LOCUS       sequence-330738-        6768 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses hygR from the gdpA promoter with a trpC terminator. 
            Typically used in Aspergillus species as a selection marker for 
            resistance to hygromycin..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6768)
  AUTHORS   Zhao C, Fraczek MG, Dineen L, Lebedinec R, Macheleidt J, Heinekamp 
            T, Delneri D, Bowyer P, Brakhage AA, Bromley M
  TITLE     High-Throughput Gene Replacement in Aspergillus fumigatus.
  JOURNAL   Curr Protoc Microbiol. 2019 Sep;54(1):e88. doi: 10.1002/cpmc.88.
  PUBMED    31518064
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: "Curr Protoc
            Microbiol."; date: "2019-09"; volume: "54(1):e88. doi"; pages: " 
            10.1002/cpmc.88"
COMMENT     Sequence Label: pAN7.1
FEATURES             Location/Qualifiers
     source          1..6768
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     151..170
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     404..421
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    538..559
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(574..579,580..597,598..604)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    612..628
                     /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     617..639
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     636..652
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     636..652
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        661..2798
                     /gene="gpdA"
                     /label=gpdA promoter
                     /note="promoter from the Aspergillus nidulans 
                     glyceraldehyde-3-phosphate dehydrogenase gene (Punt et al.,
                     1990)"
     intron          2847..2962
                     /gene="gpdA"
                     /label=gpdA intron
                     /note="intron from the Aspergillus nidulans 
                     glyceraldehyde-3-phosphate dehydrogenase gene (Punt et al.,
                     1990)"
     CDS             2971..3990
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGYV
                     LRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPET
                     ELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQTV
                     MDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFGD
                     SQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGNF
                     DDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE"
     terminator      4006..4781
                     /gene="trpC"
                     /label=trpC terminator
                     /note="transcription terminator from the Aspergillus 
                     nidulans trpC gene (Punt et al., 1987)"
     primer_bind     complement(4804..4821)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4804..4820)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4813..4835)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5029..5048)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5148..5170
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5208..5226)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5294..5398
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5399..5467,5468..6259)
                     /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(5617..5636)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(6430..6768,1..250)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
       61 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
      121 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
      181 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
      241 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
      301 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
      361 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
      421 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
      481 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
      541 tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat
      601 gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg acatgattac
      661 gaattccctt gtatctctac acacaggctc aaatcaataa gaagaacggt tcgtcttttt
      721 cgtttatatc ttgcatcgtc ccaaagctat tggcgggata ttctgtttgc agttggctga
      781 cttgaagtaa tctctgcaga tctttcgaca ctgaaatacg tcgagcctgc tccgcttgga
      841 agcggcgagg agcctcgtcc tgtcacaact accaacatgg agtacgataa gggccagttc
      901 cgccagctca ttaagagcca gttcatgggc gttggcatga tggccgtcat gcatctgtac
      961 ttcaagtaca ccaaccctct tctgatccag tcgatcatcc cgctgaaggg cgctttcgaa
     1021 tcgaatctgg ttaagatcca cgtcttcggg aagccagcga ctggtgacct ccagcgtccc
     1081 tttaaggctg ccaacagctt tctcagccag ggccagccca agaccgacaa ggcctccctc
     1141 cagaacgccg agaagaactg gaggggtggt gtcaaggagg agtaagctcc ttattgaagt
     1201 cggaggacgg agcggtgtca agaggatatt cttcgctctg tattatagat aagatgatga
     1261 ggaattggag gtagcatagc ttcatttgga tttgctttcc aggctgagac tctagcttgg
     1321 agcatagagg gtccctttgg ctttcaatat tctcaagtat ctcgagtttg aacttattcc
     1381 cgtgaacctt ttattcacca atgagcattg gaatgaacat gaatctgagg actgcaatcg
     1441 ccatgaggtt ttcgaaatac atccggatgt cgaaggcttg gggcacctgc gttggttgaa
     1501 tttagaacgt ggcactattg atcatccgat agctctgcaa agggcgttgc acaatgcaag
     1561 tcaaacgttg ctagcagttc caggtggaat gttatgatga gcattgtatt aaatcaggag
     1621 atatagcatg atctctagtt agctcaccac aaaagtcaga cggcgtaacc aaaagtcaca
     1681 caacacaagc tgtaaggatt tcggcacggc tacggaagac ggagaagccc accttcagtg
     1741 gactcgagta ccatttaatt ctatttgtgt ttgatcgaga cctaatacag cccctacaac
     1801 gaccatcaaa gtcgtatagc taccagtgag gaagtggact caaatcgact tcagcaacat
     1861 ctcctggata aactttaagc ctaaactata cagaataaga tggtggagag cttataccga
     1921 gctcccaaat ctgtccagat catggttgac cggtgcctgg atcttcctat agaatcatcc
     1981 ttattcgttg acctagctga ttctggagtg acccagaggg tcatgacttg agcctaaaat
     2041 ccgccgcctc caccatttgt agaaaaatgt gacgaactcg tgagctctgt acagtgaccg
     2101 gtgactcttt ctggcatgcg gagagacgga cggacgcaga gagaagggct gagtaataag
     2161 cgccactgcg ccagacagct ctggcggctc tgaggtgcag tggatgatta ttaatccggg
     2221 accggccgcc cctccgcccc gaagtggaaa ggctggtgtg cccctcgttg accaagaatc
     2281 tattgcatca tcggagaata tggagcttca tcgaatcacc ggcagtaagc gaaggagaat
     2341 gtgaagccag gggtgtatag ccgtcggcga aatagcatgc cattaaccta ggtacagaag
     2401 tccaattgct tccgatctgg taaaagattc acgagatagt accttctccg aagtaggtag
     2461 agcgagtacc cggcgcgtaa gctccctaat tggcccatcc ggcatctgta gggcgtccaa
     2521 atatcgtgcc tctcctgctt tgcccggtgt atgaaaccgg aaaggccgct caggagctgg
     2581 ccagcggcgc agaccgggaa cacaagctgg cagtcgaccc atccggtgct ctgcactcga
     2641 cctgctgagg tccctcagtc cctggtaggc agctttgccc cgtctgtccg cccggtgtgt
     2701 cggcggggtt gacaaggtcg ttgcgtcagt ccaacatttg ttgccatatt ttcctgctct
     2761 ccccaccagc tgctcttttc ttttctcttt cttttcccat cttcagtata ttcatcttcc
     2821 catccaagaa cctttatttc ccctaagtaa gtactttgct acatccatac tccatccttc
     2881 ccatccctta ttcctttgaa cctttcagtt cgagctttcc cacttcatcg cagcttgact
     2941 aacagctacc ccgcttgagc agacatcacc atgcctgaac tcaccgcgac gtctgtcgag
     3001 aagtttctga tcgaaaagtt cgacagcgtc tccgacctga tgcagctctc ggagggcgaa
     3061 gaatctcgtg ctttcagctt cgatgtagga gggcgtggat atgtcctgcg ggtaaatagc
     3121 tgcgccgatg gtttctacaa agatcgttat gtttatcggc actttgcatc ggccgcgctc
     3181 ccgattccgg aagtgcttga cattggggaa ttcagcgaga gcctgaccta ttgcatctcc
     3241 cgccgtgcac agggtgtcac gttgcaagac ctgcctgaaa ccgaactgcc cgctgttctg
     3301 cagccggtcg cggaggccat ggatgcgatc gctgcggccg atcttagcca gacgagcggg
     3361 ttcggcccat tcggaccgca aggaatcggt caatacacta catggcgtga tttcatatgc
     3421 gcgattgctg atccccatgt gtatcactgg caaactgtga tggacgacac cgtcagtgcg
     3481 tccgtcgcgc aggctctcga tgagctgatg ctttgggccg aggactgccc cgaagtccgg
     3541 cacctcgtgc acgcggattt cggctccaac aatgtcctga cggacaatgg ccgcataaca
     3601 gcggtcattg actggagcga ggcgatgttc ggggattccc aatacgaggt cgccaacatc
     3661 ttcttctgga ggccgtggtt ggcttgtatg gagcagcaga cgcgctactt cgagcggagg
     3721 catccggagc ttgcaggatc gccgcggctc cgggcgtata tgctccgcat tggtcttgac
     3781 caactctatc agagcttggt tgacggcaat ttcgatgatg cagcttgggc gcagggtcga
     3841 tgcgacgcaa tcgtccgatc cggagccggg actgtcgggc gtacacaaat cgcccgcaga
     3901 agcgcggccg tctggaccga tggctgtgta gaagtactcg ccgatagtgg aaaccgacgc
     3961 cccagcactc gtccgagggc aaaggaatag agtagatgcc gaccgggatc cacttaacgt
     4021 tactgaaatc atcaaacagc ttgacgaatc tggatataag atcgttggtg tcgatgtcag
     4081 ctccggagtt gagacaaatg gtgttcagga tctcgataag atacgttcat ttgtccaagc
     4141 agcaaagagt gccttctagt gatttaatag ctccatgtca acaagaataa aacgcgtttc
     4201 gggtttacct cttccagata cagctcatct gcaatgcatt aatgcattgg acctcgcaac
     4261 cctagtacgc ccttcaggct ccggcgaagc agaagaatag cttagcagag tctattttca
     4321 ttttcgggag acgagatcaa gcagatcaac ggtcgtcaag agacctacga gactgaggaa
     4381 tccgctcttg gctccacgcg actatatatt tgtctctaat tgtactttga catgctcctc
     4441 ttctttactc tgatagcttg actatgaaaa ttccgtcacc agcccctggg ttcgcaaaga
     4501 taattgcact gtttcttcct tgaactctca agcctacagg acacacattc atcgtaggta
     4561 taaacctcga aaatcattcc tactaagatg ggtatacaat agtaaccatg catggttgcc
     4621 tagtgaatgc tccgtaacac ccaatacgcc ggccgaaact tttttacaac tctcctatga
     4681 gtcgtttacc cagaatgcac aggtacactt gtttagaggt aatccttctt tctagaagtc
     4741 ctcgtgtact gtgtaagcgc ccactccaca tctccactcg acctgcaggc atgcaagctt
     4801 ggcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa
     4861 tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga
     4921 tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg cgcctgatgc ggtattttct
     4981 ccttacgcat ctgtgcggta tttcacaccg catatggtgc actctcagta caatctgctc
     5041 tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg
     5101 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     5161 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg
     5221 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt
     5281 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     5341 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     5401 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     5461 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     5521 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     5581 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     5641 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     5701 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     5761 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     5821 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     5881 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     5941 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     6001 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     6061 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     6121 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     6181 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     6241 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     6301 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     6361 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     6421 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     6481 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     6541 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     6601 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     6661 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     6721 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacaca
//
