LOCUS       sequence-430143-        3418 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Aspergillus nidulans codon-adjusted mStayGold2 fluorescent protein, 
            includes linker for C-terminal tagging..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3418)
  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 3418)
  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: pDM067
FEATURES             Location/Qualifiers
     source          1..3418
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     124..143
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(362..466)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     534..552
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(590..612)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     712..731
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     925..947
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     939..956
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     940..956
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             1011..1688
                     /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(1758..1774)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1758..1774)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1771..1793)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1782..1798
                     /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(1806..1812,1813..1830,1831..1836))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1851..1872
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1989..2006)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2160..2748)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2240..2259)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2919..3418,1..292,293..361))
                     /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"
ORIGIN
        1 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
       61 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
      121 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
      181 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
      241 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
      301 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
      361 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
      421 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
      481 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
      541 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
      601 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
      661 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag
      721 agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga
      781 gaaaataccg catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat
      841 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat
      901 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgaat
      961 tcgagctcgg tacccgggga caagctgctg ctttagaatt agttgatcct atggtctcca
     1021 ccggtgagga gctcttcacc ggtgtcgtcc ccttcaagtt ccagctcaag ggtaccatca
     1081 acggtaagtc cttcaccgtc gagggtgagg gtgagggtaa ctcccacgag ggttcccaca
     1141 agggtaagta cgtctgcacc tccggtaagc tccccatgtc ctgggccgcc ctcggtacct
     1201 ccttcggtta cggtatgaag tactacacca agtacccctc cggtctcaag aactggttcc
     1261 acgaggtcat gcccgagggt ttcacctacg accgccacat ccagtacaag ggtgacggtt
     1321 ccatccacgc caagcaccag cacttcatga agaacggtac ctaccacaac atcgtcgagt
     1381 tcaccggtca ggacttcaag gagaactccc ccgtcctcac cggtgacatg gacgtctccc
     1441 tccccaacga ggtccagcac atccccatcg acgacggtgt cgagtgcacc gtcaccctcc
     1501 agtaccccct cctctccgac gagtccaagt gcgtcgaggc ctaccagaac accatcatca
     1561 agcccctcca caaccagccc gcccccgacg tccccttcca ctggatccgc aagcagtaca
     1621 cccagtccaa ggacgacacc gaggagcgcg accacatcat ccagtccgag accctcgagg
     1681 cccacctcta ctcccgcacc aagctcgagt aatcctctag agtcgacctg caggcatgca
     1741 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     1801 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     1861 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     1921 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     1981 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     2041 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     2101 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     2161 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     2221 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     2281 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     2341 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     2401 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     2461 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     2521 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     2581 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     2641 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     2701 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     2761 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     2821 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     2881 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     2941 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     3001 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     3061 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     3121 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     3181 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     3241 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     3301 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     3361 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgc
//
