LOCUS       sequence-89704-e        8251 bp    DNA     circular SYN 14-APR-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 8251)
  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 8251)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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
FEATURES             Location/Qualifiers
     source          1..8251
                     /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             409..1434
                     /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(1477..1494)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1477..1493)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1486..1508)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     misc_feature    1696..1720
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             3021..3650
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             4084..5151
                     /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      5217..5411
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(5647..5669)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    5755..5895
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     5810..5829
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5910..5927)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6081..6669)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6161..6180)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6756..7550)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     misc_feature    7975..7999
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     polyA_signal    8077..8251
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
ORIGIN
        1 ctcgagaatt cgagctcggt acccggggga tccactagtt ctagagtcga gtatattgaa
       61 ggagcacttt ttgggcttgg ctggagctag tggaggtcaa caatgaatgc ctattttggt
      121 ttagtcgtcc aggcggtgag cacaaaattt gtgtcgtttg acaagatggt tcatttaggc
      181 aactggtcag atcagcccca cttgtagcag tagcggcggc gctcgaagtg tgactcttat
      241 tagcagacag gaacgaggac attattatca tctgctgctt ggtgcacgat aacttggtgc
      301 gtttgtcaag caaggtaagt ggacgacccg gtcatacctt cttaagttcg cccttcctcc
      361 ctttatttca gattcaatct gacttaccta ttctacccaa gcatccaaat gaaaaagcct
      421 gaactcaccg cgacgtctgt cgagaagttt ctgatcgaaa agttcgacag cgtctccgac
      481 ctgatgcagc tctcggaggg cgaagaatct cgtgctttca gcttcgatgt aggagggcgt
      541 ggatatgtcc tgcgggtaaa tagctgcgcc gatggtttct acaaagatcg ttatgtttat
      601 cggcactttg catcggccgc gctcccgatt ccggaagtgc ttgacattgg ggagtttagc
      661 gagagcctga cctattgcat ctcccgccgt tcacagggtg tcacgttgca agacctgcct
      721 gaaaccgaac tgcccgctgt tctacaaccg gtcgcggagg ctatggatgc gatcgctgcg
      781 gccgatctta gccagacgag cgggttcggc ccattcggac cgcaaggaat cggtcaatac
      841 actacatggc gtgatttcat atgcgcgatt gctgatcccc atgtgtatca ctggcaaact
      901 gtgatggacg acaccgtcag tgcgtccgtc gcgcaggctc tcgatgagct gatgctttgg
      961 gccgaggact gccccgaagt ccggcacctc gtgcacgcgg atttcggctc caacaatgtc
     1021 ctgacggaca atggccgcat aacagcggtc attgactgga gcgaggcgat gttcggggat
     1081 tcccaatacg aggtcgccaa catcttcttc tggaggccgt ggttggcttg tatggagcag
     1141 cagacgcgct acttcgagcg gaggcatccg gagcttgcag gatcgccacg actccgggcg
     1201 tatatgctcc gcattggtct tgaccaactc tatcagagct tggttgacgg caatttcgat
     1261 gatgcagctt gggcgcaggg tcgatgcgac gcaatcgtcc gatccggagc cgggactgtc
     1321 gggcgtacac aaatcgcccg cagaagcgcg gccgtctgga ccgatggctg tgtagaagta
     1381 ctcgccgata gtggaaaccg acgccccagc actcgtccga gggcaaagaa atagagtaga
     1441 tgccgaccgg aaccagtcga cctgcagaag cttggcactg gccgtcgttt tacaacgtcg
     1501 tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc
     1561 cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct
     1621 gaatggcgaa tgctagagca gcttgagctt ggatcagatt gtcgtttccc gccttcagtt
     1681 taaactatca gtgtttgaca ggatatattg gcgggtaaac ctaagagaaa agagcgttta
     1741 ttagaataat cggatattta aaagggcgtg aaaaggttta tccgttggtc catttgtatg
     1801 tccatgccaa ccacagggtt cccctcggga tcaaagtact ttgatccaac ccctccgctg
     1861 ctatagtgca gtcggcttct gacgttcagt gcagccgtct tctgaaaacg acatgtcgca
     1921 caagtcctaa gttacgcgac aggctgccgc cctgcccttt tcctggcgtt ttcttgtcgc
     1981 gtgttttagt cgcataaagt agaatacttg cgactagaac cggagacatt acgccatgaa
     2041 caagagcgcc gccgctggcc tgctgggcta tgcccgcgtc agcaccgacg accaggactt
     2101 gaccaaccaa cgggccgaac tgcacgcggc cggctgcacc aagctgtttt ccgagaagat
     2161 caccggcacc aggcgcgacc gcccggagct ggccaggatg cttgaccacc tacgccctgg
     2221 cgacgttgtg acagtgacca ggctagaccg cctggcccgc agcacccgcg acctactgga
     2281 cattgccgag cgcatccagg aggccggcgc gggcctgcgt agcctggcag agccgtgggc
     2341 cgacaccacc acgccggccg gccgcatggt gttgaccgtg ttcgccggca ttgccgagtt
     2401 cgagcgttcc ctaatcatcg accgcacccg gagcgggcgc gaggccgcca aggcccgagg
     2461 cgtgaagttt ggcccccgcc ctaccctcac cccggcacag atcgcgcacg cccgcgagct
     2521 gatcgaccag gaaggccgca ccgtgaaaga ggcggctgca ctgcttggcg tgcatcgctc
     2581 gaccctgtac cgcgcacttg agcgcagcga ggaagtgacg cccaccgagg ccaggcggcg
     2641 cggtgccttc cgtgaggacg cattgaccga ggccgacgcc ctggcggccg ccgagaatga
     2701 acgccaagag gaacaagcat gaaaccgcac caggacggcc aggacgaacc gtttttcatt
     2761 accgaagaga tcgaggcgga gatgatcgcg gccgggtacg tgttcgagcc gcccgcgcac
     2821 gtctcaaccg tgcggctgca tgaaatcctg gccggtttgt ctgatgccaa gctggcggcc
     2881 tggccggcca gcttggccgc tgaagaaacc gagcgccgcc gtctaaaaag gtgatgtgta
     2941 tttgagtaaa acagcttgcg tcatgcggtc gctgcgtata tgatgcgatg agtaaataaa
     3001 caaatacgca aggggaacgc atgaaggtta tcgctgtact taaccagaaa ggcgggtcag
     3061 gcaagacgac catcgcaacc catctagccc gcgccctgca actcgccggg gccgatgttc
     3121 tgttagtcga ttccgatccc cagggcagtg cccgcgattg ggcggccgtg cgggaagatc
     3181 aaccgctaac cgttgtcggc atcgaccgcc cgacgattga ccgcgacgtg aaggccatcg
     3241 gccggcgcga cttcgtagtg atcgacggag cgccccaggc ggcggacttg gctgtgtccg
     3301 cgatcaaggc agccgacttc gtgctgattc cggtgcagcc aagcccttac gacatatggg
     3361 ccaccgccga cctggtggag ctggttaagc agcgcattga ggtcacggat ggaaggctac
     3421 aagcggcctt tgtcgtgtcg cgggcgatca aaggcacgcg catcggcggt gaggttgccg
     3481 aggcgctggc cgggtacgag ctgcccattc ttgagtcccg tatcacgcag cgcgtgagct
     3541 acccaggcac tgccgccgcc ggcacaaccg ttcttgaatc agaacccgag ggcgacgctg
     3601 cccgcgaggt ccaggcgctg gccgctgaaa ttaaatcaaa actcatttga gttaatgagg
     3661 taaagagaaa atgagcaaaa gcacaaacac gctaagtgcc ggccgtccga gcgcacgcag
     3721 cagcaaggct gcaacgttgg ccagcctggc agacacgcca gccatgaagc gggtcaactt
     3781 tcagttgccg gcggaggatc acaccaagct gaagatgtac gcggtacgcc aaggcaagac
     3841 cattaccgag ctgctatctg aatacatcgc gcagctacca gagtaaatga gcaaatgaat
     3901 aaatgagtag atgaatttta gcggctaaag gaggcggcat ggaaaatcaa gaacaaccag
     3961 gcaccgacgc cgtggaatgc cccatgtgtg gaggaacggg cggttggcca ggcgtaagcg
     4021 gctgggttgt ctgccggccc tgcaatggca ctggaacccc caagcccgag gaatcggcgt
     4081 gacggtcgca aaccatccgg cccggtacaa atcggcgcgg cgctgggtga tgacctggtg
     4141 gagaagttga aggccgcgca ggccgcccag cggcaacgca tcgaggcaga agcacgcccc
     4201 ggtgaatcgt ggcaagcggc cgctgatcga atccgcaaag aatcccggca accgccggca
     4261 gccggtgcgc cgtcgattag gaagccgccc aagggcgacg agcaaccaga ttttttcgtt
     4321 ccgatgctct atgacgtggg cacccgcgat agtcgcagca tcatggacgt ggccgttttc
     4381 cgtctgtcga agcgtgaccg acgagctggc gaggtgatcc gctacgagct tccagacggg
     4441 cacgtagagg tttccgcagg gccggccggc atggccagtg tgtgggatta cgacctggta
     4501 ctgatggcgg tttcccatct aaccgaatcc atgaaccgat accgggaagg gaagggagac
     4561 aagcccggcc gcgtgttccg tccacacgtt gcggacgtac tcaagttctg ccggcgagcc
     4621 gatggcggaa agcagaaaga cgacctggta gaaacctgca ttcggttaaa caccacgcac
     4681 gttgccatgc agcgtacgaa gaaggccaag aacggccgcc tggtgacggt atccgagggt
     4741 gaagccttga ttagccgcta caagatcgta aagagcgaaa ccgggcggcc ggagtacatc
     4801 gagatcgagc tagctgattg gatgtaccgc gagatcacag aaggcaagaa cccggacgtg
     4861 ctgacggttc accccgatta ctttttgatc gatcccggca tcggccgttt tctctaccgc
     4921 ctggcacgcc gcgccgcagg caaggcagaa gccagatggt tgttcaagac gatctacgaa
     4981 cgcagtggca gcgccggaga gttcaagaag ttctgtttca ccgtgcgcaa gctgatcggg
     5041 tcaaatgacc tgccggagta cgatttgaag gaggaggcgg ggcaggctgg cccgatccta
     5101 gtcatgcgct accgcaacct gatcgagggc gaagcatccg ccggttccta atgtacggag
     5161 cagatgctag ggcaaattgc cctagcaggg gaaaaaggtc gaaaaggtct ctttcctgtg
     5221 gatagcacgt acattgggaa cccaaagccg tacattggga accggaaccc gtacattggg
     5281 aacccaaagc cgtacattgg gaaccggtca cacatgtaag tgactgatat aaaagagaaa
     5341 aaaggcgatt tttccgccta aaactcttta aaacttatta aaactcttaa aacccgcctg
     5401 gcctgtgcat aactgtctgg ccagcgcaca gccgaagagc tgcaaaaagc gcctaccctt
     5461 cggtcgctgc gctccctacg ccccgccgct tcgcgtcggc ctatcgcggc cgctggccgc
     5521 tcaaaaatgg ctggcctacg gccaggcaat ctaccagggc gcggacaagc cgcgccgtcg
     5581 ccactcgacc gccggcgccc acatcaaggc accctgcctc gcgcgtttcg gtgatgacgg
     5641 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
     5701 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggcgcagc
     5761 catgacccag tcacgtagcg atagcggagt gtatactggc ttaactatgc ggcatcagag
     5821 cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga
     5881 aaataccgca tcaggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     5941 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     6001 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     6061 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     6121 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     6181 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     6241 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     6301 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     6361 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     6421 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     6481 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc
     6541 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     6601 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     6661 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     6721 tcacgttaag ggattttggt catgcattct aggtactaaa acaattcatc cagtaaaata
     6781 taatatttta ttttctccca atcaggcttg atccccagta agtcaaaaaa tagctcgaca
     6841 tactgttctt ccccgatatc ctccctgatc gaccggacgc agaaggcaat gtcataccac
     6901 ttgtccgccc tgccgcttct cccaagatca ataaagccac ttactttgcc atctttcaca
     6961 aagatgttgc tgtctcccag gtcgccgtgg gaaaagacaa gttcctcttc gggcttttcc
     7021 gtctttaaaa aatcatacag ctcgcgcgga tctttaaatg gagtgtcttc ttcccagttt
     7081 tcgcaatcca catcggccag atcgttattc agtaagtaat ccaattcggc taagcggctg
     7141 tctaagctat tcgtataggg acaatccgat atgtcgatgg agtgaaagag cctgatgcac
     7201 tccgcataca gctcgataat cttttcaggg ctttgttcat cttcatactc ttccgagcaa
     7261 aggacgccat cggcctcact catgagcaga ttgctccagc catcatgccg ttcaaagtgc
     7321 aggacctttg gaacaggcag ctttccttcc agccatagca tcatgtcctt ttcccgttcc
     7381 acatcatagg tggtcccttt ataccggctg tccgtcattt ttaaatatag gttttcattt
     7441 tctcccacca gcttatatac cttagcagga gacattcctt ccgtatcttt tacgcagcgg
     7501 tatttttcga tcagtttttt caattccggt gatattctca ttttagccat ttattatttc
     7561 cttcctcttt tctacagtat ttaaagatac cccaagaagc taattataac aagacgaact
     7621 ccaattcact gttccttgca ttctaaaacc ttaaatacca gaaaacagct ttttcaaagt
     7681 tgttttcaaa gttggcgtat aacatagtat cgacggagcc gattttgaaa ccgcggtgat
     7741 cacaggcagc aacgctctgt catcgttaca atcaacatgc taccctccgc gagatcatcc
     7801 gtgtttcaaa cccggcagct tagttgccgt tcttccgaat agcatcggta acatgagcaa
     7861 agtctgccgc cttacaacgg ctctcccgct gacgccgtcc cggactgatg ggctgcctgt
     7921 atcgagtggt gattttgtgc cgagctgccg gtcggggagc tgttggctgg ctggtggcag
     7981 gatatattgt ggtgtaaaca aattgacgct tagacaactt aataacacat tgcggacgtt
     8041 tttaatgtac tgaattaacg ccgaattaat tcgggggatc tggattttag tactggattt
     8101 tggttttagg aattagaaat tttattgata gaagtatttt acaaatacaa atacatacta
     8161 agggtttctt atatgctcaa cacatgagcg aaaccctata ggaaccctaa ttcccttatc
     8221 tgggaactac tcacacatta ttatggagaa a
//
