LOCUS       sequence-429382-        3421 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Aspergillus nidulans codon-adjusted mNeonGreen fluorescent protein, 
            includes linker for N-terminal tagging..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3421)
  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 3421)
  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: pDM053
FEATURES             Location/Qualifiers
     source          1..3421
                     /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      678..687
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(1200..1216)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1200..1216)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1213..1235)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1224..1240
                     /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(1248..1254,1255..1272,1273..1278))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1293..1314
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1431..1448)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1602..2190)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1682..1701)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2361..3152,3153..3221))
                     /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     2984..3003
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3222..3326)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3394..3412
                     /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 gaggacaaca tggcctccct ccccgccacc cacgagttac
      481 acatctttgg ttccatcaac ggtgtcgact tcgacatggt cggtcagggt accggtaacc
      541 ccaacgacgg ttacgaggag ctcaacctca agtccaccaa gggtgacctc cagttctccc
      601 cctggattct cgtccctcat atcggttacg gtttccacca gtacctcccc taccctgacg
      661 gtatgtcccc cttccaggcc gccatggttg atggttccgg ttaccaggtc caccgcacta
      721 tgcagtttga agatggtgcc tccctcaccg tcaactaccg ctacacctac gagggttccc
      781 acatcaaagg tgaagcccag gtcaagggta ccggtttccc tgctgacggt cctgtcatga
      841 ccaactccct caccgctgct gactggtgcc gttccaagaa gacttacccc aacgacaaga
      901 ccatcatctc caccttcaag tggtcctaca ccaccggtaa tggtaagcgc taccgctcca
      961 ccgcccgcac cacctacacc tttgccaagc ctatggccgc caactatctc aagaaccagc
     1021 ccatgtacgt cttccgtaag accgagctca agcactccaa gaccgagctc aacttcaagg
     1081 agtggcagaa ggccttcacc gacgtcatgg gtatggacga gctctacaag aacatcctcg
     1141 gtcccatccg cggatcctct agagtcgacc tgcaggcatg caagcttggc gtaatcatgg
     1201 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     1261 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     1321 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     1381 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     1441 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     1501 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     1561 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     1621 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     1681 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     1741 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     1801 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     1861 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     1921 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     1981 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     2041 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     2101 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     2161 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     2221 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     2281 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     2341 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     2401 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     2461 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     2521 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     2581 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     2641 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     2701 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     2761 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     2821 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     2881 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     2941 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     3001 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     3061 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     3121 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     3181 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     3241 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     3301 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
     3361 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
     3421 c
//
