LOCUS       sequence-89560-e        9665 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For fungal transformation via ATMT. Transformants to be selected by 
            sulfonylurea (specially chlorimuro ethyl) resistance..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9665)
  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 9665)
  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: pFGL820
FEATURES             Location/Qualifiers
     source          1..9665
                     /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(929..940)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2891..2908)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2891..2907)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2900..2922)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     misc_feature    3110..3134
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             4435..5064
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             5498..6565
                     /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      6631..6825
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(7061..7083)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    7169..7309
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     7224..7243
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(7324..7341)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7495..8083)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7575..7594)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(8170..8964)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     misc_feature    9389..9413
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     polyA_signal    9491..9665
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
ORIGIN
        1 ctcgagaatt cgagctcggt acccggggga tccactagtt ctagagtcga ggtgccaacg
       61 ccacagtgcc ccacatctcc cggctggatt caccgttgag cggggattgt gtggagggac
      121 ttggtcgaaa atttgtgaac ctccgatggc ggcaaacatg actccacgat tcacagccca
      181 aaggcctggc cctgcttcaa ttgtcatcgt ctgacaggtc caggaaaaat agttagctgg
      241 aggctcgtta cttttttttc ccgcttcaac ggccaagcaa gcaagctcat cgtctctcgt
      301 tcttttctcc attattgacg actgagaaca gattcgaaat gcttcgtact gttggccgca
      361 aagccctgag gggctcatcc aagggatgtt cacgaaccat ctcgactctc aagcccgcca
      421 cggcaactat tgccaagccc ggcagcagga ccctttcgac gccagcgacg gcaacagcaa
      481 cgtaagtcag aatcacgagc gagcgaacaa acaggcagag ttacaaacgc catcactacg
      541 ccttgtcttt tcgtctctgt tacgtgtcgc cacccccatg cgagctacac gtctgcgcct
      601 gaagtcacaa ggcttgcaat ttcatgacga tgaacaacac tgacacttct gttacagagc
      661 acctcgaact aagcccagcg ccagcttcaa tgctcgccgc gatccccagc ctctggtcaa
      721 ccctcgctca ggtgaggcag acgaatcgta agttgcgcca acaccgctcc tacaccgcca
      781 agacccatac cgccgtcgca tttggacaag cacaagactg atttgaccgt ggcatagatt
      841 cattggcaag acgggaggag agattttcca cgagatgatg ctgaggcaaa acgtcaagca
      901 tatctgtaag cgtctaattc tacaaattcc taccctcgat ttcaaccaca tattcaacca
      961 catattctga cacatgagtc acagtcggtt accctggcgg tgctatcctt cccgtgttcg
     1021 acgcgatcta caactcgaag cacatcgact ttgttctgcc caagcatgag caaggcgccg
     1081 gccacatggc agagggctat gctcgcgctt caggcaaacc cggcgttgtt ctcgtcacct
     1141 ccggccccgg tgccacaaat gtcatcactc ccatggccga cgctcttgcc gacggtacac
     1201 ctctggttgt attctcagga caggttgtta cctctgatat tggaagcgac gccttccagg
     1261 aggccgacgt cataggcatc tcccggtctt gcaccaagtg gaacgtcatg gttaagagcg
     1321 ctgacgagct cccgaggaga attaacgagg cctttgagat tgccaccagt gggcgacctg
     1381 ggcctgtctt ggtcgatctg cccaaggacg tcacggctag tgtgctgagg agggctatcc
     1441 ccaccgagac ctcgattccc tctattagcg cagcagcacg ggctgtccaa gaggcaggcc
     1501 gaaagcagct tgagcactcc atcaaacgcg tagccgatct cgtcaacatt gccaagaagc
     1561 ccgtcatata tgccggccaa ggtgtcattt tgtcggaagg cggcgttgaa cttctcaagg
     1621 cgcttgccga caaggcctcg attcctgtca ccaccactct gcatggtctg ggagcctttg
     1681 acgagctcga cgagaaggca ctgcacatgc ttggtatgca cggttcggct tatgccaaca
     1741 tgtccatgca agaggccgat ttgatcattg cccttggtgg ccgcttcgat gaccgtgtca
     1801 ctggcagcat ccccaaattt gctcctgccg ccaagctagc tgctgctgaa ggacgcggag
     1861 gtattgtcca cttcgagatt atgcccaaga acatcaacaa ggtcgtccaa gcaacagagg
     1921 ccattgaggg cgacgttgct tcgaacttga agctgttgct ccccaagatt gaacaacgat
     1981 ccatgaccga tcgcaaggag tggttcgacc agatcaagga gtggaaggag aagtggcctc
     2041 tgtcacatta tgagagggcc gagcgtagtg gtctcatcaa gcctcagact ctgatcgagg
     2101 agctgagcaa cctgactgct gaccgcaagg acatgaccta catcacaacc ggtgttggcc
     2161 agcaccaaat gtggacagca caacatttca ggtggaggca cccacggtcc atgatcacct
     2221 ctggcggttt gggaaccatg ggatatggtc tgccggcagc gattggcgcc aaggttgcta
     2281 ggccagatgc tttggtcatt gacatcgacg gcgacgcatc gttcaacatg actctgacag
     2341 agctttcgac ggcggcacag ttcaacattg gcgtcaaggt cattgtcttg aacaacgagg
     2401 agcagggaat ggtgacccaa tggcagaact tgttctacga ggaccgctac tcacatacac
     2461 accagcgcaa cccagacttc atgaagctcg ccgatgcaat ggacgttcaa catcgccgtg
     2521 tttcgaagcc tgacgatgtc gttgatgctc tgacgtggct gatcaacacc gacggccccg
     2581 ctctgcttga ggtgatgact gataagaagg ttcctgttct gcccatggtg cccggaggta
     2641 acggcctgca cgagttcatc acgtttgatg ccagtaagta atatatccaa tgcactcctg
     2701 tggcccattt tgagccttgg gtatgattct agttcaactt gctaaccagt gcatcaactc
     2761 gtataggcaa ggataagcaa cggagagagc tgatgcgcgc gaggacgaat ggcctgcacg
     2821 gttaagttga taatgggaat tgattattgc acgggaattg catgtcgacc tgcaggcatg
     2881 caagcttggc actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc
     2941 aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc
     3001 gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatgctag agcagcttga
     3061 gcttggatca gattgtcgtt tcccgccttc agtttaaact atcagtgttt gacaggatat
     3121 attggcgggt aaacctaaga gaaaagagcg tttattagaa taatcggata tttaaaaggg
     3181 cgtgaaaagg tttatccgtt ggtccatttg tatgtccatg ccaaccacag ggttcccctc
     3241 gggatcaaag tactttgatc caacccctcc gctgctatag tgcagtcggc ttctgacgtt
     3301 cagtgcagcc gtcttctgaa aacgacatgt cgcacaagtc ctaagttacg cgacaggctg
     3361 ccgccctgcc cttttcctgg cgttttcttg tcgcgtgttt tagtcgcata aagtagaata
     3421 cttgcgacta gaaccggaga cattacgcca tgaacaagag cgccgccgct ggcctgctgg
     3481 gctatgcccg cgtcagcacc gacgaccagg acttgaccaa ccaacgggcc gaactgcacg
     3541 cggccggctg caccaagctg ttttccgaga agatcaccgg caccaggcgc gaccgcccgg
     3601 agctggccag gatgcttgac cacctacgcc ctggcgacgt tgtgacagtg accaggctag
     3661 accgcctggc ccgcagcacc cgcgacctac tggacattgc cgagcgcatc caggaggccg
     3721 gcgcgggcct gcgtagcctg gcagagccgt gggccgacac caccacgccg gccggccgca
     3781 tggtgttgac cgtgttcgcc ggcattgccg agttcgagcg ttccctaatc atcgaccgca
     3841 cccggagcgg gcgcgaggcc gccaaggccc gaggcgtgaa gtttggcccc cgccctaccc
     3901 tcaccccggc acagatcgcg cacgcccgcg agctgatcga ccaggaaggc cgcaccgtga
     3961 aagaggcggc tgcactgctt ggcgtgcatc gctcgaccct gtaccgcgca cttgagcgca
     4021 gcgaggaagt gacgcccacc gaggccaggc ggcgcggtgc cttccgtgag gacgcattga
     4081 ccgaggccga cgccctggcg gccgccgaga atgaacgcca agaggaacaa gcatgaaacc
     4141 gcaccaggac ggccaggacg aaccgttttt cattaccgaa gagatcgagg cggagatgat
     4201 cgcggccggg tacgtgttcg agccgcccgc gcacgtctca accgtgcggc tgcatgaaat
     4261 cctggccggt ttgtctgatg ccaagctggc ggcctggccg gccagcttgg ccgctgaaga
     4321 aaccgagcgc cgccgtctaa aaaggtgatg tgtatttgag taaaacagct tgcgtcatgc
     4381 ggtcgctgcg tatatgatgc gatgagtaaa taaacaaata cgcaagggga acgcatgaag
     4441 gttatcgctg tacttaacca gaaaggcggg tcaggcaaga cgaccatcgc aacccatcta
     4501 gcccgcgccc tgcaactcgc cggggccgat gttctgttag tcgattccga tccccagggc
     4561 agtgcccgcg attgggcggc cgtgcgggaa gatcaaccgc taaccgttgt cggcatcgac
     4621 cgcccgacga ttgaccgcga cgtgaaggcc atcggccggc gcgacttcgt agtgatcgac
     4681 ggagcgcccc aggcggcgga cttggctgtg tccgcgatca aggcagccga cttcgtgctg
     4741 attccggtgc agccaagccc ttacgacata tgggccaccg ccgacctggt ggagctggtt
     4801 aagcagcgca ttgaggtcac ggatggaagg ctacaagcgg cctttgtcgt gtcgcgggcg
     4861 atcaaaggca cgcgcatcgg cggtgaggtt gccgaggcgc tggccgggta cgagctgccc
     4921 attcttgagt cccgtatcac gcagcgcgtg agctacccag gcactgccgc cgccggcaca
     4981 accgttcttg aatcagaacc cgagggcgac gctgcccgcg aggtccaggc gctggccgct
     5041 gaaattaaat caaaactcat ttgagttaat gaggtaaaga gaaaatgagc aaaagcacaa
     5101 acacgctaag tgccggccgt ccgagcgcac gcagcagcaa ggctgcaacg ttggccagcc
     5161 tggcagacac gccagccatg aagcgggtca actttcagtt gccggcggag gatcacacca
     5221 agctgaagat gtacgcggta cgccaaggca agaccattac cgagctgcta tctgaataca
     5281 tcgcgcagct accagagtaa atgagcaaat gaataaatga gtagatgaat tttagcggct
     5341 aaaggaggcg gcatggaaaa tcaagaacaa ccaggcaccg acgccgtgga atgccccatg
     5401 tgtggaggaa cgggcggttg gccaggcgta agcggctggg ttgtctgccg gccctgcaat
     5461 ggcactggaa cccccaagcc cgaggaatcg gcgtgacggt cgcaaaccat ccggcccggt
     5521 acaaatcggc gcggcgctgg gtgatgacct ggtggagaag ttgaaggccg cgcaggccgc
     5581 ccagcggcaa cgcatcgagg cagaagcacg ccccggtgaa tcgtggcaag cggccgctga
     5641 tcgaatccgc aaagaatccc ggcaaccgcc ggcagccggt gcgccgtcga ttaggaagcc
     5701 gcccaagggc gacgagcaac cagatttttt cgttccgatg ctctatgacg tgggcacccg
     5761 cgatagtcgc agcatcatgg acgtggccgt tttccgtctg tcgaagcgtg accgacgagc
     5821 tggcgaggtg atccgctacg agcttccaga cgggcacgta gaggtttccg cagggccggc
     5881 cggcatggcc agtgtgtggg attacgacct ggtactgatg gcggtttccc atctaaccga
     5941 atccatgaac cgataccggg aagggaaggg agacaagccc ggccgcgtgt tccgtccaca
     6001 cgttgcggac gtactcaagt tctgccggcg agccgatggc ggaaagcaga aagacgacct
     6061 ggtagaaacc tgcattcggt taaacaccac gcacgttgcc atgcagcgta cgaagaaggc
     6121 caagaacggc cgcctggtga cggtatccga gggtgaagcc ttgattagcc gctacaagat
     6181 cgtaaagagc gaaaccgggc ggccggagta catcgagatc gagctagctg attggatgta
     6241 ccgcgagatc acagaaggca agaacccgga cgtgctgacg gttcaccccg attacttttt
     6301 gatcgatccc ggcatcggcc gttttctcta ccgcctggca cgccgcgccg caggcaaggc
     6361 agaagccaga tggttgttca agacgatcta cgaacgcagt ggcagcgccg gagagttcaa
     6421 gaagttctgt ttcaccgtgc gcaagctgat cgggtcaaat gacctgccgg agtacgattt
     6481 gaaggaggag gcggggcagg ctggcccgat cctagtcatg cgctaccgca acctgatcga
     6541 gggcgaagca tccgccggtt cctaatgtac ggagcagatg ctagggcaaa ttgccctagc
     6601 aggggaaaaa ggtcgaaaag gtctctttcc tgtggatagc acgtacattg ggaacccaaa
     6661 gccgtacatt gggaaccgga acccgtacat tgggaaccca aagccgtaca ttgggaaccg
     6721 gtcacacatg taagtgactg atataaaaga gaaaaaaggc gatttttccg cctaaaactc
     6781 tttaaaactt attaaaactc ttaaaacccg cctggcctgt gcataactgt ctggccagcg
     6841 cacagccgaa gagctgcaaa aagcgcctac ccttcggtcg ctgcgctccc tacgccccgc
     6901 cgcttcgcgt cggcctatcg cggccgctgg ccgctcaaaa atggctggcc tacggccagg
     6961 caatctacca gggcgcggac aagccgcgcc gtcgccactc gaccgccggc gcccacatca
     7021 aggcaccctg cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc
     7081 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
     7141 cgtcagcggg tgttggcggg tgtcggggcg cagccatgac ccagtcacgt agcgatagcg
     7201 gagtgtatac tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat
     7261 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gctcttccgc
     7321 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     7381 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     7441 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     7501 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     7561 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     7621 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     7681 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     7741 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     7801 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     7861 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     7921 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     7981 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     8041 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     8101 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgca
     8161 ttctaggtac taaaacaatt catccagtaa aatataatat tttattttct cccaatcagg
     8221 cttgatcccc agtaagtcaa aaaatagctc gacatactgt tcttccccga tatcctccct
     8281 gatcgaccgg acgcagaagg caatgtcata ccacttgtcc gccctgccgc ttctcccaag
     8341 atcaataaag ccacttactt tgccatcttt cacaaagatg ttgctgtctc ccaggtcgcc
     8401 gtgggaaaag acaagttcct cttcgggctt ttccgtcttt aaaaaatcat acagctcgcg
     8461 cggatcttta aatggagtgt cttcttccca gttttcgcaa tccacatcgg ccagatcgtt
     8521 attcagtaag taatccaatt cggctaagcg gctgtctaag ctattcgtat agggacaatc
     8581 cgatatgtcg atggagtgaa agagcctgat gcactccgca tacagctcga taatcttttc
     8641 agggctttgt tcatcttcat actcttccga gcaaaggacg ccatcggcct cactcatgag
     8701 cagattgctc cagccatcat gccgttcaaa gtgcaggacc tttggaacag gcagctttcc
     8761 ttccagccat agcatcatgt ccttttcccg ttccacatca taggtggtcc ctttataccg
     8821 gctgtccgtc atttttaaat ataggttttc attttctccc accagcttat ataccttagc
     8881 aggagacatt ccttccgtat cttttacgca gcggtatttt tcgatcagtt ttttcaattc
     8941 cggtgatatt ctcattttag ccatttatta tttccttcct cttttctaca gtatttaaag
     9001 ataccccaag aagctaatta taacaagacg aactccaatt cactgttcct tgcattctaa
     9061 aaccttaaat accagaaaac agctttttca aagttgtttt caaagttggc gtataacata
     9121 gtatcgacgg agccgatttt gaaaccgcgg tgatcacagg cagcaacgct ctgtcatcgt
     9181 tacaatcaac atgctaccct ccgcgagatc atccgtgttt caaacccggc agcttagttg
     9241 ccgttcttcc gaatagcatc ggtaacatga gcaaagtctg ccgccttaca acggctctcc
     9301 cgctgacgcc gtcccggact gatgggctgc ctgtatcgag tggtgatttt gtgccgagct
     9361 gccggtcggg gagctgttgg ctggctggtg gcaggatata ttgtggtgta aacaaattga
     9421 cgcttagaca acttaataac acattgcgga cgtttttaat gtactgaatt aacgccgaat
     9481 taattcgggg gatctggatt ttagtactgg attttggttt taggaattag aaattttatt
     9541 gatagaagta ttttacaaat acaaatacat actaagggtt tcttatatgc tcaacacatg
     9601 agcgaaaccc tataggaacc ctaattccct tatctgggaa ctactcacac attattatgg
     9661 agaaa
//
