LOCUS       sequence-422931-        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"
     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             453..1130
                     /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(1221..1237)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1221..1237)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1234..1256)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1245..1261
                     /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(1269..1275,1276..1293,1294..1299))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1314..1335
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1452..1469)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1623..2211)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1703..1722)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2382..3173,3174..3242))
                     /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     3005..3024
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3243..3347)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3415..3433
                     /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 acccggggat
      421 cccaagctgc tgctttagaa ttagttgatc ctatggtctc caccggtgag gagctcttca
      481 ccggtgtcgt ccccttcaag ttccagctca agggtaccat caacggtaag tccttcaccg
      541 tcgagggtga gggtgagggt aactcccacg agggttccca caagggtaag tacgtctgca
      601 cctccggtaa gctccccatg tcctgggccg ccctcggtac ctccttcggt tacggtatga
      661 agtactacac caagtacccc tccggtctca agaactggtt ccacgaggtc atgcccgagg
      721 gtttcaccta cgaccgccac atccagtaca agggtgacgg ttccatccac gccaagcacc
      781 agcacttcat gaagaacggt acctaccaca acatcgtcga gttcaccggt caggacttca
      841 aggagaactc ccccgtcctc accggtgaca tggacgtctc cctccccaac gaggtccagc
      901 acatccccat cgacgacggt gtcgagtgca ccgtcaccct ccagtacccc ctcctctccg
      961 acgagtccaa gtgcgtcgag gcctaccaga acaccatcat caagcccctc cacaaccagc
     1021 ccgcccccga cgtccccttc cactggatcc gcaagcagta cacccagtcc aaggacgaca
     1081 ccgaggagcg cgaccacatc atccagtccg agaccctcga ggcccacctc tactcccgca
     1141 ccaagctcga gaacatcctc ggtcccatcc gcggatcctc tagagtcgac ctgcaggcat
     1201 gcaagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     1261 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     1321 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     1381 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     1441 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     1501 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     1561 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     1621 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     1681 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     1741 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     1801 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     1861 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     1921 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     1981 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     2041 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     2101 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     2161 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     2221 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     2281 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     2341 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     2401 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     2461 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     2521 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     2581 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     2641 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     2701 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     2761 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     2821 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     2881 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     2941 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     3001 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     3061 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     3121 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     3181 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     3241 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     3301 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     3361 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
     3421 cgtatcacga ggccctttcg tc
//
