LOCUS       sequence-89706-e        8252 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For fungal transformation via ATMT. Transformants to be selected by 
            hygromicin resistance..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8252)
  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 8252)
  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: pFGL821-8
FEATURES             Location/Qualifiers
     source          1..8252
                     /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(73..1098)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRSQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     K"
     primer_bind     complement(1478..1495)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1478..1494)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1487..1509)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     misc_feature    1697..1721
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             3022..3651
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             4085..5152
                     /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      5218..5412
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(5648..5670)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    5756..5896
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     5811..5830
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5911..5928)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6082..6670)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6162..6181)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6757..7551)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     misc_feature    7976..8000
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     polyA_signal    8078..8252
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
ORIGIN
        1 ctcgagaatt cgagctcggt acccggggga tccactagtt ctagagtcga ctggttccgg
       61 tcggcatcta ctctatttct ttgccctcgg acgagtgctg gggcgtcggt ttccactatc
      121 ggcgagtact tctacacagc catcggtcca gacggccgcg cttctgcggg cgatttgtgt
      181 acgcccgaca gtcccggctc cggatcggac gattgcgtcg catcgaccct gcgcccaagc
      241 tgcatcatcg aaattgccgt caaccaagct ctgatagagt tggtcaagac caatgcggag
      301 catatacgcc cggagtcgtg gcgatcctgc aagctccgga tgcctccgct cgaagtagcg
      361 cgtctgctgc tccatacaag ccaaccacgg cctccagaag aagatgttgg cgacctcgta
      421 ttgggaatcc ccgaacatcg cctcgctcca gtcaatgacc gctgttatgc ggccattgtc
      481 cgtcaggaca ttgttggagc cgaaatccgc gtgcacgagg tgccggactt cggggcagtc
      541 ctcggcccaa agcatcagct catcgagagc ctgcgcgacg gacgcactga cggtgtcgtc
      601 catcacagtt tgccagtgat acacatgggg atcagcaatc gcgcatatga aatcacgcca
      661 tgtagtgtat tgaccgattc cttgcggtcc gaatgggccg aacccgctcg tctggctaag
      721 atcggccgca gcgatcgcat ccatagcctc cgcgaccggt tgtagaacag cgggcagttc
      781 ggtttcaggc aggtcttgca acgtgacacc ctgtgaacgg cgggagatgc aataggtcag
      841 gctctcgcta aactccccaa tgtcaagcac ttccggaatc gggagcgcgg ccgatgcaaa
      901 gtgccgataa acataacgat ctttgtagaa accatcggcg cagctattta cccgcaggac
      961 atatccacgc cctcctacat cgaagctgaa agcacgagat tcttcgccct ccgagagctg
     1021 catcaggtcg gagacgctgt cgaacttttc gatcagaaac ttctcgacag acgtcgcggt
     1081 gagttcaggc tttttcattt ggatgcttgg gtagaatagg taagtcagat tgaatctgaa
     1141 ataaagggag gaagggcgaa cttaagaagg tatgaccggg tcgtccactt accttgcttg
     1201 acaaacgcac caagttatcg tgcaccaagc agcagatgat aataatgtcc tcgttcctgt
     1261 ctgctaataa gagtcacact tcgagcgccg ccgctactgc tacaagtggg gctgatctga
     1321 ccagttgcct aaatgaacca tcttgtcaaa cgacacaaat tttgtgctca ccgcctggac
     1381 gactaaacca aaataggcat tcattgttga cctccactag ctccagccaa gcccaaaaag
     1441 tgctccttca atatcactcg acctgcagaa gcttggcact ggccgtcgtt ttacaacgtc
     1501 gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg
     1561 ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
     1621 tgaatggcga atgctagagc agcttgagct tggatcagat tgtcgtttcc cgccttcagt
     1681 ttaaactatc agtgtttgac aggatatatt ggcgggtaaa cctaagagaa aagagcgttt
     1741 attagaataa tcggatattt aaaagggcgt gaaaaggttt atccgttggt ccatttgtat
     1801 gtccatgcca accacagggt tcccctcggg atcaaagtac tttgatccaa cccctccgct
     1861 gctatagtgc agtcggcttc tgacgttcag tgcagccgtc ttctgaaaac gacatgtcgc
     1921 acaagtccta agttacgcga caggctgccg ccctgccctt ttcctggcgt tttcttgtcg
     1981 cgtgttttag tcgcataaag tagaatactt gcgactagaa ccggagacat tacgccatga
     2041 acaagagcgc cgccgctggc ctgctgggct atgcccgcgt cagcaccgac gaccaggact
     2101 tgaccaacca acgggccgaa ctgcacgcgg ccggctgcac caagctgttt tccgagaaga
     2161 tcaccggcac caggcgcgac cgcccggagc tggccaggat gcttgaccac ctacgccctg
     2221 gcgacgttgt gacagtgacc aggctagacc gcctggcccg cagcacccgc gacctactgg
     2281 acattgccga gcgcatccag gaggccggcg cgggcctgcg tagcctggca gagccgtggg
     2341 ccgacaccac cacgccggcc ggccgcatgg tgttgaccgt gttcgccggc attgccgagt
     2401 tcgagcgttc cctaatcatc gaccgcaccc ggagcgggcg cgaggccgcc aaggcccgag
     2461 gcgtgaagtt tggcccccgc cctaccctca ccccggcaca gatcgcgcac gcccgcgagc
     2521 tgatcgacca ggaaggccgc accgtgaaag aggcggctgc actgcttggc gtgcatcgct
     2581 cgaccctgta ccgcgcactt gagcgcagcg aggaagtgac gcccaccgag gccaggcggc
     2641 gcggtgcctt ccgtgaggac gcattgaccg aggccgacgc cctggcggcc gccgagaatg
     2701 aacgccaaga ggaacaagca tgaaaccgca ccaggacggc caggacgaac cgtttttcat
     2761 taccgaagag atcgaggcgg agatgatcgc ggccgggtac gtgttcgagc cgcccgcgca
     2821 cgtctcaacc gtgcggctgc atgaaatcct ggccggtttg tctgatgcca agctggcggc
     2881 ctggccggcc agcttggccg ctgaagaaac cgagcgccgc cgtctaaaaa ggtgatgtgt
     2941 atttgagtaa aacagcttgc gtcatgcggt cgctgcgtat atgatgcgat gagtaaataa
     3001 acaaatacgc aaggggaacg catgaaggtt atcgctgtac ttaaccagaa aggcgggtca
     3061 ggcaagacga ccatcgcaac ccatctagcc cgcgccctgc aactcgccgg ggccgatgtt
     3121 ctgttagtcg attccgatcc ccagggcagt gcccgcgatt gggcggccgt gcgggaagat
     3181 caaccgctaa ccgttgtcgg catcgaccgc ccgacgattg accgcgacgt gaaggccatc
     3241 ggccggcgcg acttcgtagt gatcgacgga gcgccccagg cggcggactt ggctgtgtcc
     3301 gcgatcaagg cagccgactt cgtgctgatt ccggtgcagc caagccctta cgacatatgg
     3361 gccaccgccg acctggtgga gctggttaag cagcgcattg aggtcacgga tggaaggcta
     3421 caagcggcct ttgtcgtgtc gcgggcgatc aaaggcacgc gcatcggcgg tgaggttgcc
     3481 gaggcgctgg ccgggtacga gctgcccatt cttgagtccc gtatcacgca gcgcgtgagc
     3541 tacccaggca ctgccgccgc cggcacaacc gttcttgaat cagaacccga gggcgacgct
     3601 gcccgcgagg tccaggcgct ggccgctgaa attaaatcaa aactcatttg agttaatgag
     3661 gtaaagagaa aatgagcaaa agcacaaaca cgctaagtgc cggccgtccg agcgcacgca
     3721 gcagcaaggc tgcaacgttg gccagcctgg cagacacgcc agccatgaag cgggtcaact
     3781 ttcagttgcc ggcggaggat cacaccaagc tgaagatgta cgcggtacgc caaggcaaga
     3841 ccattaccga gctgctatct gaatacatcg cgcagctacc agagtaaatg agcaaatgaa
     3901 taaatgagta gatgaatttt agcggctaaa ggaggcggca tggaaaatca agaacaacca
     3961 ggcaccgacg ccgtggaatg ccccatgtgt ggaggaacgg gcggttggcc aggcgtaagc
     4021 ggctgggttg tctgccggcc ctgcaatggc actggaaccc ccaagcccga ggaatcggcg
     4081 tgacggtcgc aaaccatccg gcccggtaca aatcggcgcg gcgctgggtg atgacctggt
     4141 ggagaagttg aaggccgcgc aggccgccca gcggcaacgc atcgaggcag aagcacgccc
     4201 cggtgaatcg tggcaagcgg ccgctgatcg aatccgcaaa gaatcccggc aaccgccggc
     4261 agccggtgcg ccgtcgatta ggaagccgcc caagggcgac gagcaaccag attttttcgt
     4321 tccgatgctc tatgacgtgg gcacccgcga tagtcgcagc atcatggacg tggccgtttt
     4381 ccgtctgtcg aagcgtgacc gacgagctgg cgaggtgatc cgctacgagc ttccagacgg
     4441 gcacgtagag gtttccgcag ggccggccgg catggccagt gtgtgggatt acgacctggt
     4501 actgatggcg gtttcccatc taaccgaatc catgaaccga taccgggaag ggaagggaga
     4561 caagcccggc cgcgtgttcc gtccacacgt tgcggacgta ctcaagttct gccggcgagc
     4621 cgatggcgga aagcagaaag acgacctggt agaaacctgc attcggttaa acaccacgca
     4681 cgttgccatg cagcgtacga agaaggccaa gaacggccgc ctggtgacgg tatccgaggg
     4741 tgaagccttg attagccgct acaagatcgt aaagagcgaa accgggcggc cggagtacat
     4801 cgagatcgag ctagctgatt ggatgtaccg cgagatcaca gaaggcaaga acccggacgt
     4861 gctgacggtt caccccgatt actttttgat cgatcccggc atcggccgtt ttctctaccg
     4921 cctggcacgc cgcgccgcag gcaaggcaga agccagatgg ttgttcaaga cgatctacga
     4981 acgcagtggc agcgccggag agttcaagaa gttctgtttc accgtgcgca agctgatcgg
     5041 gtcaaatgac ctgccggagt acgatttgaa ggaggaggcg gggcaggctg gcccgatcct
     5101 agtcatgcgc taccgcaacc tgatcgaggg cgaagcatcc gccggttcct aatgtacgga
     5161 gcagatgcta gggcaaattg ccctagcagg ggaaaaaggt cgaaaaggtc tctttcctgt
     5221 ggatagcacg tacattggga acccaaagcc gtacattggg aaccggaacc cgtacattgg
     5281 gaacccaaag ccgtacattg ggaaccggtc acacatgtaa gtgactgata taaaagagaa
     5341 aaaaggcgat ttttccgcct aaaactcttt aaaacttatt aaaactctta aaacccgcct
     5401 ggcctgtgca taactgtctg gccagcgcac agccgaagag ctgcaaaaag cgcctaccct
     5461 tcggtcgctg cgctccctac gccccgccgc ttcgcgtcgg cctatcgcgg ccgctggccg
     5521 ctcaaaaatg gctggcctac ggccaggcaa tctaccaggg cgcggacaag ccgcgccgtc
     5581 gccactcgac cgccggcgcc cacatcaagg caccctgcct cgcgcgtttc ggtgatgacg
     5641 gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg
     5701 ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag
     5761 ccatgaccca gtcacgtagc gatagcggag tgtatactgg cttaactatg cggcatcaga
     5821 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag
     5881 aaaataccgc atcaggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     5941 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     6001 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     6061 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     6121 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     6181 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     6241 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     6301 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     6361 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     6421 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     6481 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
     6541 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     6601 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     6661 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     6721 ctcacgttaa gggattttgg tcatgcattc taggtactaa aacaattcat ccagtaaaat
     6781 ataatatttt attttctccc aatcaggctt gatccccagt aagtcaaaaa atagctcgac
     6841 atactgttct tccccgatat cctccctgat cgaccggacg cagaaggcaa tgtcatacca
     6901 cttgtccgcc ctgccgcttc tcccaagatc aataaagcca cttactttgc catctttcac
     6961 aaagatgttg ctgtctccca ggtcgccgtg ggaaaagaca agttcctctt cgggcttttc
     7021 cgtctttaaa aaatcataca gctcgcgcgg atctttaaat ggagtgtctt cttcccagtt
     7081 ttcgcaatcc acatcggcca gatcgttatt cagtaagtaa tccaattcgg ctaagcggct
     7141 gtctaagcta ttcgtatagg gacaatccga tatgtcgatg gagtgaaaga gcctgatgca
     7201 ctccgcatac agctcgataa tcttttcagg gctttgttca tcttcatact cttccgagca
     7261 aaggacgcca tcggcctcac tcatgagcag attgctccag ccatcatgcc gttcaaagtg
     7321 caggaccttt ggaacaggca gctttccttc cagccatagc atcatgtcct tttcccgttc
     7381 cacatcatag gtggtccctt tataccggct gtccgtcatt tttaaatata ggttttcatt
     7441 ttctcccacc agcttatata ccttagcagg agacattcct tccgtatctt ttacgcagcg
     7501 gtatttttcg atcagttttt tcaattccgg tgatattctc attttagcca tttattattt
     7561 ccttcctctt ttctacagta tttaaagata ccccaagaag ctaattataa caagacgaac
     7621 tccaattcac tgttccttgc attctaaaac cttaaatacc agaaaacagc tttttcaaag
     7681 ttgttttcaa agttggcgta taacatagta tcgacggagc cgattttgaa accgcggtga
     7741 tcacaggcag caacgctctg tcatcgttac aatcaacatg ctaccctccg cgagatcatc
     7801 cgtgtttcaa acccggcagc ttagttgccg ttcttccgaa tagcatcggt aacatgagca
     7861 aagtctgccg ccttacaacg gctctcccgc tgacgccgtc ccggactgat gggctgcctg
     7921 tatcgagtgg tgattttgtg ccgagctgcc ggtcggggag ctgttggctg gctggtggca
     7981 ggatatattg tggtgtaaac aaattgacgc ttagacaact taataacaca ttgcggacgt
     8041 ttttaatgta ctgaattaac gccgaattaa ttcgggggat ctggatttta gtactggatt
     8101 ttggttttag gaattagaaa ttttattgat agaagtattt tacaaataca aatacatact
     8161 aagggtttct tatatgctca acacatgagc gaaaccctat aggaacccta attcccttat
     8221 ctgggaacta ctcacacatt attatggaga aa
//
