LOCUS       Exported                8276 bp DNA     circular SYN 15-MAY-2025
DEFINITION  Vector with a hygromycin B resistance marker for transforming 
            Aspergillus species.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8276)
  AUTHORS   Punt PJ, Oliver RP, Dingemanse MA, Pouwels PH, van den Hondel CA.
  TITLE     Transformation of Aspergillus based on the hygromycin B resistance 
            marker from Escherichia coli.
  JOURNAL   Gene 1987;56:117-24.
  PUBMED    2824287
REFERENCE   2  (bases 1 to 8276)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 8276)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8276)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 8276)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Gene"; 
            date: "1987"; volume: "56"; pages: "117-24"
COMMENT     SGRef: number: 2; type: "Journal Article"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     SGRef: number: 4; type: "Journal Article"
COMMENT     The trpC terminator region was corrected based on the updated 
            sequence record X02390.
FEATURES             Location/Qualifiers
     source          1..8276
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          3337..4858
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..2129
                     /label=gpdA promoter
                     /note="promoter from the Aspergillus nidulans 
                     glyceraldehyde-3-phosphate dehydrogenase gene (Punt et al.,
                     1990)"
     intron          2178..2293
                     /label=gpdA intron
                     /note="intron from the Aspergillus nidulans 
                     glyceraldehyde-3-phosphate dehydrogenase gene (Punt et al.,
                     1990)"
     CDS             2302..3318
                     /codon_start=1
                     /label=HygR
                     /note="aminoglycoside phosphotransferase from E. coli"
                     /translation="MPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGYV
                     LRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPET
                     ELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQTV
                     MDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFGD
                     SQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGNF
                     DDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE"
     CDS             4100..4816
                     /codon_start=1
                     /label=EGFP
                     /note="enhanced GFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4840..4856)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      4858..5628
                     /label=trpC terminator
                     /note="transcription terminator from the Aspergillus
                     nidulans trpC gene (Punt et al., 1987)"
     primer_bind     complement(5651..5667)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6141..6245
                     /label=AmpR promoter
     CDS             6246..7103
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      7277..7865
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    8153..8174
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        8189..8219
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8227..8243
                     /label=lac operator
                     /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     8251..8267
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gaattccctt gtatctctac acacaggctc aaatcaataa gaagaacggt tcgtcttttt
       61 cgtttatatc ttgcatcgtc ccaaagctat tggcgggata ttctgtttgc agttggctga
      121 cttgaagtaa tctctgcaga tctttcgaca ctgaaatacg tcgagcctgc tccgcttgga
      181 agcggcgagg agcctcgtcc tgtcacaact accaacatgg agtacgataa gggccagttc
      241 cgccagctca ttaagagcca gttcatgggc gttggcatga tggccgtcat gcatctgtac
      301 ttcaagtaca ccaacgctct tctgatccag tcgatcatcc gctgaaggcg ctttcgaatc
      361 tggttaagat ccacgtcttc gggaagccag cgactggtga cctccagcgt ccctttaagg
      421 ctgccaacag ctttctcagc cagggccagc ccaagaccga caaggcctcc ctccagaacg
      481 ccgagaagaa ctggaggggt ggtgtcaagg aggagtaagc tccttattga agtcggagga
      541 cggagcggtg tcaagaggat attcttcgac tctgtattat agataagatg atgaggaatt
      601 ggaggtagca tagcttcatt tggatttgct ttccaggctg agactctagc ttggagcata
      661 gagggtcctt tggctttcaa tattctcaag tatctcgagt ttgaacttat tccctgtgaa
      721 ccttttattc accaatgagc attggaatga acatgaatct gaggactgca atcgccatga
      781 ggttttcgaa atacatccgg atgtcgaagg cttggggcac ctgcgttggt tgaatttaga
      841 acgtggcact attgatcatc cgatagctct gcaaagggcg ttgcacaatg caagtcaaac
      901 gttgctagca gttccaggtg gaatgttatg atgagcattg tattaaatca ggagatatag
      961 catgatctct agttagctca ccacaaaagt cagacggcgt aaccaaaagt cacacaacac
     1021 aagctgtaag gatttcggca cggctacgga agacggagaa gccaccttca gtggactcga
     1081 gtaccattta attctatttg tgtttgatcg agacctaata cagcccctac aacgaccatc
     1141 aaagtcgtat agctaccagt gaggaagtgg actcaaatcg acttcagcaa catctcctgg
     1201 ataaacttta agcctaaact atacagaata agataggtgg agagcttata ccgagctccc
     1261 aaatctgtcc agatcatggt tgaccggtgc ctggatcttc ctatagaatc atccttattc
     1321 gttgacctag ctgattctgg agtgacccag agggtcatga cttgagccta aaatccgccg
     1381 cctccaccat ttgtagaaaa atgtgacgaa ctcgtgagct ctgtacagtg accggtgact
     1441 ctttctggca tgcggagaga cggacggacg cagagagaag ggctgagtaa taagccactg
     1501 gccagacagc tctggcggct ctgaggtgca gtggatgatt attaatccgg gaccggccgc
     1561 ccctccgccc cgaagtggaa aggctggtgt gcccctcgtt gaccaagaat ctattgcatc
     1621 atcggagaat atggagcttc atcgaatcac cggcagtaag cgaaggagaa tgtgaagcca
     1681 ggggtgtata gccgtcggcg aaatagcatg ccattaacct aggtacagaa gtccaattgc
     1741 ttccgatctg gtaaaagatt cacgagatag taccttctcc gaagtaggta gagcgagtac
     1801 ccggcgcgta agctccctaa ttggcccatc cggcatctgt agggcgtcca aatatcgtgc
     1861 ctctcctgct ttgcccggtg tatgaaaccg gaaaggccgc tcaggagctg gccagcggcg
     1921 cagaccggga acacaagctg gcagtcgacc catccggtgc tctgcactcg acctgctgag
     1981 gtccctcagt ccctggtagg cagctttgcc ccgtctgtcc gcccggtgtg tcggcggggt
     2041 tgacaaggtc gttgcgtcag tccaacattt gttgccatat tttcctgctc tccccaccag
     2101 ctgctctttt cttttctctt tcttttccca tcttcagtat attcatcttc ccatccaaga
     2161 acctttattt cccctaagta agtactttgc tacatccata ctccatcctt cccatccctt
     2221 attcctttga acctttcagt tcgagctttc ccacttcatc gcagcttgac taacagctac
     2281 cccgcttgag cagacatcac catgcctgaa ctcaccgcga cgtctgtcga gaagtttctg
     2341 atcgaaaagt tcgacagcgt ctccgacctg atgcagctct cggagggcga agaatctcgt
     2401 gctttcagct tcgatgtagg agggcgtgga tatgtcctgc gggtaaatag ctgcgccgat
     2461 ggtttctaca aagatcgtta tgtttatcgg cactttgcat cggccgcgct cccgattccg
     2521 gaagtgcttg acattgggga attcagcgag agcctgacct attgcatctc ccgccgtgca
     2581 cagggtgtca cgttgcaaga cctgcctgaa accgaactgc ccgctgttct gcagccggtc
     2641 gcggaggcca tggatgcgat cgctgcggcc gatcttagcc agacgagcgg gttcggccca
     2701 ttcggaccgc aaggaatcgg tcaatacact acatggcgtg atttcatatg cgcgattgct
     2761 gatccccatg tgtatcactg gcaaactgtg atggacgaca ccgtcagtgc gtccgtcgcg
     2821 caggctctcg atgagctgat gctttgggcc gaggactgcc ccgaagtccg gcacctcgtg
     2881 cacgcggatt tcggctccaa caatgtcctg acggacaatg gccgcataac agcggtcatt
     2941 gactggagcg aggcgatgtt cggggattcc caatacgagg tcgccaacat cttcttctgg
     3001 aggccgtggt tggcttgtat ggagcagcag acgcgctact tcgagcggag gcatccggag
     3061 cttgcaggat cgccgcggct ccgggcgtat atgctccgca ttggtcttga ccaactctat
     3121 cagagcttgg ttgacggcaa tttcgatgat gcagcttggg cgcagggtcg atgcgacgca
     3181 atcgtccgat ccggagccgg gactgtcggg cgtacacaaa tcgcccgcag aagcgcggcc
     3241 gtctggaccg atggctgtgt agaagtactc gccgatagtg gaaaccgacg ccccagcact
     3301 cgtccgaggg caaaggaata gagtagatgc cgaccgcggc agaagtatgc aaagcatgcg
     3361 gagagacgga cggacgcaga gagaagggct gagtaataag cgccactgcg ccagacagct
     3421 ctggcggctc tgaggtgcag tggatgatta ttaatccggg accggccgcc cctccgcccc
     3481 gaagtggaaa ggctggtgtg cccctcgttg accaagaatc tattgcatca tcggagaata
     3541 tggagcttca tcgaatcacc ggcagtaagc gaaggagaat gtgaagccag gggtgtatag
     3601 ccgtcggcga aatagcatgc cattaaccta ggtacagaag tccaattgct tccgatctgg
     3661 taaaagattc acgagatagt accttctccg aagtaggtag agcgagtacc cggcgcgtaa
     3721 gctccctaat tggcccatcc ggcatctgta gggcgtccaa atatcgtgcc tctcctgctt
     3781 tgcccggtgt atgaaaccgg aaaggccgct caggagctgg ccagcggcgc agaccgggaa
     3841 cacaagctgg cagtcgaccc atccggtgct ctgcactcga cctgctgagg tccctcagtc
     3901 cctggtaggc agctttgccc cgtctgtccg cccggtgtgt cggcggggtt gacaaggtcg
     3961 ttgcgtcagt ccaacatttg ttgccatatt ttcctgctct ccccaccagc tgctcttttc
     4021 ttttctcttt cttttcccat cttcagtata ttcatcttcc catccaagaa cctttatcga
     4081 cctcgactct agaggatcca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
     4141 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
     4201 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct
     4261 gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg
     4321 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
     4381 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
     4441 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
     4501 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcat
     4561 ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga
     4621 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
     4681 gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga
     4741 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
     4801 ggacgagctg tacaagtaag aattcgatat caagcttatc gataccgtcg acctcgagac
     4861 ttaacgttac tgaaatcatc aaacagcttg acgaatctgg atataagatc gttggtgtcg
     4921 atgtcagctc cggagttgag acaaatggtg ttcaggatct cgataagata cgttcatttg
     4981 tccaagcagc aaagagtgcc ttctagtgat ttaatagctc catgtcaaca agaataaaac
     5041 gcgtttcggg tttacctctt ccagatacag ctcatctgca atgcattaat gcattggacc
     5101 tcgcaaccct agtacgccct tcaggctccg gcgaagcaga agaatagctt agcagagtct
     5161 attttcattt tcgggagacg agatcaagca gatcaacggt cgtcaagaga cctacgagac
     5221 tgaggaatcc gctcttggct ccacgcgact atatatttgt ctctaattgt actttgacat
     5281 gctcctcttc tttactctga tagcttgact atgaaaattc cgtcaccagc ccctgggttc
     5341 gcaaagataa ttgcactgtt tcttccttga actctcaagc ctacaggaca cacattcatc
     5401 gtaggtataa acctcgaaaa tcattcctac taagatgggt atacaatagt aaccatgcat
     5461 ggttgcctag tgaatgctcc gtaacaccca atacgccggc cgaaactttt ttacaactct
     5521 cctatgagtc gtttacccag aatgcacagg tacacttgtt tagaggtaat ccttctttct
     5581 agaagtcctc gtgtactgtg taagcgccca ctccacatct ccactcgacc tgcaggcatg
     5641 caagcttggc actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc
     5701 aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc
     5761 gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc ctgatgcggt
     5821 attttctcct tacgcatctg tgcggtattt cacaccgcat atggtgcact ctcagtacaa
     5881 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc
     5941 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     6001 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg
     6061 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     6121 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     6181 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     6241 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     6301 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     6361 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     6421 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     6481 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     6541 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     6601 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     6661 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     6721 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     6781 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     6841 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     6901 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     6961 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     7021 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     7081 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     7141 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     7201 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     7261 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     7321 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     7381 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     7441 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     7501 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     7561 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     7621 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     7681 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     7741 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     7801 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     7861 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     7921 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     7981 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     8041 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     8101 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
     8161 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
     8221 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat gattac
//
