021 Jun 18.
  PUBMED    34147471
REFERENCE   2  (bases 1 to 5073)
  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"; journalName: "Dev Biol.";
            date: "2021-10"; volume: "478:13-24. doi"; pages: " 
            10.1016/j.ydbio.2021.06.006. Epub 2021 Jun 18"
COMMENT     Sequence Label: miniSOG-GFP
FEATURES             Location/Qualifiers
     source          1..5073
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     5'UTR           14..56
                     /label=Xenopus globin 5'-UTR
                     /note="translational enhancer from the 5'-UTR of the major 
                     beta-globin gene of Xenopus laevis"
     CDS             79..396
                     /codon_start=1
                     /product="mini singlet oxygen generator, a fluorescent 
                     flavoprotein suitable for correlated light and electron 
                     microscopy (Shu et al., 2011)"
                     /label=miniSOG
                     /translation="MEKSFVITDPRLPDNPIIFASDGFLELTEYSREEILGRNGRFLQG
                     PETDQATVQKIRDAIRDQREITVQLINYTKSGKKFWNLLHLQPMRDQKGELQYFIGVQL
                     DG"
     CDS             join(403..405,406..408,409..1119)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(403..405,406..408,409..1119)
                     /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(448..469)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(709..728)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1056..1077
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1797..1817
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             1830..1874
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     3'UTR           1919..2044
                     /label=Xenopus globin 3'-UTR
                     /note="3'-UTR of the major beta-globin gene of Xenopus 
                     laevis"
     primer_bind     complement(2143..2159)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2143..2159)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2156..2178)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2167..2183
                     /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(2191..2197,2198..2215,2216..2221))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2236..2257
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2374..2391)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2545..3133)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2625..2644)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3304..4095,4096..4164))
                     /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     3927..3946
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4165..4269)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4337..4355
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4393..4415)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4515..4534
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4748..4767
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        join(5057..5073,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(5057..5073,1)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gaatacaagc ttgcttgttc tttttgcaga agctcagaat aaacgctcaa ctttggcaga
       61 tctatgccat cggggcccat ggagaaaagt ttcgtgataa ctgatccacg gctgccagac
      121 aatcccatca tcttcgcatc cgatggcttc ctggagctga ccgagtattc cagagaggag
      181 atcctgggcc gcaatggccg ctttctgcag ggaccagaga cagaccaggc cacagtgcag
      241 aagattcgcg atgccattag agatcagcgc gagattaccg tgcagctgat aaactacaca
      301 aaaagcggga agaaattctg gaacctcctg cacctccagc ccatgaggga ccagaagggt
      361 gagctccagt atttcatcgg agtgcagctg gatggagggc ccatggtgag caagggcgag
      421 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
      481 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
      541 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc
      601 tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag
      661 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac
      721 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
      781 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
      841 aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc
      901 aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
      961 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc
     1021 gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
     1081 gccgccggga tcactctcgg catggacgag ctatacaagt ccggagggga tcggtggagc
     1141 tccaccggtg gcggccgcat gccagaactt ggatttagca tatcaggtgg tgtcgggggt
     1201 agaggaaacc cattcagacc tgatgatgat ggtatatttg taacaagggt acaacctgaa
     1261 ggaccagcat caaaattact gcagccaggt gataaaatta ttcaggctaa tggctacagt
     1321 tttataaata ttgaacatgg acaagcagtg tccttgctaa aaactttcca gaatacagtt
     1381 gaactcatca ttgtacgaga agttggtaac ggtgcaaaac aagagattcg agtgagggtt
     1441 gaaaaagacg gtggttctgg aggcgtttct tctgttccga ccaatctgga agttgttgct
     1501 gcgaccccga ctagcctgct gatcagctgg gatgcgtata gggagcttcc ggttagttat
     1561 taccgtatca cgtacggtga aaccggtggt aactccccgg ttcaggaatt cactgtacct
     1621 ggttccaagt ctactgctac catcagcggc ctgaaaccgg gtgtcgacta taccatcact
     1681 gtatacgcac attacaacta ccattactac tctagcccaa tctcgattaa ctaccgtacc
     1741 tctagactcg agctcaagct tcgaattctg cagtcgacgg tagcggccgc tctagagaga
     1801 atctttattt tcagggcgcc gccaaattca aagaaaccgc tgctgctaaa ttcgaacgcc
     1861 agcacatgga cagcggaggt ggaggtacta gtgactgact aggatctggt taccactaaa
     1921 ccagcctcaa gaacacccga atggagtctc taagctacat aataccaact tacactttac
     1981 aaaatgttgt cccccaaaat gtagccattc gtatctgctc ctaataaaaa gaaagtttct
     2041 tcacattcta aaaaaaaaaa aaaaaaaaaa aaaaaacccc cccccccccc ccctgcaggt
     2101 cgactctaga ggatccccgg gcgagctcga attcgtaatc atgtcatagc tgtttcctgt
     2161 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     2221 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     2281 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     2341 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     2401 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     2461 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     2521 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     2581 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     2641 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     2701 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     2761 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     2821 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     2881 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     2941 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     3001 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     3061 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     3121 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     3181 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     3241 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     3301 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     3361 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     3421 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     3481 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     3541 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     3601 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     3661 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     3721 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     3781 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     3841 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     3901 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     3961 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     4021 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     4081 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     4141 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     4201 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     4261 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat
     4321 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga
     4381 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
     4441 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
     4501 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccatt cgacgctctc
     4561 ccttatgcga ctcctgcatt aggaagcagc ccagtagtag gttgaggccg ttgagcaccg
     4621 ccgccgcaag gaatggtgca tgcaaggaga tggcgcccaa cagtcccccg gccacggggc
     4681 ctgccaccat acccacgccg aaacaagcgc tcatgagccc gaagtggcga gcccgatctt
     4741 ccccatcggt gatgtcggcg atataggcgc cagcaaccgc acctgtggcg ccggtgatgc
     4801 cggccacgat gcgtccggcg tagaggatct ggctagcgat gaccctgctg attggttcgc
     4861 tgaccatttc cgggtgcggg acggcgttac cagaaactca gaaggttcgt ccaaccaaac
     4921 cgactctgac ggcagtttac gagagagatg atagggtctg cttcagtaag ccagatgcta
     4981 cacaattagg cttgtacata ttgtcgttag aacgcggcta caattaatac ataaccttat
     5041 gtatcataca catacgattt aggtgacact ata
//
