LOCUS       sequence-419228-        3566 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mStayGold has an n1 adaptor at its N-terminus and is useful for 
            C-terminal tagging. This construct is used also for expression of 
            mStayGold itself..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3566)
  AUTHORS   Ando R, Shimozono S, Ago H, Takagi M, Sugiyama M, Kurokawa H, Hirano
            M, Niino Y, Ueno G, Ishidate F, Fujiwara T, Okada Y, Yamamoto M, 
            Miyawaki A
  TITLE     StayGold variants for molecular fusion and membrane-targeting 
            applications.
  JOURNAL   Nat Methods. 2023 Nov 30. doi: 10.1038/s41592-023-02085-6.
  PUBMED    38036853
REFERENCE   2  (bases 1 to 3566)
  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: "Nat 
            Methods. 2023 Nov 30. doi: 10.1038/s41592-023-02085-6."
COMMENT     Sequence Label: pRSETB/mStayGold
FEATURES             Location/Qualifiers
     source          1..3566
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     20..39
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        20..38
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             70..92
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             112..129
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             133..165
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             join(169..177,178..192)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             199..876
                     /codon_start=1
                     /product="bright and highly photostable monomeric green 
                     fluorescent protein (Ando et al., 2023)"
                     /label=mStayGold(J)
                     /note="includes multiple mutations plus an N-terminal 
                     iinker sequence"
                     /translation="MVSTGEELFTGVVPFKFQLKGTINGKSFTVEGEGEGNSHEGSHKG
                     KYVCTSGKLPMSWAALGTSFGYGMKYYTKYPSGLKNWFHEVMPEGFTYDRHIQYKGDGS
                     IHAKHQHFMKNGTYHNIVEFTGQDFKENSPVLTGDMDVSLPNEVQHIPIDDGVECTVTL
                     QYPLLSDESKCVEAYQNTIIKPLHNQPAPDVPFHWIRKQYTQSKDDTEERDHIIQSETL
                     EAHL"
     primer_bind     complement(967..985)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      981..1028
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1125..1580
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1212..1231)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1422..1443
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1606..1710
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1711..1779,1780..2571)
                     /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     complement(1929..1948)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2742..3330
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3231..3250
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3484..3501
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gatctcgatc ccgcgaaatt aatacgactc actataggga gaccacaacg gtttccctct
       61 agaaataatt ttgtttaact ttaagaagga gatatacata tgcggggttc tcatcatcat
      121 catcatcatg gtatggctag catgactggt ggacagcaaa tgggtcggga tctgtacgac
      181 gatgacgata aggatccgat ggtgtctaca ggcgaggagc tgtttaccgg cgtggtgccc
      241 ttcaagttcc agctgaaggg caccatcaac ggcaagagct tcaccgtgga aggcgagggc
      301 gagggcaata gccacgaggg cagccacaaa ggcaagtacg tgtgcaccag cggcaaactg
      361 ccaatgtctt gggccgccct gggaactagc ttcggctatg gcatgaagta ctacaccaag
      421 taccccagcg gcctgaagaa ctggttccac gaggtgatgc ccgagggctt cacctacgac
      481 agacacatcc agtacaaggg cgacggcagc atccacgcca agcaccagca cttcatgaag
      541 aacggcacct accacaacat cgtggagttc accggccagg acttcaagga gaacagcccc
      601 gtgctgaccg gcgacatgga cgtgagcctg cccaacgagg tgcagcacat ccccatagat
      661 gacggcgtgg agtgcacagt gaccctgcag taccctctgc tgagcgacga gagcaagtgc
      721 gtggaggcct accagaacac catcatcaag cccctgcaca atcagccagc ccccgatgtg
      781 ccattccact ggatcagaaa gcagtacacc cagagcaagg acgacaccga ggagagagac
      841 cacatcatcc agagcgagac cctggaggcc cacctgtaac tcgagatctg cagctggtac
      901 catggaattc gaagcttgat ccggctgcta acaaagcccg aaaggaagct gagttggctg
      961 ctgccaccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg gtcttgaggg
     1021 gttttttgct gaaaggagga actatatccg gatctggcgt aatagcgaag aggcccgcac
     1081 cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc cctgtagcgg
     1141 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     1201 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     1261 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     1321 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     1381 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     1441 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     1501 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     1561 aatattaacg cttacaattt aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     1621 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     1681 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     1741 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     1801 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     1861 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     1921 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     1981 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     2041 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     2101 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     2161 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     2221 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     2281 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     2341 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     2401 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     2461 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     2521 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     2581 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     2641 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     2701 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     2761 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     2821 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     2881 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     2941 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     3001 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     3061 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     3121 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     3181 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     3241 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     3301 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     3361 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     3421 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     3481 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     3541 gcgcgttggc cgattcatta atgcag
//
