LOCUS       sequence-381464-        6251 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression vector for fungal (Aspergillus flavus) genes based on 
            pyrithiamine resistance (ptrA) selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6251)
  AUTHORS   Chang PK, Scharfenstein LL, Mack B, Ehrlich KC
  TITLE     Deletion of the Aspergillus flavus orthologue of A. nidulans fluG 
            reduces conidiation and promotes production of sclerotia but does 
            not abolish aflatoxin biosynthesis.
  JOURNAL   Appl Environ Microbiol. 2012 Nov;78(21):7557-63. doi: 
            10.1128/AEM.01241-12. Epub 2012 Aug 17.
  PUBMED    22904054
REFERENCE   2  (bases 1 to 6251)
  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"; doi: 
            "10.1128/AEM.01241-12"; journalName: "Appl Environ Microbiol"; date:
            "2012-11"; volume: "78"; issue: "21"; pages: "7557-63"
COMMENT     Sequence Label: pTR1-GPD-TRPC
FEATURES             Location/Qualifiers
     source          1..6251
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     21..40
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     gene            276..2283
                     /product="pyrithiamine resistance gene of Aspergillus 
                     oryzae (Kubodera et al., 2000)"
                     /label=ptrA
                     /note="useful as a dominant selectable marker in 
                     Aspergillus"
     CDS             complement(589..600)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             join(998..1334,1393..2039)
                     /codon_start=1
                     /gene="ptrA of Aspergillus oryzae"
                     /product="pyrithiamine resistance protein (Kubodera et al.,
                     2000)"
                     /label=ptrA
                     /note="useful as a dominant selectable marker in 
                     Aspergillus"
                     /translation="MSPPAAIYEPTVAATGLKGKVVVSETVPVEGASQTKLLDHFGGKW
                     DEFKFAPIRESQVSRAMTRRYFEDLDKYAESDVVIVGAGSCGLSTAYVLAKARPDLKIA
                     IVEASVSPGGGAWLGGQLFSAMVMRRPAEVFLNELGVPYEEDANPNYVVVKHASLFTST
                     LMSKVLSFPNVKLFNATAVEDLITRPTENGNPQIAGVVVNWTLVTLHHDDHSCMDPNTI
                     NAPVIISTTGHDGPFGAFCAKRLVSMGSVDKLGGMRGLDMSSAEDAIVKNTREVTKGLI
                     IGGMELSEIDGFNRMGPTFGAMVLSGVKAAEEALKVFDERQRECAE"
     rep_origin      complement(2292..2720)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2402..2423)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2614..2633
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2918..2940
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2932..2949
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2933..2949
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     intron          3252..3367
                     /gene="gpdA"
                     /label=gpdA intron
                     /note="intron from the Aspergillus nidulans 
                     glyceraldehyde-3-phosphate dehydrogenase gene (Punt et al.,
                     1990)"
     primer_bind     complement(4488..4504)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4488..4504)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4501..4523)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4512..4528
                     /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(4536..4542,4543..4560,4561..4566))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4581..4602
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4719..4736)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4890..5478)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4970..4989)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5649..6251,1..189,190..258))
                     /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"
ORIGIN
        1 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
       61 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
      121 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
      181 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
      241 gaaatgttga atactcatac tcttcctttt tcaatgggca attgattacg ggatcccatt
      301 ggtaacgaaa tgtaaaagct aggagatcgt ccgccgatgt caggatgatt tcacttgttt
      361 cttgtccggc tcaccggtca aagctaaaga ggagcaaaag gaacggatag aatcgggtgc
      421 cgctgatcta tacggtatag tgcccttatc acgttgactc aacccatgct atttaactca
      481 acccctcctt ctgaacccca ccatcttctt ccttttcctc tcatcccaca caattctcta
      541 tctcagattt gaattccaaa agtcctcgga cgaaactgaa caagtcttcc tcccttcgat
      601 aaacctttgg tgattggaat aactgaccat cttctatagt tcccaaacca accgacaatg
      661 taaatacact cctcgattag ccctctagag ggcatacgat ggaagtcatg gaatactttt
      721 ggctggactc tcacaatgat caaggtatct taggtaacgt ctttggcgtg ggccggtgtt
      781 cgttcccagt catcgatgca ttcacatgcc ctccctaagc tgggccctag actctaggat
      841 cctagtctag aaggacatgg catcgatgga ctgggttcgt tctgagatta tacggctaaa
      901 acttgatctg gataatacca gcgaaaaggg tcatgccttc tctcgttctt cctgttgatg
      961 gaatggctaa cagatgatag tcattgcaac ttgaaacatg tctcctccag ctgccatcta
     1021 cgaacccact gtggccgcta ccggcctcaa gggtaaggtc gtggtttctg agaccgtccc
     1081 cgttgaggga gcttctcaga ccaagctgtt ggaccatttc ggtggcaagt gggacgagtt
     1141 caagttcgcc cctatccgcg aaagccaggt ctctcgtgcc atgaccagac gttactttga
     1201 ggacctggac aagtacgctg aaagtgacgt tgtcattgtt ggtgctggtt cctgcggtct
     1261 gagcactgcg tacgtcttgg ccaaggctcg tccggacctg aagattgcta tcgtcgaggc
     1321 cagcgtctct cctggtcagt agtccatgat ggattgcctt gcactcagct ttccggaact
     1381 aacgtgcaat aggtggcggt gcctggttgg gtggccaact cttttctgct atggtcatgc
     1441 gccgtcccgc ggaagtcttc ctgaacgagc tgggtgttcc ttacgaagag gacgcaaacc
     1501 ccaactacgt tgtcgtcaag cacgcctccc tgtttacctc gacactcatg tcgaaggttc
     1561 tctccttccc caatgtcaag ctcttcaatg ctaccgctgt tgaggacttg atcacccgtc
     1621 cgaccgagaa cggcaacccc cagattgctg gtgttgtcgt caactggacg ctggtcaccc
     1681 ttcaccacga tgatcactcc tgcatggacc ccaacactat caacgctcct gtcatcatca
     1741 gtaccactgg tcacgatggg ccattcggcg ccttctgtgc gaagcgcttg gtgtccatgg
     1801 gcagcgtcga caagctaggt ggcatgcgtg gtctcgacat gagctcggcc gaggatgcca
     1861 tcgtcaagaa cacccgcgag gttactaagg gcttgataat cggcggtatg gagctgtctg
     1921 aaattgatgg ctttaaccgc atgggcccta ccttcggtgc catggttctc agtggtgtca
     1981 aggctgccga ggaggcattg aaggtgttcg acgagcgtca gcgcgagtgt gctgagtaaa
     2041 tgactcacta cccgaatggg ttcagtgcat gaaccggatt tgtcttacgg tctttgacga
     2101 taggggaatg atgattatgt gatagttctg agatttgaat gaactcgtta gctcgtaatc
     2161 cacatgcata tgtaaatggc tgtgtcccgt atgtaacggt ggggcattct agaataatta
     2221 tgtgtaacaa gaaagacagt ataatacaaa caaagatgca agagcggctc atcgtcaccc
     2281 cattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaatagac
     2341 cgaaatcggc aaaatccctt ataaatcaaa agaatagccc gagatagagt tgagtgttgt
     2401 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
     2461 aaccgtctat cagggcgatg gcccactacg tgaaccatca cccaaatcaa gttttttggg
     2521 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
     2581 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
     2641 taaggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
     2701 tgcgccgcta cagggcgcgt actatggttg ctttgacgta tgcggtgtga aataccgcac
     2761 agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc cattcaggct gcgcaactgt
     2821 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt
     2881 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg
     2941 acggccagtg aattcgagct cggtaccgtt actaggttgc agtcaatgca ttaatgcatt
     3001 gcagatgagc tgtatctgga agaggtaaac ccgaaacgcg ttttattctt gttgacatgg
     3061 agctattaaa tcactagaag gcactctttg ctgcttggac aaatgaacgt atcttatcga
     3121 gatcctgaac accatttgtc tcaactccgg agctgacatc gacaccaacg atcttatatc
     3181 cagattcgtc aagctgtttg atgatttcag taacgttaag tggatccggt ccggaattcc
     3241 cggggcggcc gctgctcaag cggggtagct gttagtcaag ctgcgatgaa gtgggaaagc
     3301 tcgaactgaa aggttcaaag gaataaggga tgggaaggat ggagtatgga tgtagcaaag
     3361 tacttactta ggggaaataa aggttcttgg atgggaagat gaatatactg aagatgggaa
     3421 aagaaagaga aaagaaaaga gcagctggtg gggagagcag gaaaatatgg caacaaatgt
     3481 tggactgacg caacgacctt gtcaaccccg ccgacacacc gggcggacag acggggcaaa
     3541 gctgcctacc agggactgag ggacctcagc aggtcgagtg cagagcaccg gatgggtcga
     3601 ctgccagctt gtgttcccgg tctgcgccgc tggccagctc ctgagcggcc tttccggttt
     3661 catacaccgg gcaaagcagg agaggcacga tatttggacg ccctacagat gccggatggg
     3721 ccaattaggg agcttacgcg ccgggtactc gctctaccta cttcggagaa ggtactatct
     3781 cgtgaatctt ttaccagatc ggaagcaatt ggacttctgt acctaggtta atggcatgct
     3841 atttcgccga cggctataca cccctggctt cacattctcc ttcgcttact gccggtgatt
     3901 cgatgaagct ccatattctc cgatgatgca atagattctt ggtcaacgag gggcacacca
     3961 gcctttccac ttcggggcgg aggggcggcc ggtcccggat taataatcat ccactgcacc
     4021 tcagagccgc cagagctgtc tggcgcagtg gcgcttatta ctcagccctt ctctctgcgt
     4081 ccgtccgtct ctccgcatgc cagaaagagt caccggtcac tgtacagagc tcacgagttc
     4141 gtcacatttt tctacaaatg gtggaggcgg cggattttag gctcaagtca tgaccctctg
     4201 ggtcactcca gaatcagcta ggtcaacgaa taaggatgat tctataggaa gatccaggca
     4261 ccggtcaacc atgatctgga cagatttggg agctcggtat aagctctcca ccatcttatt
     4321 ctgtatagtt taggcttaaa gtttatccag gagatgttgc tgaagtcgat ttgagtccac
     4381 ttcctcactg gtagctatac gactttgatg gtcgttgtag gggctgtatt aggtctcgat
     4441 caaacacaaa tagaattaaa tggtactcga agcttggcgt aatcatggtc atagctgttt
     4501 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     4561 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     4621 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     4681 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     4741 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     4801 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     4861 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     4921 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     4981 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     5041 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     5101 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     5161 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     5221 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     5281 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     5341 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     5401 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     5461 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     5521 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     5581 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     5641 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     5701 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     5761 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     5821 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     5881 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     5941 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     6001 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     6061 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     6121 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     6181 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     6241 ccgagttgct c
//
