LOCUS       sequence-430145-        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     161..179
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(217..239)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     339..358
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     552..574
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     566..583
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     567..583
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1412..1428)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1412..1428)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1425..1447)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1436..1452
                     /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(1460..1466,1467..1484,1485..1490))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1505..1526
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1643..1660)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1814..2402)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1894..1913)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2573..3364,3365..3433))
                     /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     3196..3215
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(3434..3445,1..93))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
       61 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
      121 cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc
      181 ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
      241 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
      301 agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact
      361 gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat
      421 caggcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
      481 ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac
      541 gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgaattcg agctcggtac
      601 ccggggatcc atggtctcca agggtgagga gctcatcaag gagaacatgc acatgaagct
      661 ctacatggag ggtaccgtca acaaccacca cttcaagtgc accaccgaag gtgagggtaa
      721 gccctacgag ggtacccaga cccagcgcat caaagttgtc gaaggtggtc ccctcccttt
      781 cgctttcgac atcctcgcca cctgcttcat gtacggttcc aagaccttca tcaaccacac
      841 ccagggtatc cccgacttct tcaagcagtc cttccccgag ggtttcacct gggagcgcgt
      901 tactacctac gaggatggtg gtgtcctcac tgtcacccaa gacacctccc ttcaggacgg
      961 ttgcctcatc tacaacgtca agctccgcgg tgtcaacttc ccctccaacg gtcccgtcat
     1021 gcagaagaag acccttggtt gggaagctac tactgagacc ctttaccccg ccgacggtgg
     1081 tctcgagggt cgttgtgaca tggccttaaa gctcgttggt ggtggtcact tacactgcaa
     1141 cctcaagacc acctaccgct ccaagaagcc cgccaagaac ctcaagatgc ccggtgtcta
     1201 ctttgttgac cgtcgccttg agcgcatcaa ggaggccgac aacgagacct acgttgaaca
     1261 acacgaggtc gccgttgctc gctactgcga cctcccttct aagctcggtc acaagctcaa
     1321 cggtatggac gagctctaca agaacatcct cggtcccatc cgcggatcct ctagagtcga
     1381 cctgcaggca tgcaagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     1441 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     1501 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     1561 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     1621 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     1681 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     1741 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     1801 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     1861 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     1921 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     1981 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     2041 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     2101 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     2161 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     2221 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     2281 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     2341 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     2401 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     2461 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     2521 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     2581 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     2641 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     2701 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     2761 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     2821 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     2881 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     2941 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     3001 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     3061 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     3121 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     3181 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     3241 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     3301 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     3361 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     3421 gttgaatact catactcttc ctttt
//
