LOCUS       sequence-403777-        9048 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Assembly of the transcriptional unit for the auxotrophy marker pyr4 
            from T. reseei..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9048)
  AUTHORS   Moreno-Gimenez E, Gandia M, Saez Z, Manzanares P, Yenush L, Orzaez 
            D, Marcos JF, Garrigues S
  TITLE     FungalBraid 2.0: expanding the synthetic biology toolbox for the 
            biotechnological exploitation of filamentous fungi.
  JOURNAL   Front Bioeng Biotechnol. 2023 Aug 7;11:1222812. doi: 
            10.3389/fbioe.2023.1222812. eCollection 2023.
  PUBMED    37609115
REFERENCE   2  (bases 1 to 9048)
  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: "Front 
            Bioeng Biotechnol."; date: "2023-08-7"; volume: "11:1222812. doi"; 
            pages: " 10.3389/fbioe.2023.1222812. eCollection 2023"
COMMENT     Sequence Label: FB293
FEATURES             Location/Qualifiers
     source          1..9048
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    2693..2717
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             4018..4647
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIGTHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             5081..6148
                     /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      6214..6408
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(6644..6666)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    6752..6892
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     6807..6826
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6907..6924)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7078..7666)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7158..7177)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7754..8569)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(7754..8569)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     8477..8496
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     misc_feature    9002..9026
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
ORIGIN
        1 ggaggatctg tgtcttttgc cttgccttgt cttgtctggg ttcctgctcg tctgcggcaa
       61 tcggaacgcc gccagtgcgg tgccaagcaa accagccaag gtaggtacct accactaggc
      121 ttcttttctc gttgtctcac tctctctttt cctctttgtc ctctcttatc cccatctttt
      181 ctctctctct ctgctccttt cctaaccact tccctacctt tctctttttc cttttcttgt
      241 catctccatc ttggctgacg aaaaaggtct gactgggtag gtattatctg gcagacttgt
      301 gtgtatcatt caccctattt ctgcttcata gtacatgtac tgtacctgaa cggctcaacc
      361 gctatttacg actcttattt ttttgtggcg ttggtcacgt ttgccagctg ttgtccgtct
      421 ttctagggct cctcaaactt gacctgaccg agctcccttt ctggacccgg tgggcttcac
      481 ttccagctgc tgagcgacct gagccgaaca tcctcagtcc ttgtccagcg caattcattt
      541 tctttccttt tctttttttt tattcctttc tttactttta ttctctcttt ttctcctctt
      601 cctcttcttc ttctttctcc tcctcctcca tatcctcact ctcgtttccc tcattactac
      661 cctctcggct cctcaggtcc accaaccctc ccgcacccaa acctctgccg ctgaaaccca
      721 ttcggtggtc gccgtttttt tttttttttt tttctcaccc ccaaagtcgc aatatcgggt
      781 atcgccgccg gcattgaatc gccttctccg ctagcatcga ctactgctgc tctgctctcg
      841 ttgccagcgc tgctccctag aattttgacc aggggacgag cccgacatta aagcaactcc
      901 ctcgcctcga gatgactcgg atcgcacgaa attctcccaa tcgccgacag ttcctactcc
      961 tcttcctccc gcacggctgt cgcgcttcca acgtcattcg cacagcagaa ttgtgccatc
     1021 tctctctttt ttttcccccc ctctaaaccg ccacaacggc accctaaggg ttaaactatc
     1081 caaccagccg cagcctcagc ctctctcagc ctcatcagcc aatggcacca cacccgacgc
     1141 tcaaggccac cttcgcggcc aggagcgaga ctgcgacgca cccgctgacg gcttacctgt
     1201 tcaagctcat ggacctcaag gcatccaacc tgtgcctgag cgccgacgtg ccgacagcgc
     1261 gcgagctgct gtacctggcc gacaagattg gtccgtcgat tgtcgtgctc aagacgcact
     1321 acgacatggt ttcgggctgg gacttccacc cggagactgg cacgggagcc cagctggcgt
     1381 cgctggctcg caagcacggc ttcctcatct tcgaggaccg caagtttggt gacattggcc
     1441 acaccgtcga gctgcagtac acgggtgggt cggctcgcat catcgactgg gctcacattg
     1501 tcaacgtcaa catggtgccc ggtaaggcgt cggtggcctc gctggcccag ggtgccaagc
     1561 gctggctcga gcgctacccc tgcgaggtca agacgtccgt caccgtcggc acgcccacca
     1621 tggactcgtt tgacgacgac gccgactcca gggacgccga gccagccggt gccgtcaacg
     1681 gtatgggttc cattggtgtc ctggacaagc ccatctactc gaaccggtcc ggcgacggcc
     1741 gcaagggaag catcgtttcc atcaccaccg tcacccagca gtacgagtcc gtttcctcgc
     1801 cacggttaac aaaggccatc gccgagggtg acgagtcgct cttcccggga atcgaggagg
     1861 ctccgctgag ccgcggcctc ctgatcctcg cccaaatgtc cagccagggc aacttcatga
     1921 acaaggagta cacgcaggcc tgcgtcgagg ccgcccggga gcacaaggac tttgtcatgg
     1981 gcttcatctc gcaggagact ctcaacaccg agcccgacga tgcctttatc cacatgacgc
     2041 ccggctgcca gctgcccccc gaagacgagg accagcagac caacggatcg gtcggtggag
     2101 atggccaggg ccagcagtac aacacgccgc acaagctgat tggcatcgcc ggcagcgaca
     2161 ttgccattgt gggccggggc atcctcaagg cctcagaccc cgtagaggag gcagagcggt
     2221 accgatcagc agcgtggaaa gcctacaccg agaggctgct gcgataggct tgggagggaa
     2281 gggaagaaag aagtaaagaa aggcatttag caagaagggg gaaaagggag ggaggacaaa
     2341 cggagctgag aaagagctct tgtccaaagc ccggcatcat agaatgcagc tgtatttagg
     2401 cgacctcttt ttccatcttg tcgatttttg ttatgacgta ccagttggga tgatggatga
     2461 ttgtacccca gctgcgattg atgtgtatct ttgcatgcaa caacacgcga tggcggaggc
     2521 gaactgcaca ttggaaggtt catatatggt cctgacatat ctggtggatc tggaagcatg
     2581 gaattgtatt tttgatttgg catttgcttt tgcgcgtgga gggaacatat caccctcggg
     2641 catttttcat ttggtaggat ggtttggatg cagttcgctt gagacgaagc tttgacagga
     2701 tatattggcg ggtaaaccta agagaaaaga gcgtttatta gaataatcgg atatttaaaa
     2761 gggcgtgaaa aggtttatcc gttcgtccat ttgtatgtgc atgccaacca cagggttccc
     2821 ctcgggatca aagtactttg atccaacccc tccgctgcta tagtgcagtc ggcttctgac
     2881 gttcagtgca gccgtcatct gaaaacgaca tgtcgcacaa gtcctaagtt acgcgacagg
     2941 ctgccgccct gcccttttcc tggcgttttc ttgtcgcgtg ttttagtcgc ataaagtaga
     3001 atacttgcga ctagaaccgg agacattacg ccatgaacaa gagcgccgcc gctggcctgc
     3061 tgggctatgc ccgcgtcagc accgacgacc aggacttgac caaccaacgg gccgaactgc
     3121 acgcggccgg ctgcaccaag ctgttttccg agaagatcac cggcaccagg cgcgaccgcc
     3181 cggagctggc caggatgctt gaccacctac gccctggcga cgttgtgaca gtgaccaggc
     3241 tagaccgcct ggcccgcagc acccgcgacc tactggacat tgccgagcgc atccaggagg
     3301 ccggcgcggg cctgcgtagc ctggcagagc cgtgggccga caccaccacg ccggccggcc
     3361 gcatggtgtt gaccgtgttc gccggcattg ccgagttcga gcgttcccta atcatcgacc
     3421 gcacccggag cgggcgcgag gccgccaagg cccgaggcgt gaagtttggc ccccgcccta
     3481 ccctcacccc ggcacagatc gcgcacgccc gcgagctgat cgaccaggaa ggccgcaccg
     3541 tgaaagaggc ggctgcactg cttggcgtgc atcgctcgac cctgtaccgc gcacttgagc
     3601 gcagcgagga agtgacgccc accgaggcca ggcggcgcgg tgccttccgt gaggacgcat
     3661 tgaccgaggc cgacgccctg gcggccgccg agaatgaacg ccaagaggaa caagcatgaa
     3721 accgcaccag gacggccagg acgaaccgtt tttcattacc gaagagatcg aggcggagat
     3781 gatcgcggcc gggtacgtgt tcgagccgcc cgcgcacctc tcaaccgtgc ggctgcatga
     3841 aatcctggcc ggtttgtctg atgccaagct ggcggcctgg ccggccagct tggccgctga
     3901 agaaaccgag cgccgccgtc taaaaaggtg atgtgtattt gagtaaaaca gcttgcgtca
     3961 tgcggtcgct gcgtatatga tccgatgagt aaataaacaa atacgcaagg ggaacgcatg
     4021 aaggttatcg ctgtacttaa ccagaaaggc gggtcaggca agacgaccat cggaacccat
     4081 ctagcccgcg ccctgcaact cgccggggcc gatgttctgt tagtcgattc cgatccccag
     4141 ggcagtgccc gcgattgggc ggccgtgcgg gaagatcaac cgctaaccgt tgtcggcatc
     4201 gaccgcccga cgattgaccg cgacgtgaag gccatcggcc ggcgcgactt cgtagtgatc
     4261 gacggagcgc cccaggcggc ggacttggct gtgtccgcga tcaaggcagc cgacttcgtg
     4321 ctgattccgg tgcagccaag cccttacgac atatgggcca ccgccgacct ggtggagctg
     4381 gttaagcagc gcattgaggt cacggatgga aggctacaag cggcctttgt cgtgtcgcgg
     4441 gcgatcaaag gcacgcgcat cggcggtgag gttgccgagg cgctggccgg gtacgagctg
     4501 cccattcttg agtcccgtat cacgcagcgc gtgagctacc caggcactgc cgccgccggc
     4561 acaaccgttc ttgaatcaga acccgagggc gacgctgccc gcgaggtcca ggcgctggcc
     4621 gctgaaatta aatcaaaact catttgagtt aatgaggtaa agagaaaatg agcaaaagca
     4681 caaacacgct aagtgccggc cgtccgagcg cacgcagcag caaggctgca acgttggcca
     4741 gcctggcaga cacgccagcc atgaagcggg tcaactttca gttgccggcg gaggatcaca
     4801 ccaagctgaa gatgtacgcg gtacgccaag gcaagaccat taccgagctg ctatctgaat
     4861 agatcgcgca gctaccagag taaatgagca aatgaataaa tgagtagatg aattttagcg
     4921 gctaaaggag gcggcatgga aaatcaagaa caaccaggca ccgacgccgt ggaatgcccc
     4981 atgtgtggag gaacgggcgg ttggccaggc gtaagcggct gggttgtctg ccggccctgc
     5041 aatggcactg gaacccccaa gcccgaggaa tcggcgtgac ggtcgcaaac catccggccc
     5101 ggtacaaatc ggcgcggcgc tgggtgatga cctggtggag aagttgaagg ccgcgcaggc
     5161 cgcccagcgg caacgcatcg aggcagaagc acgccccggt gaatcgtggc aagcggccgc
     5221 tgatcgaatc cgcaaagaat cccggcaacc gccggcagcc ggtgcgccgt cgattaggaa
     5281 gccgcccaag ggcgacgagc aaccagattt tttcgttccg atgctctatg acgtgggcac
     5341 ccgcgatagt cgcagcatca tggacgtggc cgttttccgt ctgtcgaagc gtgaccgacg
     5401 agctggcgag gtgatccgct acgagcttcc agacgggcac gtagaggttt ccgcagggcc
     5461 ggccggcatg gccagtgtgt gggattacga cctggtactg atggcggttt cccatctaac
     5521 cgaatccatg aaccgatacc gggaagggaa gggagacaag cccggccgcg tgttccgtcc
     5581 acacgttgcg gacgtactca agttctgccg gcgagccgat ggcggaaagc agaaagacga
     5641 cctggtagaa acctgcattc ggttaaacac cacgcacgtt gccatgcagc gtacgaagaa
     5701 ggccaagaac ggccgcctgg tgacggtatc cgagggtgaa gccttgatta gccgctacaa
     5761 gatcgtaaag agcgaaaccg ggcggccgga gtacatcgag atcgagctag ctgattggat
     5821 gtaccgcgag atcacagaag gcaagaaccc ggacgtgctg acggttcacc ccgattactt
     5881 tttgatcgat cccggcatcg gccgttttct ctaccgcctg gcacgccgcg ccgcaggcaa
     5941 ggcagaagcc agatggttgt tcaagacgat ctacgaacgc agtggcagcg ccggagagtt
     6001 caagaagttc tgtttcaccg tgcgcaagct gatcgggtca aatgacctgc cggagtacga
     6061 tttgaaggag gaggcggggc aggctggccc gatcctagtc atgcgctacc gcaacctgat
     6121 cgagggcgaa gcatccgccg gttcctaatg tacggagcag atgctagggc aaattgccct
     6181 agcaggggaa aaaggtcgaa aaggactctt tcctgtggat agcacgtaca ttgggaaccc
     6241 aaagccgtac attgggaacc ggaacccgta cattgggaac ccaaagccgt acattgggaa
     6301 ccggtcacac atgtaagtga ctgatataaa agagaaaaaa ggcgattttt ccgcctaaaa
     6361 ctctttaaaa cttattaaaa ctcttaaaac ccgcctggcc tgtgcataac tgtctggcca
     6421 gcgcacagcc gaagagctgc aaaaagcgcc tacccttcgg tcgctgcgct ccctacgccc
     6481 cgccgcttcg cgtcggccta tcgcggccgc tggccgctca aaaatggctg gcctacggcc
     6541 aggcaatcta ccagggcgcg gacaagccgc gccgtcgcca ctcgaccgcc ggcgcccaca
     6601 tcaaggcacc ctgcctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc
     6661 tcccggtgac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
     6721 gcgcgtcagc gggtgttggc gggtgtcggg gcgcagccat gacccagtca cgtagcgata
     6781 gcggagtgta tactggctta actatgcggc atcagagcag attgtactga gagtgcacca
     6841 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgctcttc
     6901 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     6961 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     7021 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     7081 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     7141 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     7201 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     7261 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     7321 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     7381 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     7441 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     7501 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     7561 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     7621 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     7681 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     7741 gcattctagg tgattagaaa aactcatcga gcatcaaatg aaactgcaat ttattcatat
     7801 caggattatc aataccatat ttttgaaaaa gccgtttctg taatgaagga gaaaactcac
     7861 cgaggcagtt ccataggatg gcaagatcct ggtatcggtc tgcgattccg actcgtccaa
     7921 catcaataca acctattaat ttcccctcgt caaaaataag gttatcaagt gagaaatcac
     7981 catgagtgac gactgaatcc ggtgagaatg gcaaaagttt atgcatttct ttccagactt
     8041 gttcaacagg ccagccatta cgctcgtcat caaaatcact cgcatcaacc aaaccgttat
     8101 tcattcgtga ttgcgcctga gcgagtcgaa atacgcgatc gctgttaaaa ggacaattac
     8161 aaacaggaat cgaatgcaac cggcgcagga acactgccag cgcatcaaca atattttcac
     8221 ctgaatcagg atattcttct aatacctgga atgctgtttt ccctgggatc gcagtggtga
     8281 gtaaccatgc atcatcagga gtacggataa aatgcttgat ggtcggaaga ggcataaatt
     8341 ccgtcagcca gtttagtctg accatctcat ctgtaacatc attggcaacg ctacctttgc
     8401 catgtttcag aaacaactct ggcgcatcgg gcttcccata caatcggtag attgtcgcac
     8461 ctgattgccc gacattatcg cgagcccatt tatacccata taaatcagca tccatgttgg
     8521 aatttaatcg cggccttgag caagacgttt cccgttgaat atggctcata acagaactta
     8581 ttatttcctt cctcttttct acagtattta aagatacccc aagaagctaa ttataacaag
     8641 acgaactcca attcactgtt ccttgcattc taaaacctta aataccagaa aacagctttt
     8701 tcaaagttgt tttcaaagtt ggcgtataac atagtatcga cggagccgat tttgaaaccg
     8761 cggtgatcac aggcagcaac gctctgtcat cgttacaatc aacatgctac cctccgcgag
     8821 atcatccgtg tttcaaaccc ggcagcttag ttgccgttct tccgaatagc atcggtaaca
     8881 tgagcaaagt ctgccgcctt acaacggctc tcccgctgac gccgtcccgg actgatgggc
     8941 tgcctgtatc gagtggtgat tttgtgccga gctgccggtc ggggagctgt tggctggctg
     9001 gtggcaggat atattgtggt gtaaacataa caagcttcgt ctcagtca
//
