LOCUS       sequence-429539-        3430 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Aspergillus nidulans codon-adjusted mNeonGreen fluorescent protein, 
            includes linker for C-terminal tagging..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3430)
  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 3430)
  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: pDM052
FEATURES             Location/Qualifiers
     source          1..3430
                     /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"
     regulatory      705..714
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(1209..1225)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1209..1225)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1222..1244)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1233..1249
                     /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(1257..1263,1264..1281,1282..1287))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1302..1323
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1440..1457)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1611..2199)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1691..1710)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2370..3161,3162..3230))
                     /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     2993..3012
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3231..3335)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3403..3421
                     /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 acccggggac
      421 aagctgctgc tttagaatta gttgatccta tggtctccaa gggtgaggag gacaacatgg
      481 cctccctccc cgccacccac gagttacaca tctttggttc catcaacggt gtcgacttcg
      541 acatggtcgg tcagggtacc ggtaacccca acgacggtta cgaggagctc aacctcaagt
      601 ccaccaaggg tgacctccag ttctccccct ggattctcgt ccctcatatc ggttacggtt
      661 tccaccagta cctcccctac cctgacggta tgtccccctt ccaggccgcc atggttgatg
      721 gttccggtta ccaggtccac cgcactatgc agtttgaaga tggtgcctcc ctcaccgtca
      781 actaccgcta cacctacgag ggttcccaca tcaaaggtga agcccaggtc aagggtaccg
      841 gtttccctgc tgacggtcct gtcatgacca actccctcac cgctgctgac tggtgccgtt
      901 ccaagaagac ttaccccaac gacaagacca tcatctccac cttcaagtgg tcctacacca
      961 ccggtaatgg taagcgctac cgctccaccg cccgcaccac ctacaccttt gccaagccta
     1021 tggccgccaa ctatctcaag aaccagccca tgtacgtctt ccgtaagacc gagctcaagc
     1081 actccaagac cgagctcaac ttcaaggagt ggcagaaggc cttcaccgac gtcatgggta
     1141 tggacgagct ctacaagtaa taatcctcta gagtcgacct gcaggcatgc aagcttggcg
     1201 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     1261 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     1321 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     1381 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     1441 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     1501 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     1561 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     1621 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     1681 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     1741 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     1801 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     1861 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     1921 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     1981 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     2041 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     2101 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     2161 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     2221 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     2281 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     2341 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     2401 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     2461 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     2521 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     2581 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     2641 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     2701 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     2761 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     2821 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     2881 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     2941 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     3001 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     3061 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     3121 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     3181 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     3241 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     3301 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     3361 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     3421 ccctttcgtc
//
