LOCUS       sequence-458622-       10316 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Heterologous production of pleuromutilin in Aspergillus oryzae.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10316)
  TITLE     Aspergillus oryzae plasmids
REFERENCE   2  (bases 1 to 10316)
  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"
COMMENT     Sequence Label: ppG-ple12
FEATURES             Location/Qualifiers
     source          1..10316
                     /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"
     protein_bind    2049..2065
                     /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     2073..2089
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4487..4503
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8095..8111)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    8119..8135
                     /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(8143..8149,8150..8167,8168..8173))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8188..8209
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8326..8343)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8497..9085)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8577..8596)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9256..10047,10048..10116))
                     /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     9879..9898
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10117..10221)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     10289..10307
                     /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 acccaatcct
      421 gcctacctaa catgggattt agtcattcct tttggcatca aagcaaggcg ccacaatcgt
      481 tgaggtaaga atgatctgcc aatatgtgag cgagtgcggc tgacaactat gatagtttgg
      541 taatacaaca tctcacgaat cgtaccttgc atgtgcacta aagtttagtc agatgtacag
      601 ttaacaagag atctcgtgcc cgtcatatac catgactttt ctctgagcga gtcagggact
      661 gatatcccta ttcatgactt aaatcttgat caggtcagta tctttgagat tgatttgaac
      721 tgattctcac ccatcaaagt tcatgcacgc aagcaatatc cagtcgccac gaggggatcc
      781 cgtatctgtg cttgggaaag cgaatgcaca gcctatatcc actcaggtta cttcgactaa
      841 gccacgatct cgatcattaa ctaaagacca cgagagggga actagggaaa tccgtgatag
      901 aatgaagtat acagttgatt ttgtcagcaa gggattcaaa cctaatacca gaggtgactt
      961 tatccaagat tccttcacga cactggaaga actgcttgaa gagctgcctg agtcaatcag
     1021 cttcaatatc gagataagta ggttatatgc tgccactggt ggttctccac ttgcgggaga
     1081 caaagctaac aacgtcccaa tgaagagtac cccaggcttc atgaagctat agaagcaggt
     1141 gtagcaccag tggctattga aatcaacacc ttcatcgaca aagcgcttga gagactcttt
     1201 tcttacggca acaaaaaacg gaccattatc ctatcctcat ttactcccga gatctgcatt
     1261 ttattggcca tcaaacaaca gacgtaccct gtgatgttca tcactaatgc cggcaagcct
     1321 ccagttacgg atcgagagat gagggctgcc agcatacagt ccgctgttcg atttgccaag
     1381 aggtggaatt tatctggcct tgtctttgca tctgaggcgc tggtaatgtg ccccaggctt
     1441 gtcagatatg ttcaacgatc aggattgatc tgtggatcct atggatctca gaacaatata
     1501 ccagaaaatg cgaaggtaag tgcttctata ttgatcctta gtgctttcaa actgtgatgt
     1561 agaagttgct cggtagctga ttaaatattc tagacccaag ccgctgctgg aattgacatt
     1621 attatggccg atagggttgg gcttattgct atgtccctga aaggatatca aaagcaggca
     1681 aaaagccagg cataatcccc gcgtggacgg taccctaagg ataggcccta atcttatcta
     1741 catgtgactg catcgatgtg tttggtcaaa atgaggcatg tggctcaccc cacaggcgga
     1801 gaaacgtgtg gctagtgcat gacagtcccc tccatagatt caatttaatt tttcgcggca
     1861 attgtcgtgc agtttgtatc tacatttcat tccatatatc aagagttagt agttggacat
     1921 cctgattatt ttgtctaatt actgaaaact cgaagtacta acctactaat aagccagttt
     1981 caaccactaa gtgctcattt atacaatatt tgcagaaccc cgcgctaccc ctccatcgcc
     2041 aaccccaatt gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacga
     2101 tatcatggtg ttttgatcat tttaaatttt tatatggcgg gtggtgggca actcgcttgc
     2161 gcgggcaact cgcttaccga ttacgttagg gctgatattt acgtaaaaat cgtcaaggga
     2221 tgcaagacca aagtagtaaa accccggagt caacagcatc caagcccaag tccttcacgg
     2281 agaaacccca gcgtccacat cacgagcgaa ggaccacctc taggcatcgg acgcaccatc
     2341 caattagaag cagcaaagcg aaacagccca agaaaaaggt cggcccgtcg gccttttctg
     2401 caacgctgat cacgggcagc gatccaacca acaccctcca gagtgactag gggcggaaat
     2461 ttaaagggat taatttccac tcaaccacaa atcacagtcg tccccggtat tgtcctgcag
     2521 aatgcaattt aaactcttct gcgaatcgct tggattcccc gcccctggcc gtagagctta
     2581 aagtatgtcc cttgtcgatg cgatgtatca caacatataa atactagcaa gggatgccat
     2641 gcttggagga tagcaaccga caacatcaca tcaagctctc ccttctctga acaataaacc
     2701 ccacagcaag ctccgaattc gagctcgatg gcaccgtcta atacagaacg tgctctaccg
     2761 gtccttgcaa tatgggccgc tataggcttg gcctactgga tagactctca aaagaagaaa
     2821 aagcaccact tgccgcctgg gccgaagaag cttccaatca ttggcaatgt tatggaccta
     2881 ccagcgaaaa tcgaatggga aacctacgct cgctggggta aagaatacaa ttcagacatc
     2941 attcaagtta gcgctgtcgg aacctccatc gtgattctga attcagccaa tgctgccaat
     3001 gatttgttac tgaagaggtc ggcgatttac tcgagcaggc cacacagcac aatgcaccac
     3061 gaactgtcag gatggggttt tacgtgggcg ttgatgccat acggcgagtc gtggcgggct
     3121 ggtcgaagaa gcttcaccaa gcacttcaac tcttcaaatc ctggaataaa ccaacctcgc
     3181 gagttgcgat acgtgaaacg cttcctcaag caactttacg agaagcccaa cgatgttctc
     3241 gaccatgtac ggaatctggt cggctctacg acgctttcta tgacctatgg gctcgagacg
     3301 gaaccctaca acgatcccta cgttgaactg gttgagaaag ccgtccttgc agcgtctgag
     3361 atcatgacct ctggagcctt tctcgtcgat atcatccccg cgatgaaaca cattcctcca
     3421 tgggtcccag ggactatctt ccaccagaag gctgctttaa tgcgaggaca tgcatactat
     3481 gttcgtgagc agcctttcaa agttgcccag gagatgataa aaaccggcga ctatgagccc
     3541 tcctttgtct ccgatgccct tcgagatctt gagaactcgg aaaccccaga ggaagatctg
     3601 gaacacctca aggatgtcgc tggtcaagtc tacatcgctg gcgctgatac gactgcatcc
     3661 gctctgggta ccttcttcct tgccatggtc tgcttcccgg aagtgcagaa gaaagcgcaa
     3721 caagagcttg atagtgttct caacggaagg ctgcccgagc acgctgactt cccgtctttc
     3781 ccatatctca acgcagtgat caaggaggtt taccgctgga gacctgtgac tcctatgggc
     3841 gtacctcatc aaactatcgc agatgacgtt tatagggact accatatccc taagggatcc
     3901 atcgtgttcg ctaaccaatg ggccatgtcc aacgacgaga atgattatcc ccagcccggc
     3961 gaattccggc ctgagcgata cttgactgaa gacggcaagc ctaacaaagc tgtcagagac
     4021 cccttcgata ttgcgttcgg cttcggcaga agaatttgtg ctggtcgtta tcttgctcat
     4081 tccacaatca ccttggccgc tgcttctgtt ttatctctct ttgatctctt aaaagcggtt
     4141 gacgaaaatg gcaaggagat cgagcctacc agagaatatc atcaggctat gatatcacgt
     4201 ccactagagt tcccgtgtcg tatcaagcca agaaataaag aagctgagga ggtcatccgt
     4261 gcctgcccgt tgacgttcac aaagcctacc agcgcttagc cggggatctg tagtagctcg
     4321 tgaagggtgg agagtatatg atggtactgc tattcaatct ggcattggac agtgagtttg
     4381 agtttgatgt acagttggag tcgttactgc tgtcatcccc ttatactctt cgattgtttt
     4441 tcgaacccta acgccaagca cgctagtcta ttataggaaa ggatcccagg aaacagctat
     4501 gaccatgatt acgatatcat ggtgttttga tcattttaaa tttttatatg gcgggtggtg
     4561 ggcaactcgc ttgcgcgggc aactcgctta ccgattacgt tagggctgat atttacgtaa
     4621 aaatcgtcaa gggatgcaag accaaagtag taaaaccccg gagtcaacag catccaagcc
     4681 caagtccttc acggagaaac cccagcgtcc acatcacgag cgaaggacca cctctaggca
     4741 tcggacgcac catccaatta gaagcagcaa agcgaaacag cccaagaaaa aggtcggccc
     4801 gtcggccttt tctgcaacgc tgatcacggg cagcgatcca accaacaccc tccagagtga
     4861 ctaggggcgg aaatttaaag ggattaattt ccactcaacc acaaatcaca gtcgtccccg
     4921 gtattgtcct gcagaatgca atttaaactc ttctgcgaat cgcttggatt ccccgcccct
     4981 ggccgtagag cttaaagtat gtcccttgtc gatgcgatgt atcacaacat ataaatacta
     5041 gcaagggatg ccatgcttgg aggatagcaa ccgacaacat cacatcaagc tctcccttct
     5101 ctgaacaata aaccccacag caagctccga attcgaatcg atttgagcta gcggccgcac
     5161 tagatgaagc ccttctcacc agaactactt gtcctatcct tcatcttatt ggttctttct
     5221 tgtgccatcc ggcctgctaa aggacgatgg atcctttggg tcatcatcgt tgcactcaac
     5281 acgtacctca ccatgaccac gaccggcgat tcgaccttgg actatgacat cgccaataac
     5341 ctcttcgtta tcactctcac cgctacggat tacattctct tgaccgatgt ccagagagag
     5401 ttgcaattcc gcaaccaaaa aggtgtcgag caagcctcgt tgcttgaacg catcaagtgg
     5461 gcgacatggc tcgtgcaaag tcggcgtgga gtgggctgga attgggagcc gaagattttc
     5521 gtccataggt tctccccgaa gacttcacgt ctttcattcc ttctccagca acttgtcaca
     5581 ggtgctcggc attacctcat ttgcgattta gtttcgctgt acagccgaag tccggtcgca
     5641 ttcgctgagc cccttgcttc tcgccctttg atctggcggt gtgcagatat tgccgcatgg
     5701 ctcctgttta ccaccaacca agtatcaatt cttcttacgg cattgagtct catgcaggtt
     5761 ctctctgggt actccgagcc gcaggactgg gttcccgtgt ttggccgctg gagagatgca
     5821 tacaccgttc ggcggttctg gggtcgatcg tggcatcagt tagttcgcag gtgtctatca
     5881 tcccccggaa aatacctttc tacgaaggtt ctaggcctga aacccggttc taaccccgcg
     5941 ctttacgtac aactgtacgc tgccttcttc ctttcgggag ttttgcacgc aattggggac
     6001 ttcaaggttc acgaggattg gtacaaggcc ggcaccatgg agttcttctg cgttcaagcg
     6061 gtgattatac agatggagga cggcgttctg tgggttggga ggaagcttgg tattaaggag
     6121 acctggtatt ggagggccct cggacatctt tggaccgtgg cgtggttcgt ctatagctgc
     6181 ccgaattggc taggggcaac tatctcagga aggggcaagg cctctatggc gttggaaagt
     6241 agtctcgtcc ttggtcttta ccggggagaa tggcaccctc ctcgcgtagc acaatagcta
     6301 gtgacccggg tagtcgtacc cgatgatgaa acataatgga caagttggtg aaaggttggg
     6361 ccagaggttg gacggtcgtg agggttatat tggggacggt gaatatatat aggaacatcg
     6421 ttccttatag tgcggatggt gtggtaagta tagactttgg atgaaattcg tttttggata
     6481 attgtgtcag tatatgattt ggtgtggtct tttcgtttga ttttagcata ggtgttatgg
     6541 gacgttaaga ctcttccacc aagaataaat gtattcttgt atactacaaa agtaccaaaa
     6601 tactccaatg aggagtctcg gttaggataa ctaaccagaa cggcgggcaa tctccaacat
     6661 agacgagaat ttgtaccgga gtgtctgaag gtgcagctgg agattggtct attttcttag
     6721 agtggggtat cactaatgta cagtcggtca ctatcgtaca aacaatcaca attatataca
     6781 agatttccca ccacccccta ctctaacacg gcacaattat ccatcgagtc agagcctagc
     6841 caccatttgg tgctctcgta gagaccaaag tataatcctg atccgacagc ggccataaac
     6901 gtgttgatag cacaccctcg gaatagtcct ctcgggccat ctgttcgtac aatctcccgt
     6961 acggtattga tcatcctttt cttctgaggt gcagttgtat ctgcagcatc gagcatgatt
     7021 cgtgtccgga ccatatccat gggtgctgtc aagacactag ctataccgcc cgagaccgca
     7081 gcacttattg cggctgtcgc tgcagcctct ccgattgtcg aatgggcctc tttctttcca
     7141 tactctcttg gtctttctag caccttctct cgatctccga atctatattc aaaaattcga
     7201 taccgaaaag actcgtacag aggcatctga atcgccgaca ctggcaagct atgcgccaca
     7261 agagccgggt atccgctcca aagctgtcta gggttgataa acttcttgaa agctagccgt
     7321 gtcgctttct gggctacacc acctaccctt cccccagcta caggtgctga tgcgtctgga
     7381 tggtgtgatt ggatcatctg cgcgttgtgt tttaatgcat cagccggagc aaagactccg
     7441 caagcagcaa gatccgcaac ggaggctgcg caaaaatcgg aaaagagccg agctgagcta
     7501 gactcatgcg ttccaagttt ttgatgtatg acttggagtc ctgactgtgc atactcgtat
     7561 gtgatgaaga atgcgcccgc tttgacccgt tattttttgc tcaatgaata tgagttgggt
     7621 aaggataatg ttgggtatca ccaacctgtg ggaaatgagg cagcggttac gctcgcgata
     7681 ccttggtaaa ggccgcggaa tatccctggg tgtctccata tgctggttcc tgtgttggtt
     7741 ctcaggaact gcgaatattc gcgagactga atgcgggttt tgatcgtatc taagggatat
     7801 gtgaatagat ctagtgagag ggatgcggtt gtgcttgcct agttccctca ggttaatgta
     7861 tgctgtttgt ccaagggggt tagaaaatat accagcaata tactaagtgg ctcggcggcc
     7921 atattatttg tcgatagagg cgggaggacg aggatgttca atcgtggatt ttgtaaccgt
     7981 cctggcacac atcaccttac aaggctttga gttttcgttc tataatttga gacaggtcac
     8041 agtcggggat cctctagagt cgacctgcag gcatgcaagc ttggcgtaat catggtcata
     8101 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     8161 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     8221 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     8281 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     8341 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     8401 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     8461 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     8521 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     8581 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     8641 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     8701 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     8761 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     8821 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     8881 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     8941 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     9001 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     9061 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     9121 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     9181 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     9241 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     9301 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     9361 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     9421 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     9481 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     9541 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     9601 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     9661 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     9721 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     9781 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     9841 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     9901 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     9961 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
    10021 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
    10081 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
    10141 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
    10201 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
    10261 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtc
//
