LOCUS       sequence-370801-        6768 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pUC19 plasmid (MCS intact) with Hygromycin selection marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6768)
  AUTHORS   Storer ISR, Sastre-Velasquez LE, Easter T, Mertens B, Dallemulle A, 
            Bottery M, Tank R, Offterdinger M, Bromley MJ, van Rhijn N, Gsaller 
            F
  TITLE     Shining a light on the impact of antifungals on Aspergillus 
            fumigatus subcellular dynamics through fluorescence imaging.
  JOURNAL   Antimicrob Agents Chemother. 2024 Nov 6;68(11):e0080324. doi: 
            10.1128/aac.00803-24. Epub 2024 Oct 15.
  PUBMED    39404344
REFERENCE   2  (bases 1 to 6768)
  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: "Antimicrob 
            Agents Chemother."; date: "2024-11-6"; volume: "68(11):e0080324. 
            doi"; pages: " 10.1128/aac.00803-24. Epub 2024 Oct 15"
COMMENT     Sequence Label: pUC19-Hph
FEATURES             Location/Qualifiers
     source          1..6768
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          310..425
                     /gene="gpdA"
                     /label=gpdA intron
                     /note="intron from the Aspergillus nidulans 
                     glyceraldehyde-3-phosphate dehydrogenase gene (Punt et al.,
                     1990)"
     CDS             434..1453
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGYV
                     LRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPET
                     ELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQTV
                     MDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFGD
                     SQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGNF
                     DDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE"
     terminator      1469..2244
                     /gene="trpC"
                     /label=trpC terminator
                     /note="transcription terminator from the Aspergillus 
                     nidulans trpC gene (Punt et al., 1987)"
     primer_bind     complement(2267..2284)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2267..2283)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2276..2298)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2492..2511)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2611..2633
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2671..2689)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2757..2861
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2862..2930,2931..3722)
                     /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(3080..3099)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3893..4481
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4382..4401
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4635..4652
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4769..4790
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4805..4810,4811..4828,4829..4835)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4843..4859
                     /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     4848..4870
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4867..4883
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4867..4883
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        join(4892..6768,1..261)
                     /gene="gpdA"
                     /label=gpdA promoter
                     /note="promoter from the Aspergillus nidulans 
                     glyceraldehyde-3-phosphate dehydrogenase gene (Punt et al.,
                     1990)"
ORIGIN
        1 ctttgcccgg tgtatgaaac cggaaaggcc gctcaggagc tggccagcgg cgcagaccgg
       61 gaacacaagc tggcagtcga cccatccggt gctctgcact cgacctgctg aggtccctca
      121 gtccctggta ggcagctttg ccccgtctgt ccgcccggtg tgtcggcggg gttgacaagg
      181 tcgttgcgtc agtccaacat ttgttgccat attttcctgc tctccccacc agctgctctt
      241 ttcttttctc tttcttttcc catcttcagt atattcatct tcccatccaa gaacctttat
      301 ttcccctaag taagtacttt gctacatcca tactccatcc ttcccatccc ttattccttt
      361 gaacctttca gttcgagctt tcccacttca tcgcagcttg actaacagct accccgcttg
      421 agcagacatc accatgcctg aactcaccgc gacgtctgtc gagaagtttc tgatcgaaaa
      481 gttcgacagc gtctccgacc tgatgcagct ctcggagggc gaagaatctc gtgctttcag
      541 cttcgatgta ggagggcgtg gatatgtcct gcgggtaaat agctgcgccg atggtttcta
      601 caaagatcgt tatgtttatc ggcactttgc atcggccgcg ctcccgattc cggaagtgct
      661 tgacattggg gaattcagcg agagcctgac ctattgcatc tcccgccgtg cacagggtgt
      721 cacgttgcaa gacctgcctg aaaccgaact gcccgctgtt ctgcagccgg tcgcggaggc
      781 catggatgcg atcgctgcgg ccgatcttag ccagacgagc gggttcggcc cattcggacc
      841 gcaaggaatc ggtcaataca ctacatggcg tgatttcata tgcgcgattg ctgatcccca
      901 tgtgtatcac tggcaaactg tgatggacga caccgtcagt gcgtccgtcg cgcaggctct
      961 cgatgagctg atgctttggg ccgaggactg ccccgaagtc cggcacctcg tgcacgcgga
     1021 tttcggctcc aacaatgtcc tgacggacaa tggccgcata acagcggtca ttgactggag
     1081 cgaggcgatg ttcggggatt cccaatacga ggtcgccaac atcttcttct ggaggccgtg
     1141 gttggcttgt atggagcagc agacgcgcta cttcgagcgg aggcatccgg agcttgcagg
     1201 atcgccgcgg ctccgggcgt atatgctccg cattggtctt gaccaactct atcagagctt
     1261 ggttgacggc aatttcgatg atgcagcttg ggcgcagggt cgatgcgacg caatcgtccg
     1321 atccggagcc gggactgtcg ggcgtacaca aatcgcccgc agaagcgcgg ccgtctggac
     1381 cgatggctgt gtagaagtac tcgccgatag tggaaaccga cgccccagca ctcgtccgag
     1441 ggcaaaggaa tagagtagat gccgaccggg atccacttaa cgttactgaa atcatcaaac
     1501 agcttgacga atctggatat aagatcgttg gtgtcgatgt cagctccgga gttgagacaa
     1561 atggtgttca ggatctcgat aagatacgtt catttgtcca agcagcaaag agtgccttct
     1621 agtgatttaa tagctccatg tcaacaagaa taaaacgcgt ttcgggttta cctcttccag
     1681 atacagctca tctgcaatgc attaatgcat tggacctcgc aaccctagta cgcccttcag
     1741 gctccggcga agcagaagaa tagcttagca gagtctattt tcattttcgg gagacgagat
     1801 caagcagatc aacggtcgtc aagagaccta cgagactgag gaatccgctc ttggctccac
     1861 gcgactatat atttgtctct aattgtactt tgacatgctc ctcttcttta ctctgatagc
     1921 ttgactatga aaattccgtc accagcccct gggttcgcaa agataattgc actgtttctt
     1981 ccttgaactc tcaagcctac aggacacaca ttcatcgtag gtataaacct cgaaaatcat
     2041 tcctactaag atgggtatac aatagtaacc atgcatggtt gcctagtgaa tgctccgtaa
     2101 cacccaatac gccggccgaa acttttttac aactctccta tgagtcgttt acccagaatg
     2161 cacaggtaca cttgtttaga ggtaatcctt ctttctagaa gtcctcgtgt actgtgtaag
     2221 cgcccactcc acatctccac tcgacctgca ggcatgcaag cttggcactg gccgtcgttt
     2281 tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc
     2341 cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt
     2401 tgcgcagcct gaatggcgaa tggcgcctga tgcggtattt tctccttacg catctgtgcg
     2461 gtatttcaca ccgcatatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     2521 gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     2581 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     2641 cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta
     2701 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     2761 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     2821 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     2881 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     2941 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     3001 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     3061 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     3121 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     3181 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     3241 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     3301 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     3361 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     3421 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     3481 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     3541 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     3601 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     3661 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     3721 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     3781 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     3841 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     3901 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     3961 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     4021 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     4081 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     4141 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     4201 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     4261 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     4321 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     4381 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     4441 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     4501 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     4561 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     4621 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac
     4681 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
     4741 ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc
     4801 aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat
     4861 ttcacacagg aaacagctat gacatgatta cgaattccct tgtatctcta cacacaggct
     4921 caaatcaata agaagaacgg ttcgtctttt tcgtttatat cttgcatcgt cccaaagcta
     4981 ttggcgggat attctgtttg cagttggctg acttgaagta atctctgcag atctttcgac
     5041 actgaaatac gtcgagcctg ctccgcttgg aagcggcgag gagcctcgtc ctgtcacaac
     5101 taccaacatg gagtacgata agggccagtt ccgccagctc attaagagcc agttcatggg
     5161 cgttggcatg atggccgtca tgcatctgta cttcaagtac accaaccctc ttctgatcca
     5221 gtcgatcatc ccgctgaagg gcgctttcga atcgaatctg gttaagatcc acgtcttcgg
     5281 gaagccagcg actggtgacc tccagcgtcc ctttaaggct gccaacagct ttctcagcca
     5341 gggccagccc aagaccgaca aggcctccct ccagaacgcc gagaagaact ggaggggtgg
     5401 tgtcaaggag gagtaagctc cttattgaag tcggaggacg gagcggtgtc aagaggatat
     5461 tcttcgctct gtattataga taagatgatg aggaattgga ggtagcatag cttcatttgg
     5521 atttgctttc caggctgaga ctctagcttg gagcatagag ggtccctttg gctttcaata
     5581 ttctcaagta tctcgagttt gaacttattc ccgtgaacct tttattcacc aatgagcatt
     5641 ggaatgaaca tgaatctgag gactgcaatc gccatgaggt tttcgaaata catccggatg
     5701 tcgaaggctt ggggcacctg cgttggttga atttagaacg tggcactatt gatcatccga
     5761 tagctctgca aagggcgttg cacaatgcaa gtcaaacgtt gctagcagtt ccaggtggaa
     5821 tgttatgatg agcattgtat taaatcagga gatatagcat gatctctagt tagctcacca
     5881 caaaagtcag acggcgtaac caaaagtcac acaacacaag ctgtaaggat ttcggcacgg
     5941 ctacggaaga cggagaagcc caccttcagt ggactcgagt accatttaat tctatttgtg
     6001 tttgatcgag acctaataca gcccctacaa cgaccatcaa agtcgtatag ctaccagtga
     6061 ggaagtggac tcaaatcgac ttcagcaaca tctcctggat aaactttaag cctaaactat
     6121 acagaataag atggtggaga gcttataccg agctcccaaa tctgtccaga tcatggttga
     6181 ccggtgcctg gatcttccta tagaatcatc cttattcgtt gacctagctg attctggagt
     6241 gacccagagg gtcatgactt gagcctaaaa tccgccgcct ccaccatttg tagaaaaatg
     6301 tgacgaactc gtgagctctg tacagtgacc ggtgactctt tctggcatgc ggagagacgg
     6361 acggacgcag agagaagggc tgagtaataa gcgccactgc gccagacagc tctggcggct
     6421 ctgaggtgca gtggatgatt attaatccgg gaccggccgc ccctccgccc cgaagtggaa
     6481 aggctggtgt gcccctcgtt gaccaagaat ctattgcatc atcggagaat atggagcttc
     6541 atcgaatcac cggcagtaag cgaaggagaa tgtgaagcca ggggtgtata gccgtcggcg
     6601 aaatagcatg ccattaacct aggtacagaa gtccaattgc ttccgatctg gtaaaagatt
     6661 cacgagatag taccttctcc gaagtaggta gagcgagtac ccggcgcgta agctccctaa
     6721 ttggcccatc cggcatctgt agggcgtcca aatatcgtgc ctctcctg
//
