LOCUS       sequence-430146-        3403 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Aspergillus nidulans codon-adjusted mScarlet3 fluorescent protein, 
            includes linker for N-terminal tagging..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3403)
  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 3403)
  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: pDM043
FEATURES             Location/Qualifiers
     source          1..3403
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     82..99
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    216..237
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(252..257,258..275,276..282)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    290..306
                     /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     295..317
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     314..330
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     314..330
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             complement(408..1094)
                     /codon_start=1
                     /product="bright cysteine-free monomeric red fluorescent 
                     protein with fast and complete maturation (Gadella et al., 
                     2023)"
                     /label=mScarlet3
                     /translation="MDSTEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTAK
                     LRVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYWKQSFPEGFKWERVMNFEDGGA
                     VSVAQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDVVLKGDIKM
                     ALRLKDGGRYLADFKTTYRAKKPVQMPGAFNIDRKLDITSHNEDYTVVEQYERSVARHS
                     TGGSGGS"
     primer_bind     complement(1117..1134)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1117..1133)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1126..1148)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1342..1361)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1461..1483
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1521..1539)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1607..1711
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1712..1780,1781..2572)
                     /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(1930..1949)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2743..3331
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3232..3251
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
       61 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
      121 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
      181 cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag
      241 gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga
      301 taacaatttc acacaggaaa cagctatgac catgattacg ccaagcttgc atgcctgcag
      361 gtcgactcta gaggatcctg agcagcagcg agctcgagga cgtcagggga accaccagaa
      421 ccaccggtag aatgacgagc aacggaacgc tcatattgtt cgacgacggt gtagtcctcg
      481 ttgtgggagg tgatgtcgag cttgcggtcg atgttgaagg caccaggcat ctgaacgggc
      541 tttttagcac ggtaggtggt cttgaagtca gcgaggtagc gaccaccgtc cttaaggcga
      601 agggccatct tgatgtcacc tttaagaacg acatcctcag ggtagagacg ttcggtggag
      661 gcctcccaac ccatggtctt cttctgcatg acgggaccat caggaggaaa attggtaccg
      721 cggagcttga ccttgtagat gagggtaccg tcctcaaggg aagtatcttg ggcaacagag
      781 acggcaccac catcttcgaa gttcatgacg cgctcccact tgaaaccctc gggaaaggac
      841 tgcttccaat aatcgggaat atcagcgggg tgcttggtga aggcgcggga accgtacatg
      901 aactgaggag agaggatgtc ccaggagaaa ggtaaaggac cacctttagt aacacgtaac
      961 ttggcggtct gggtaccctc gtaggggcga ccctcaccct caccctcgat ctcgaactcg
     1021 tgaccgttca tagaaccttc catgtgaacc ttaaaacgca taaactcctt gatgacggcc
     1081 tcggtggagt ccatcccggg taccgagctc gaattcactg gccgtcgttt tacaacgtcg
     1141 tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc
     1201 cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct
     1261 gaatggcgaa tggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     1321 ccgcatatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg
     1381 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
     1441 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
     1501 gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat
     1561 aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     1621 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     1681 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     1741 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     1801 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     1861 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     1921 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     1981 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     2041 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     2101 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     2161 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     2221 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     2281 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     2341 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     2401 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     2461 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     2521 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     2581 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     2641 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     2701 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     2761 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     2821 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     2881 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     2941 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     3001 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     3061 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     3121 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     3181 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     3241 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     3301 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     3361 cctggccttt tgctggcctt ttgctcacat gttctttcct gcg
//
