LOCUS       sequence-89710-e        7780 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For fungal transformation via ATMT. Transformants to be selected by 
            bialaphos resistance..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7780)
  AUTHORS   Sweigard JA, Chumley F, Carroll A, Farrall L, Valent B
  TITLE     A series of vectors for fungal transformation
  JOURNAL   Fungal Genetics Reports, 1997. Vol. 44, Article 19
REFERENCE   2  (bases 1 to 7780)
  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: "Fungal 
            Genetics Reports, 1997. Vol. 44, Article 19"
COMMENT     Sequence Label: pFGL822-8
FEATURES             Location/Qualifiers
     source          1..7780
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..45)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(29..45)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             complement(52..603)
                     /codon_start=1
                     /gene="Streptomyces hygroscopicus bar"
                     /product="phosphinothricin acetyltransferase"
                     /label=BlpR
                     /note="confers resistance to bialophos or phosphinothricin"
                     /translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
                     WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
                     TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
                     WQLDFSLPVPPRPVLPVTEI"
     primer_bind     complement(1006..1023)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1006..1022)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1015..1037)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     misc_feature    1225..1249
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             2550..3179
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             3613..4680
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="GRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESWQA
                     AADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLSKR
                     DRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKPGR
                     VFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGEAL
                     ISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYRLA
                     RRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPILV
                     MRYRNLIEGEASAGS"
     rep_origin      4746..4940
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(5176..5198)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    5284..5424
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     5339..5358
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5439..5456)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5610..6198)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5690..5709)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6285..7079)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     misc_feature    7504..7528
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     polyA_signal    7606..7780
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
ORIGIN
        1 ctcgagaatt cgagctcggt acccggggga tccactagtt ctagagtcga cctaaatctc
       61 ggtgacgggc aggaccggac ggggcggaac cggcaggctg aagtccagct gccagaaacc
      121 cacgtcatgc cagttcccgt gcttgaagcc ggccgcccgc agcatgccgc ggggggcata
      181 tccgagcgcc tcgtgcatgc gcacgctcgg gtcgttgggc agcccgatga cagcgaccac
      241 gctcttgaag ccctgtgcct ccagggactt cagcaggtgg gtgtagagcg tggagcccag
      301 tcccgtccgc tggtggcggg gggagacgta cacggtcgat tcggccgtcc agtcgtaggc
      361 gttgcgtgcc ttccagggac ccgcgtaggc gatgccggcg acctcgccgt ccacctcggc
      421 gacgagccag ggatagcgct cccgcagacg gacgaggtcg tccgtccact cctgcggttc
      481 ctgcggctcg gtacggaagt tgaccgtgct tgtctcgatg tagtggttga cgatggtgca
      541 gaccgccggc atgtccgcct cggtggcacg gcggatgtcg gccgggcgtc gttctgggct
      601 catggttact tcctaatcga agcgttgctt gggtagaata ggtaagtcag attgaatctg
      661 aaataaaggg aggaagggcg aacttaagaa ggtatgaccg ggtcgttcac ttaccttgct
      721 tgacaaacgc accaagttat cgtgcaccaa gcagcagatg ataataatgt cctcgttcct
      781 gtctgctaat aagagtcaca cttcgagcgc cgccgctact gctacaagtg gggctgatct
      841 gaccagttgc ctaaatgaac catcttgtca aacgacacaa attttgtgct caccgcctgg
      901 acgactaaac caaaataggc attcattgtt gacctccact agctccagcc aagcccaaaa
      961 agtgctcctt caatatcact cgacctgcag gcatgcaagc ttggcactgg ccgtcgtttt
     1021 acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc
     1081 ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
     1141 gcgcagcctg aatggcgaat gctagagcag cttgagcttg gatcagattg tcgtttcccg
     1201 ccttcagttt aaactatcag tgtttgacag gatatattgg cgggtaaacc taagagaaaa
     1261 gagcgtttat tagaataatc ggatatttaa aagggcgtga aaaggtttat ccgttggtcc
     1321 atttgtatgt ccatgccaac cacagggttc ccctcgggat caaagtactt tgatccaacc
     1381 cctccgctgc tatagtgcag tcggcttctg acgttcagtg cagccgtctt ctgaaaacga
     1441 catgtcgcac aagtcctaag ttacgcgaca ggctgccgcc ctgccctttt cctggcgttt
     1501 tcttgtcgcg tgttttagtc gcataaagta gaatacttgc gactagaacc ggagacatta
     1561 cgccatgaac aagagcgccg ccgctggcct gctgggctat gcccgcgtca gcaccgacga
     1621 ccaggacttg accaaccaac gggccgaact gcacgcggcc ggctgcacca agctgttttc
     1681 cgagaagatc accggcacca ggcgcgaccg cccggagctg gccaggatgc ttgaccacct
     1741 acgccctggc gacgttgtga cagtgaccag gctagaccgc ctggcccgca gcacccgcga
     1801 cctactggac attgccgagc gcatccagga ggccggcgcg ggcctgcgta gcctggcaga
     1861 gccgtgggcc gacaccacca cgccggccgg ccgcatggtg ttgaccgtgt tcgccggcat
     1921 tgccgagttc gagcgttccc taatcatcga ccgcacccgg agcgggcgcg aggccgccaa
     1981 ggcccgaggc gtgaagtttg gcccccgccc taccctcacc ccggcacaga tcgcgcacgc
     2041 ccgcgagctg atcgaccagg aaggccgcac cgtgaaagag gcggctgcac tgcttggcgt
     2101 gcatcgctcg accctgtacc gcgcacttga gcgcagcgag gaagtgacgc ccaccgaggc
     2161 caggcggcgc ggtgccttcc gtgaggacgc attgaccgag gccgacgccc tggcggccgc
     2221 cgagaatgaa cgccaagagg aacaagcatg aaaccgcacc aggacggcca ggacgaaccg
     2281 tttttcatta ccgaagagat cgaggcggag atgatcgcgg ccgggtacgt gttcgagccg
     2341 cccgcgcacg tctcaaccgt gcggctgcat gaaatcctgg ccggtttgtc tgatgccaag
     2401 ctggcggcct ggccggccag cttggccgct gaagaaaccg agcgccgccg tctaaaaagg
     2461 tgatgtgtat ttgagtaaaa cagcttgcgt catgcggtcg ctgcgtatat gatgcgatga
     2521 gtaaataaac aaatacgcaa ggggaacgca tgaaggttat cgctgtactt aaccagaaag
     2581 gcgggtcagg caagacgacc atcgcaaccc atctagcccg cgccctgcaa ctcgccgggg
     2641 ccgatgttct gttagtcgat tccgatcccc agggcagtgc ccgcgattgg gcggccgtgc
     2701 gggaagatca accgctaacc gttgtcggca tcgaccgccc gacgattgac cgcgacgtga
     2761 aggccatcgg ccggcgcgac ttcgtagtga tcgacggagc gccccaggcg gcggacttgg
     2821 ctgtgtccgc gatcaaggca gccgacttcg tgctgattcc ggtgcagcca agcccttacg
     2881 acatatgggc caccgccgac ctggtggagc tggttaagca gcgcattgag gtcacggatg
     2941 gaaggctaca agcggccttt gtcgtgtcgc gggcgatcaa aggcacgcgc atcggcggtg
     3001 aggttgccga ggcgctggcc gggtacgagc tgcccattct tgagtcccgt atcacgcagc
     3061 gcgtgagcta cccaggcact gccgccgccg gcacaaccgt tcttgaatca gaacccgagg
     3121 gcgacgctgc ccgcgaggtc caggcgctgg ccgctgaaat taaatcaaaa ctcatttgag
     3181 ttaatgaggt aaagagaaaa tgagcaaaag cacaaacacg ctaagtgccg gccgtccgag
     3241 cgcacgcagc agcaaggctg caacgttggc cagcctggca gacacgccag ccatgaagcg
     3301 ggtcaacttt cagttgccgg cggaggatca caccaagctg aagatgtacg cggtacgcca
     3361 aggcaagacc attaccgagc tgctatctga atacatcgcg cagctaccag agtaaatgag
     3421 caaatgaata aatgagtaga tgaattttag cggctaaagg aggcggcatg gaaaatcaag
     3481 aacaaccagg caccgacgcc gtggaatgcc ccatgtgtgg aggaacgggc ggttggccag
     3541 gcgtaagcgg ctgggttgtc tgccggccct gcaatggcac tggaaccccc aagcccgagg
     3601 aatcggcgtg acggtcgcaa accatccggc ccggtacaaa tcggcgcggc gctgggtgat
     3661 gacctggtgg agaagttgaa ggccgcgcag gccgcccagc ggcaacgcat cgaggcagaa
     3721 gcacgccccg gtgaatcgtg gcaagcggcc gctgatcgaa tccgcaaaga atcccggcaa
     3781 ccgccggcag ccggtgcgcc gtcgattagg aagccgccca agggcgacga gcaaccagat
     3841 tttttcgttc cgatgctcta tgacgtgggc acccgcgata gtcgcagcat catggacgtg
     3901 gccgttttcc gtctgtcgaa gcgtgaccga cgagctggcg aggtgatccg ctacgagctt
     3961 ccagacgggc acgtagaggt ttccgcaggg ccggccggca tggccagtgt gtgggattac
     4021 gacctggtac tgatggcggt ttcccatcta accgaatcca tgaaccgata ccgggaaggg
     4081 aagggagaca agcccggccg cgtgttccgt ccacacgttg cggacgtact caagttctgc
     4141 cggcgagccg atggcggaaa gcagaaagac gacctggtag aaacctgcat tcggttaaac
     4201 accacgcacg ttgccatgca gcgtacgaag aaggccaaga acggccgcct ggtgacggta
     4261 tccgagggtg aagccttgat tagccgctac aagatcgtaa agagcgaaac cgggcggccg
     4321 gagtacatcg agatcgagct agctgattgg atgtaccgcg agatcacaga aggcaagaac
     4381 ccggacgtgc tgacggttca ccccgattac tttttgatcg atcccggcat cggccgtttt
     4441 ctctaccgcc tggcacgccg cgccgcaggc aaggcagaag ccagatggtt gttcaagacg
     4501 atctacgaac gcagtggcag cgccggagag ttcaagaagt tctgtttcac cgtgcgcaag
     4561 ctgatcgggt caaatgacct gccggagtac gatttgaagg aggaggcggg gcaggctggc
     4621 ccgatcctag tcatgcgcta ccgcaacctg atcgagggcg aagcatccgc cggttcctaa
     4681 tgtacggagc agatgctagg gcaaattgcc ctagcagggg aaaaaggtcg aaaaggtctc
     4741 tttcctgtgg atagcacgta cattgggaac ccaaagccgt acattgggaa ccggaacccg
     4801 tacattggga acccaaagcc gtacattggg aaccggtcac acatgtaagt gactgatata
     4861 aaagagaaaa aaggcgattt ttccgcctaa aactctttaa aacttattaa aactcttaaa
     4921 acccgcctgg cctgtgcata actgtctggc cagcgcacag ccgaagagct gcaaaaagcg
     4981 cctacccttc ggtcgctgcg ctccctacgc cccgccgctt cgcgtcggcc tatcgcggcc
     5041 gctggccgct caaaaatggc tggcctacgg ccaggcaatc taccagggcg cggacaagcc
     5101 gcgccgtcgc cactcgaccg ccggcgccca catcaaggca ccctgcctcg cgcgtttcgg
     5161 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     5221 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     5281 gggcgcagcc atgacccagt cacgtagcga tagcggagtg tatactggct taactatgcg
     5341 gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc
     5401 gtaaggagaa aataccgcat caggcgctct tccgcttcct cgctcactga ctcgctgcgc
     5461 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     5521 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     5581 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     5641 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     5701 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     5761 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     5821 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     5881 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     5941 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     6001 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     6061 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     6121 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     6181 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     6241 aacgaaaact cacgttaagg gattttggtc atgcattcta ggtactaaaa caattcatcc
     6301 agtaaaatat aatattttat tttctcccaa tcaggcttga tccccagtaa gtcaaaaaat
     6361 agctcgacat actgttcttc cccgatatcc tccctgatcg accggacgca gaaggcaatg
     6421 tcataccact tgtccgccct gccgcttctc ccaagatcaa taaagccact tactttgcca
     6481 tctttcacaa agatgttgct gtctcccagg tcgccgtggg aaaagacaag ttcctcttcg
     6541 ggcttttccg tctttaaaaa atcatacagc tcgcgcggat ctttaaatgg agtgtcttct
     6601 tcccagtttt cgcaatccac atcggccaga tcgttattca gtaagtaatc caattcggct
     6661 aagcggctgt ctaagctatt cgtataggga caatccgata tgtcgatgga gtgaaagagc
     6721 ctgatgcact ccgcatacag ctcgataatc ttttcagggc tttgttcatc ttcatactct
     6781 tccgagcaaa ggacgccatc ggcctcactc atgagcagat tgctccagcc atcatgccgt
     6841 tcaaagtgca ggacctttgg aacaggcagc tttccttcca gccatagcat catgtccttt
     6901 tcccgttcca catcataggt ggtcccttta taccggctgt ccgtcatttt taaatatagg
     6961 ttttcatttt ctcccaccag cttatatacc ttagcaggag acattccttc cgtatctttt
     7021 acgcagcggt atttttcgat cagttttttc aattccggtg atattctcat tttagccatt
     7081 tattatttcc ttcctctttt ctacagtatt taaagatacc ccaagaagct aattataaca
     7141 agacgaactc caattcactg ttccttgcat tctaaaacct taaataccag aaaacagctt
     7201 tttcaaagtt gttttcaaag ttggcgtata acatagtatc gacggagccg attttgaaac
     7261 cgcggtgatc acaggcagca acgctctgtc atcgttacaa tcaacatgct accctccgcg
     7321 agatcatccg tgtttcaaac ccggcagctt agttgccgtt cttccgaata gcatcggtaa
     7381 catgagcaaa gtctgccgcc ttacaacggc tctcccgctg acgccgtccc ggactgatgg
     7441 gctgcctgta tcgagtggtg attttgtgcc gagctgccgg tcggggagct gttggctggc
     7501 tggtggcagg atatattgtg gtgtaaacaa attgacgctt agacaactta ataacacatt
     7561 gcggacgttt ttaatgtact gaattaacgc cgaattaatt cgggggatct ggattttagt
     7621 actggatttt ggttttagga attagaaatt ttattgatag aagtatttta caaatacaaa
     7681 tacatactaa gggtttctta tatgctcaac acatgagcga aaccctatag gaaccctaat
     7741 tcccttatct gggaactact cacacattat tatggagaaa
//
