LOCUS       sequence-422927-        3445 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Aspergillus nidulans codon-adjusted mCardinal fluorescent protein, 
            includes linker for N-terminal tagging..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3445)
  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 3445)
  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: pDM071
FEATURES             Location/Qualifiers
     source          1..3445
                     /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"
     primer_bind     complement(1224..1240)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1224..1240)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1237..1259)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1248..1264
                     /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(1272..1278,1279..1296,1297..1302))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1317..1338
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1455..1472)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1626..2214)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1706..1725)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2385..3176,3177..3245))
                     /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     3008..3027
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3246..3350)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3418..3436
                     /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 ccatggtctc caagggtgag gagctcatca aggagaacat gcacatgaag ctctacatgg
      481 agggtaccgt caacaaccac cacttcaagt gcaccaccga aggtgagggt aagccctacg
      541 agggtaccca gacccagcgc atcaaagttg tcgaaggtgg tcccctccct ttcgctttcg
      601 acatcctcgc cacctgcttc atgtacggtt ccaagacctt catcaaccac acccagggta
      661 tccccgactt cttcaagcag tccttccccg agggtttcac ctgggagcgc gttactacct
      721 acgaggatgg tggtgtcctc actgtcaccc aagacacctc ccttcaggac ggttgcctca
      781 tctacaacgt caagctccgc ggtgtcaact tcccctccaa cggtcccgtc atgcagaaga
      841 agacccttgg ttgggaagct actactgaga ccctttaccc cgccgacggt ggtctcgagg
      901 gtcgttgtga catggcctta aagctcgttg gtggtggtca cttacactgc aacctcaaga
      961 ccacctaccg ctccaagaag cccgccaaga acctcaagat gcccggtgtc tactttgttg
     1021 accgtcgcct tgagcgcatc aaggaggccg acaacgagac ctacgttgaa caacacgagg
     1081 tcgccgttgc tcgctactgc gacctccctt ctaagctcgg tcacaagctc aacggtatgg
     1141 acgagctcta caagaacatc ctcggtccca tccgcggatc ctctagagtc gacctgcagg
     1201 catgcaagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     1261 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     1321 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     1381 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     1441 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     1501 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     1561 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     1621 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     1681 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     1741 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     1801 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     1861 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     1921 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     1981 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     2041 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     2101 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     2161 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     2221 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     2281 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     2341 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     2401 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     2461 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     2521 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     2581 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     2641 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     2701 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     2761 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     2821 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     2881 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     2941 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     3001 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     3061 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     3121 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     3181 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     3241 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     3301 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     3361 cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat
     3421 aggcgtatca cgaggccctt tcgtc
//
