LOCUS       sequence-430130-        3442 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Aspergillus nidulans codon-adjusted mStayGold2 fluorescent protein, 
            includes linker for N-terminal tagging..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3442)
  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 3442)
  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: pDM069
FEATURES             Location/Qualifiers
     source          1..3442
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(475..1266,1267..1335))
                     /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     1098..1117
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1336..1440)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1508..1526
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1564..1586)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1686..1705
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1899..1921
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1913..1930
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1914..1930
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             1988..2665
                     /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(2756..2772)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2756..2772)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2769..2791)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2780..2796
                     /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(2804..2810,2811..2828,2829..2834))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2849..2870
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2987..3004)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(3158..3442,1..304))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3238..3257)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
       61 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
      121 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
      181 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
      241 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
      301 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
      361 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
      421 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
      481 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
      541 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
      601 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
      661 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
      721 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
      781 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
      841 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
      901 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
      961 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     1021 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     1081 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     1141 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     1201 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     1261 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     1321 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     1381 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     1441 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
     1501 ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga
     1561 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
     1621 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa
     1681 ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca
     1741 cagatgcgta aggagaaaat accgcatcag gcgccattcg ccattcaggc tgcgcaactg
     1801 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg
     1861 tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac
     1921 gacggccagt gaattcgagc tcggtacccg gggatcccaa gctgctgctt tagaattagt
     1981 tgatcctatg gtctccaccg gtgaggagct cttcaccggt gtcgtcccct tcaagttcca
     2041 gctcaagggt accatcaacg gtaagtcctt caccgtcgag ggtgagggtg agggtaactc
     2101 ccacgagggt tcccacaagg gtaagtacgt ctgcacctcc ggtaagctcc ccatgtcctg
     2161 ggccgccctc ggtacctcct tcggttacgg tatgaagtac tacaccaagt acccctccgg
     2221 tctcaagaac tggttccacg aggtcatgcc cgagggtttc acctacgacc gccacatcca
     2281 gtacaagggt gacggttcca tccacgccaa gcaccagcac ttcatgaaga acggtaccta
     2341 ccacaacatc gtcgagttca ccggtcagga cttcaaggag aactcccccg tcctcaccgg
     2401 tgacatggac gtctccctcc ccaacgaggt ccagcacatc cccatcgacg acggtgtcga
     2461 gtgcaccgtc accctccagt accccctcct ctccgacgag tccaagtgcg tcgaggccta
     2521 ccagaacacc atcatcaagc ccctccacaa ccagcccgcc cccgacgtcc ccttccactg
     2581 gatccgcaag cagtacaccc agtccaagga cgacaccgag gagcgcgacc acatcatcca
     2641 gtccgagacc ctcgaggccc acctctactc ccgcaccaag ctcgagaaca tcctcggtcc
     2701 catccgcgga tcctctagag tcgacctgca ggcatgcaag cttggcgtaa tcatggtcat
     2761 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     2821 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     2881 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     2941 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     3001 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     3061 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     3121 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     3181 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     3241 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     3301 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     3361 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     3421 cccccgttca gcccgaccgc tg
//
