LOCUS       sequence-429609-        3508 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Aspergillus nidulans codon-adjusted mStayGold fluorescent protein, 
            includes linker for C-terminal tagging..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3508)
  AUTHORS   Modaffari D, Finlayson A, Miao Y, Wallace EWJ, Sawin KE
  TITLE     Improved gene editing and fluorescent-protein tagging in Aspergillus
            nidulans using a Golden Gate-based CRISPR-Cas9 plasmid system.
  JOURNAL   Wellcome Open Res. 2024 Oct 17;9:602. doi: 
            10.12688/wellcomeopenres.23086.1. eCollection 2024.
  PUBMED    39640368
REFERENCE   2  (bases 1 to 3508)
  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: "Wellcome 
            Open Res."; date: "2024-10-17"; volume: "9:602. doi"; pages: " 
            10.12688/wellcomeopenres.23086.1. eCollection 2024"
COMMENT     Sequence Label: pDM098
FEATURES             Location/Qualifiers
     source          1..3508
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     378..395
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             420..443
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             561..1238
                     /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(1287..1303)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1287..1303)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1300..1322)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1311..1327
                     /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(1335..1341,1342..1359,1360..1365))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1380..1401
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1518..1535)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1689..2277)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1769..1788)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2448..3239,3240..3308))
                     /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     3071..3090
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3309..3413)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3481..3499
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt cgagctcggt acccggggag
      421 actacaagga cgacgacgac aaatccggtg gttctagcgg tggttcttca ggttccgaga
      481 ctcctggtac ctccgaatct gccacccctg aatcaggcgg aggtggatca ggtggtggag
      541 gatctggagg tggtggttct atggtcagca ccggtgagga gctcttcacc ggtgtcgtcc
      601 ctttcaagtt ccagctcaag ggtaccatca acggtaagtc cttcaccgtc gagggtgagg
      661 gtgaaggtaa ctcccatgaa ggttcccaca agggtaagta cgtctgtacc tctggtaaac
      721 tccctatgtc ctgggccgct ctcggtacct cctttggcta tggtatgaag tactacacca
      781 agtacccctc cggtctgaag aactggttcc acgaggtcat gcctgaaggt ttcacctacg
      841 accgccacat ccagtacaag ggtgacggtt ccatccacgc caagcaccag cacttcatga
      901 agaacggtac ctaccacaac atcgtcgagt tcaccggtca ggacttcaag gagaactccc
      961 ccgttctcac cggtgatatg gacgtttccc tccccaacga agtccagcac atccccatcg
     1021 acgatggtgt tgagtgcacc gtcaccctcc agtatcccct cctctccgat gagtctaagt
     1081 gcgtcgaggc ctaccagaac accatcatca agcccctcca caaccaacct gcccctgatg
     1141 tccccttcca ttggatccgc aagcagtaca cccagtccaa ggacgatacc gaagagcgcg
     1201 accacatcat ccagtccgag actctcgaag ctcaccttta atcctctaga gtcgacctgc
     1261 aggcatgcaa gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     1321 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     1381 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     1441 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     1501 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     1561 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     1621 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     1681 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     1741 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     1801 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     1861 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     1921 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     1981 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     2041 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     2101 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     2161 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     2221 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     2281 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     2341 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     2401 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     2461 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     2521 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     2581 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     2641 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     2701 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     2761 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     2821 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     2881 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     2941 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     3001 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     3061 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     3121 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     3181 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     3241 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     3301 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     3361 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     3421 ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa
     3481 aataggcgta tcacgaggcc ctttcgtc
//
