LOCUS       sequence-430144-        3676 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Aspergillus nidulans codon-adjusted mStayGold fluorescent protein, 
            includes linker for N-terminal or internal tagging..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3676)
  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 3676)
  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: pDM097
FEATURES             Location/Qualifiers
     source          1..3676
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        17..121
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(122..190,191..982)
                     /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(340..359)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1153..1741
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1642..1661
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1895..1912
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2029..2050
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2065..2070,2071..2088,2089..2095)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2103..2119
                     /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)."
     primer_bind     2108..2130
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2127..2143
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2127..2143
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             complement(2189..2212)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(2360..3037)
                     /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"
     CDS             complement(3155..3178)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(3203..3220)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3203..3219)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3212..3234)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(3428..3447)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3547..3569
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3607..3625)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
       61 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
      121 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
      181 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
      241 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
      301 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
      361 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
      421 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
      481 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
      541 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
      601 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
      661 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
      721 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
      781 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
      841 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
      901 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
      961 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     1021 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     1081 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     1141 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     1201 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     1261 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     1321 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     1381 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     1441 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     1501 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     1561 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     1621 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     1681 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     1741 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     1801 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     1861 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     1921 agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg
     1981 gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta
     2041 gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg
     2101 aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagct
     2161 tgcatgcctg caggtcgact ctagaggact tgtcatcgtc atccttataa tcggaaccgc
     2221 cagaagaacc accagaggac tcaggagtag cggactcaga ggtaccaggg gtttcggaac
     2281 cggatgaacc acctccacca gatccgccac caccagaacc accaccacca acagcagaag
     2341 cagaaggttc gtgccaggga aggtgagctt cgagagtctc ggactggatg atgtggtcgc
     2401 gctcttcggt atcgtccttg gactgggtgt actgcttgcg gatccaatgg aaggggacat
     2461 caggggcagg ttggttgtgg aggggcttga tgatggtgtt ctggtaggcc tcgacgcact
     2521 tagactcatc ggagaggagg ggatactgga gggtgacggt gcactcaaca ccatcgtcga
     2581 tggggatgtg ctggacttcg ttggggaggg aaacgtccat atcaccggtg agaacggggg
     2641 agttctcctt gaagtcctga ccggtgaact cgacgatgtt gtggtaggta ccgttcttca
     2701 tgaagtgctg gtgcttggcg tggatggaac cgtcaccctt gtactggatg tggcggtcgt
     2761 aggtgaaacc ttcaggcatg acctcgtgga accagttctt cagaccggag gggtacttgg
     2821 tgtagtactt cataccatag ccaaaggagg taccgagagc ggcccaggac atagggagtt
     2881 taccagaggt acagacgtac ttacccttgt gggaaccttc atgggagtta ccttcaccct
     2941 caccctcgac ggtgaaggac ttaccgttga tggtaccctt gagctggaac ttgaaaggga
     3001 cgacaccggt gaagagctcc tcaccggtgc tgaccataga accaccacct ccagatcctc
     3061 caccacctga tccacctccg cctgattcag gggtggcaga ttcggaggta ccaggagtct
     3121 cggaacctga agaaccaccg ctagaaccac cggatttgtc gtcgtcgtcc ttgtagtctc
     3181 cccgggtacc gagctcgaat tcactggccg tcgttttaca acgtcgtgac tgggaaaacc
     3241 ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata
     3301 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggc
     3361 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atatggtgca
     3421 ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac
     3481 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga
     3541 ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac
     3601 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     3661 agacgtcagg tggcac
//
