LOCUS       sequence-226692-        8089 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Transcriptional Unit (TU) for YFP expression, obtained by combining 
            FB/GBparts FB007+GB0053+FB008 into pDGB3alpha1R. According to 
            FungalBraid/GoldenBraid modular DNA assembly for ATMT.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 8089)
  AUTHORS   Hernanz-Koers M, Gandia M, Garrigues S, Manzanares P, Yenush L, 
            Orzaez D, Marcos JF
  TITLE     FungalBraid: A GoldenBraid-based modular cloning platform for the 
            assembly and exchange of DNA elements tailored to fungal synthetic 
            biology.
  JOURNAL   Fungal Genet Biol. 2018 Jul;116:51-61. doi: 
            10.1016/j.fgb.2018.04.010. Epub 2018 Apr 20.
  PUBMED    29680684
REFERENCE   2  (bases 1 to 8089)
  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 
            Genet Biol."; date: "2018-07"; volume: "116:51-61. doi"; pages: " 
            10.1016/j.fgb.2018.04.010. Epub 2018 Apr 20"
COMMENT     Sequence Label: FB026
FEATURES             Location/Qualifiers
     source          1..8089
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(join(265..975,976..978,979..981))
                     /codon_start=1
                     /product="enhanced variant of YFP (Heikal et al., 2001)"
                     /label=Citrine
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLKCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(265..975,976..978,979..981))
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLKCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(307..328)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     656..675
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     915..936
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     misc_feature    1731..1755
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             3056..3685
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIGTHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             4119..5186
                     /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      5252..5446
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(5682..5704)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    5790..5930
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     5845..5864
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5945..5962)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6116..6704)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6196..6215)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6792..7607)
                     /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(6792..7607)
                     /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     7515..7534
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     misc_feature    8040..8064
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
ORIGIN
        1 catgtctcag acggtcgatg cattgcagat gagctgtatc tggaagaggt aaacccgaaa
       61 cgcgttttat tcttgttgac atggagctat taaatcacta gaaggcactc tttgctgctt
      121 ggacaaatga acgtatctta tcgagatcct gaacaccatt tgtctcaact ccggagctga
      181 catcgacacc aacgatctta tatccagatt cgtcaagctg tttgatgatt tcagtaacgt
      241 taagtaagct cagctcatga ctgacttgta gagctcgtcc atgccgagag tgatcccggc
      301 ggcggtcacg aactccagca ggaccatgtg atcgcgcttc tcgttggggt ctttgctcag
      361 ggcggactgg tagctcaggt agtggttgtc gggcagcagc acggggccgt cgccgatggg
      421 ggtgttctgc tggtagtggt cggcgagctg cacgctgccg tcctcgatgt tgtggcggat
      481 cttgaagttc accttgatgc cgttcttctg cttgtcggcc atgatataga cgttgtggct
      541 gttgtagttg tactccagct tgtgccccag gatgttgccg tcctccttga agtcgatgcc
      601 cttcagctcg atgcggttca ccagggtgtc gccctcgaac ttcacctcgg cgcgggtctt
      661 gtagttgccg tcgtccttga agaagatggt gcgctcctgg acgtagcctt cgggcatggc
      721 ggacttgaag aagtcgtgct gcttcatgtg gtcggggtag cgggcgaagc acttcaggcc
      781 gtagccgaag gtggtcacga gggtgggcca gggcacgggc agcttgccgg tggtgcagat
      841 gaacttcagg gtcagcttgc cgtaggtggc gtcgccctcg ccctcgccgg acacgctgaa
      901 cttgtggccg tttacgtcgc cgtccagctc gaccaggatg ggcaccaccc cggtgaacag
      961 ctcctcgccc ttgctcacca ttttaaaggt tcttggatgg gaagatgaat atactgaaga
     1021 tgggaaaaga aagagaaaag aaaagagcag ctggtgggga gagcaggaaa atatggcaac
     1081 aaatgttgga ctgacgcaac gaccttgtca accccgccga cacaccgggc ggacagacgg
     1141 ggcaaagctg cctaccaggg actgagggac ctcagcaggt cgagtgcaga gcaccggatg
     1201 ggtcgactgc cagcttgtgt tcccggtctg cgccgctggc cagctcctga gcggcctttc
     1261 cggtttcata caccgggcaa agcaggagag gcacgatatt tggacgccct acagatgccg
     1321 gatgggccaa ttagggagct tacgcgccgg gtactcgctc tacctacttc ggagaaggta
     1381 ctatctcgtg aatcttttac cagatcggaa gcaattggac ttctgtacct aggttaatgg
     1441 catgctattt cgccgacggc tatacacccc tggcttcaca ttctccttcg cttactgccg
     1501 gtgattcgat gaagctccat attctccgat gatgcaatag attcttggtc aacgaggggc
     1561 acaccagcct ttccacttcg gggcggaggg gcggccggtc ccggattaat aatcatccac
     1621 tgcacctcag agccgccaga gctgtctggc gcagtggcgc ttattactca gcccttctct
     1681 ctgcgtccgt ccgtctgtcc gcatgcgcca ctccgtcatg agacggatcc tgacaggata
     1741 tattggcggg taaacctaag agaaaagagc gtttattaga ataatcggat atttaaaagg
     1801 gcgtgaaaag gtttatccgt tcgtccattt gtatgtgcat gccaaccaca gggttcccct
     1861 cgggatcaaa gtactttgat ccaacccctc cgctgctata gtgcagtcgg cttctgacgt
     1921 tcagtgcagc cgtcatctga aaacgacatg tcgcacaagt cctaagttac gcgacaggct
     1981 gccgccctgc ccttttcctg gcgttttctt gtcgcgtgtt ttagtcgcat aaagtagaat
     2041 acttgcgact agaaccggag acattacgcc atgaacaaga gcgccgccgc tggcctgctg
     2101 ggctatgccc gcgtcagcac cgacgaccag gacttgacca accaacgggc cgaactgcac
     2161 gcggccggct gcaccaagct gttttccgag aagatcaccg gcaccaggcg cgaccgcccg
     2221 gagctggcca ggatgcttga ccacctacgc cctggcgacg ttgtgacagt gaccaggcta
     2281 gaccgcctgg cccgcagcac ccgcgaccta ctggacattg ccgagcgcat ccaggaggcc
     2341 ggcgcgggcc tgcgtagcct ggcagagccg tgggccgaca ccaccacgcc ggccggccgc
     2401 atggtgttga ccgtgttcgc cggcattgcc gagttcgagc gttccctaat catcgaccgc
     2461 acccggagcg ggcgcgaggc cgccaaggcc cgaggcgtga agtttggccc ccgccctacc
     2521 ctcaccccgg cacagatcgc gcacgcccgc gagctgatcg accaggaagg ccgcaccgtg
     2581 aaagaggcgg ctgcactgct tggcgtgcat cgctcgaccc tgtaccgcgc acttgagcgc
     2641 agcgaggaag tgacgcccac cgaggccagg cggcgcggtg ccttccgtga ggacgcattg
     2701 accgaggccg acgccctggc ggccgccgag aatgaacgcc aagaggaaca agcatgaaac
     2761 cgcaccagga cggccaggac gaaccgtttt tcattaccga agagatcgag gcggagatga
     2821 tcgcggccgg gtacgtgttc gagccgcccg cgcacctctc aaccgtgcgg ctgcatgaaa
     2881 tcctggccgg tttgtctgat gccaagctgg cggcctggcc ggccagcttg gccgctgaag
     2941 aaaccgagcg ccgccgtcta aaaaggtgat gtgtatttga gtaaaacagc ttgcgtcatg
     3001 cggtcgctgc gtatatgatc cgatgagtaa ataaacaaat acgcaagggg aacgcatgaa
     3061 ggttatcgct gtacttaacc agaaaggcgg gtcaggcaag acgaccatcg gaacccatct
     3121 agcccgcgcc ctgcaactcg ccggggccga tgttctgtta gtcgattccg atccccaggg
     3181 cagtgcccgc gattgggcgg ccgtgcggga agatcaaccg ctaaccgttg tcggcatcga
     3241 ccgcccgacg attgaccgcg acgtgaaggc catcggccgg cgcgacttcg tagtgatcga
     3301 cggagcgccc caggcggcgg acttggctgt gtccgcgatc aaggcagccg acttcgtgct
     3361 gattccggtg cagccaagcc cttacgacat atgggccacc gccgacctgg tggagctggt
     3421 taagcagcgc attgaggtca cggatggaag gctacaagcg gcctttgtcg tgtcgcgggc
     3481 gatcaaaggc acgcgcatcg gcggtgaggt tgccgaggcg ctggccgggt acgagctgcc
     3541 cattcttgag tcccgtatca cgcagcgcgt gagctaccca ggcactgccg ccgccggcac
     3601 aaccgttctt gaatcagaac ccgagggcga cgctgcccgc gaggtccagg cgctggccgc
     3661 tgaaattaaa tcaaaactca tttgagttaa tgaggtaaag agaaaatgag caaaagcaca
     3721 aacacgctaa gtgccggccg tccgagcgca cgcagcagca aggctgcaac gttggccagc
     3781 ctggcagaca cgccagccat gaagcgggtc aactttcagt tgccggcgga ggatcacacc
     3841 aagctgaaga tgtacgcggt acgccaaggc aagaccatta ccgagctgct atctgaatag
     3901 atcgcgcagc taccagagta aatgagcaaa tgaataaatg agtagatgaa ttttagcggc
     3961 taaaggaggc ggcatggaaa atcaagaaca accaggcacc gacgccgtgg aatgccccat
     4021 gtgtggagga acgggcggtt ggccaggcgt aagcggctgg gttgtctgcc ggccctgcaa
     4081 tggcactgga acccccaagc ccgaggaatc ggcgtgacgg tcgcaaacca tccggcccgg
     4141 tacaaatcgg cgcggcgctg ggtgatgacc tggtggagaa gttgaaggcc gcgcaggccg
     4201 cccagcggca acgcatcgag gcagaagcac gccccggtga atcgtggcaa gcggccgctg
     4261 atcgaatccg caaagaatcc cggcaaccgc cggcagccgg tgcgccgtcg attaggaagc
     4321 cgcccaaggg cgacgagcaa ccagattttt tcgttccgat gctctatgac gtgggcaccc
     4381 gcgatagtcg cagcatcatg gacgtggccg ttttccgtct gtcgaagcgt gaccgacgag
     4441 ctggcgaggt gatccgctac gagcttccag acgggcacgt agaggtttcc gcagggccgg
     4501 ccggcatggc cagtgtgtgg gattacgacc tggtactgat ggcggtttcc catctaaccg
     4561 aatccatgaa ccgataccgg gaagggaagg gagacaagcc cggccgcgtg ttccgtccac
     4621 acgttgcgga cgtactcaag ttctgccggc gagccgatgg cggaaagcag aaagacgacc
     4681 tggtagaaac ctgcattcgg ttaaacacca cgcacgttgc catgcagcgt acgaagaagg
     4741 ccaagaacgg ccgcctggtg acggtatccg agggtgaagc cttgattagc cgctacaaga
     4801 tcgtaaagag cgaaaccggg cggccggagt acatcgagat cgagctagct gattggatgt
     4861 accgcgagat cacagaaggc aagaacccgg acgtgctgac ggttcacccc gattactttt
     4921 tgatcgatcc cggcatcggc cgttttctct accgcctggc acgccgcgcc gcaggcaagg
     4981 cagaagccag atggttgttc aagacgatct acgaacgcag tggcagcgcc ggagagttca
     5041 agaagttctg tttcaccgtg cgcaagctga tcgggtcaaa tgacctgccg gagtacgatt
     5101 tgaaggagga ggcggggcag gctggcccga tcctagtcat gcgctaccgc aacctgatcg
     5161 agggcgaagc atccgccggt tcctaatgta cggagcagat gctagggcaa attgccctag
     5221 caggggaaaa aggtcgaaaa ggactctttc ctgtggatag cacgtacatt gggaacccaa
     5281 agccgtacat tgggaaccgg aacccgtaca ttgggaaccc aaagccgtac attgggaacc
     5341 ggtcacacat gtaagtgact gatataaaag agaaaaaagg cgatttttcc gcctaaaact
     5401 ctttaaaact tattaaaact cttaaaaccc gcctggcctg tgcataactg tctggccagc
     5461 gcacagccga agagctgcaa aaagcgccta cccttcggtc gctgcgctcc ctacgccccg
     5521 ccgcttcgcg tcggcctatc gcggccgctg gccgctcaaa aatggctggc ctacggccag
     5581 gcaatctacc agggcgcgga caagccgcgc cgtcgccact cgaccgccgg cgcccacatc
     5641 aaggcaccct gcctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc
     5701 ccggtgacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc
     5761 gcgtcagcgg gtgttggcgg gtgtcggggc gcagccatga cccagtcacg tagcgatagc
     5821 ggagtgtata ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata
     5881 tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgctcttccg
     5941 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     6001 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     6061 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     6121 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     6181 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     6241 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     6301 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     6361 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     6421 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     6481 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     6541 acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     6601 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     6661 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     6721 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatgc
     6781 attctaggtg attagaaaaa ctcatcgagc atcaaatgaa actgcaattt attcatatca
     6841 ggattatcaa taccatattt ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg
     6901 aggcagttcc ataggatggc aagatcctgg tatcggtctg cgattccgac tcgtccaaca
     6961 tcaatacaac ctattaattt cccctcgtca aaaataaggt tatcaagtga gaaatcacca
     7021 tgagtgacga ctgaatccgg tgagaatggc aaaagtttat gcatttcttt ccagacttgt
     7081 tcaacaggcc agccattacg ctcgtcatca aaatcactcg catcaaccaa accgttattc
     7141 attcgtgatt gcgcctgagc gagtcgaaat acgcgatcgc tgttaaaagg acaattacaa
     7201 acaggaatcg aatgcaaccg gcgcaggaac actgccagcg catcaacaat attttcacct
     7261 gaatcaggat attcttctaa tacctggaat gctgttttcc ctgggatcgc agtggtgagt
     7321 aaccatgcat catcaggagt acggataaaa tgcttgatgg tcggaagagg cataaattcc
     7381 gtcagccagt ttagtctgac catctcatct gtaacatcat tggcaacgct acctttgcca
     7441 tgtttcagaa acaactctgg cgcatcgggc ttcccataca atcggtagat tgtcgcacct
     7501 gattgcccga cattatcgcg agcccattta tacccatata aatcagcatc catgttggaa
     7561 tttaatcgcg gccttgagca agacgtttcc cgttgaatat ggctcataac agaacttatt
     7621 atttccttcc tcttttctac agtatttaaa gataccccaa gaagctaatt ataacaagac
     7681 gaactccaat tcactgttcc ttgcattcta aaaccttaaa taccagaaaa cagctttttc
     7741 aaagttgttt tcaaagttgg cgtataacat agtatcgacg gagccgattt tgaaaccgcg
     7801 gtgatcacag gcagcaacgc tctgtcatcg ttacaatcaa catgctaccc tccgcgagat
     7861 catccgtgtt tcaaacccgg cagcttagtt gccgttcttc cgaatagcat cggtaacatg
     7921 agcaaagtct gccgccttac aacggctctc ccgctgacgc cgtcccggac tgatgggctg
     7981 cctgtatcga gtggtgattt tgtgccgagc tgccggtcgg ggagctgttg gctggctggt
     8041 ggcaggatat attgtggtgt aaacataacg gatccgtctc aggagagcg
//
