LOCUS       sequence-473510-        3571 bp    DNA     circular SYN 21-MAY-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 May 21, 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"
     rep_origin      727..1315
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1216..1235
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1469..1486
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     1571..1590
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1571..1589
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             1621..1643
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1663..1680
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1684..1716
                     /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(1720..1728,1729..1743)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             1750..2427
                     /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(2523..2541)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2537..2584
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2681..3136
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2768..2787)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2978..2999
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3162..3266
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3267..3335,3336..3571,1..556)
                     /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(3485..3504)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
       61 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
      121 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
      181 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
      241 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
      301 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
      361 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
      421 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
      481 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
      541 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
      601 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
      661 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
      721 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
      781 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
      841 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
      901 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
      961 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     1021 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     1081 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     1141 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     1201 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     1261 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     1321 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     1381 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     1441 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     1501 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca ggatctcgat
     1561 cccgcgaaat taatacgact cactataggg agaccacaac ggtttccctc tagaaataat
     1621 tttgtttaac tttaagaagg agatatacat atgcggggtt ctcatcatca tcatcatcat
     1681 ggtatggcta gcatgactgg tggacagcaa atgggtcggg atctgtacga cgatgacgat
     1741 aaggatccga tggtgtctac aggcgaggag ctgtttaccg gcgtggtgcc cttcaagttc
     1801 cagctgaagg gcaccatcaa cggcaagagc ttcaccgtgg aaggcgaggg cgagggcaat
     1861 agccacgagg gcagccacaa aggcaagtac gtgtgcacca gcggcaaact gccaatgtct
     1921 tgggccgccc tgggaactag cttcggctat ggcatgaagt actacaccaa gtaccccagc
     1981 ggcctgaaga actggttcca cgaggtgatg cccgagggct tcacctacga cagacacatc
     2041 cagtacaagg gcgacggcag catccacgcc aagcaccagc acttcatgaa gaacggcacc
     2101 taccacaaca tcgtggagtt caccggccag gacttcaagg agaacagccc cgtgctgacc
     2161 ggcgacatgg acgtgagcct gcccaacgag gtgcagcaca tccccataga tgacggcgtg
     2221 gagtgcacag tgaccctgca gtaccctctg ctgagcgacg agagcaagtg cgtggaggcc
     2281 taccagaaca ccatcatcaa gcccctgcac aatcagccag cccccgatgt gccattccac
     2341 tggatcagaa agcagtacac ccagagcaag gacgacaccg aggagagaga ccacatcatc
     2401 cagagcgaga ccctggaggc ccacctgcca tggcacgagc cttctgcttc tgccgtgtaa
     2461 gaattcgaag cttgatccgg ctgctaacaa agcccgaaag gaagctgagt tggctgctgc
     2521 caccgctgag caataactag cataacccct tggggcctct aaacgggtct tgaggggttt
     2581 tttgctgaaa ggaggaacta tatccggatc tggcgtaata gcgaagaggc ccgcaccgat
     2641 cgcccttccc aacagttgcg cagcctgaat ggcgaatggg acgcgccctg tagcggcgca
     2701 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     2761 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     2821 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     2881 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     2941 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     3001 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     3061 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     3121 ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     3181 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     3241 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     3301 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     3361 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     3421 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     3481 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     3541 gcatacacta ttctcagaat gacttggttg a
//
