LOCUS       sequence-419226-        3571 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For N-terminal tagging with mStayGold. c4 is the appendage that 
            facilitates fusions to the C-terminus of mStayGold..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3571)
  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 3571)
  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(c4)
FEATURES             Location/Qualifiers
     source          1..3571
                     /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(972..990)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      986..1033
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1130..1585
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1217..1236)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1427..1448
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1611..1715
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1716..1784,1785..2576)
                     /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(1934..1953)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2747..3335
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3236..3255
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3489..3506
                     /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 cacctgccat ggcacgagcc ttctgcttct
      901 gccgtgtaag aattcgaagc ttgatccggc tgctaacaaa gcccgaaagg aagctgagtt
      961 ggctgctgcc accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt
     1021 gaggggtttt ttgctgaaag gaggaactat atccggatct ggcgtaatag cgaagaggcc
     1081 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggga cgcgccctgt
     1141 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     1201 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     1261 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     1321 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     1381 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     1441 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     1501 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     1561 aacaaaatat taacgcttac aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc
     1621 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     1681 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     1741 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     1801 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     1861 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     1921 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     1981 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     2041 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     2101 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     2161 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     2221 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     2281 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     2341 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     2401 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     2461 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     2521 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     2581 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     2641 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     2701 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     2761 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     2821 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     2881 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     2941 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     3001 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     3061 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     3121 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     3181 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     3241 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     3301 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     3361 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     3421 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     3481 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
     3541 tccccgcgcg ttggccgatt cattaatgca g
//
