LOCUS       sequence-238439-        8043 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transcriptional Unit (TU) for hygromicin resistance in fungi 
            obtained by combining FB001+GB0211+FB002 into pDGB3alpha2. According
            to FungalBraid/GoldenBraid modular DNA assembly for ATMT.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8043)
  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 8043)
  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: FB003
FEATURES             Location/Qualifiers
     source          1..8043
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..25
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(320..1345)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             complement(320..1345)
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     misc_feature    1734..1758
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             2191..3006
                     /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             2191..3006
                     /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     complement(2264..2283)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3094..3682
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3583..3602
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3836..3853
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3868..4008
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3934..3953)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4094..4116
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     rep_origin      4352..4546
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(4612..5685)
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
     CDS             complement(6114..6743)
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
ORIGIN
        1 gtttacccgc caatatatcc tgtcaagctt cgtctcaagc gtcatcatgc aacatgcatg
       61 tactgtctga tgtattaaga gtataggggt caacaactag agtgtctgac aggtatatct
      121 agcagtttga tttgaccatt cttttattaa aacactgtaa cccgcaacga agaactcctc
      181 cgcttcaagt caagaatgcg catatgtgcg taaaatcgac aagcccggcg gcattccacc
      241 gagcagggta ttgcctttgg cactttacgt gcgcagagga gcctgaatgt tgagtggaat
      301 gatggtggaa tgatgaagct cattcctttg ccctcggacg agtgctgggg cgtcggtttc
      361 cactatcggc gagtacttct acacagccat cggtccagac ggccgcgctt ctgcgggcga
      421 tttgtgtacg cccgacagtc ccggctccgg atcggacgat tgcgtcgcat cgaccctgcg
      481 cccaagctgc atcatcgaaa ttgccgtcaa ccaagctctg atagagttgg tcaagaccaa
      541 tgcggagcat atacgcccgg agtcgtggcg atcctgcaag ctccggatgc ctccgctcga
      601 agtagcgcgt ctgctgctcc atacaagcca accacggcct ccagaagaag atgttggcga
      661 cctcgtattg ggaatccccg aacattgcct cgctccagtc aatgaccgct gttatgcggc
      721 cattgtccgt caggacattg ttggagccga aatccgcgtg cacgaggtgc cggacttcgg
      781 ggcagtcctc ggcccaaagc atcagctcat cgagagcctg cgcgacggac gcactgacgg
      841 tgtcgtccat cacagtttgc cagtgataca catggggatc agcaatcgcg catatgaaat
      901 cacgccatgt agtgtattga ccgattcctt gcggtccgaa tgggccgaac ccgctcgtct
      961 ggctaagatc ggccgcagcg atcgcatcca tggcctccgc gaccggctgc agaacagcgg
     1021 gcagttcggt ttcaggcagg tcttgcaacg tgacaccctg tgcacggcgg gagatgcaat
     1081 aggtcaggct ctcgctgaat tccccaatgt caagcacttc cggaatcggg agcgcggccg
     1141 atgcaaagtg ccgataaaca taacgatctt tgtagaaacc atcggcgcag ctatttaccc
     1201 gcaggacata tccacgccct cctacatcga agctgaaagc acgagattct tcgccctccg
     1261 agagctgcat caggtcggac acgctgtcga acttttcgat cagaaacttc tcgacagacg
     1321 tcgcggtgag ttcaggcttt ttcattttgg atgcttgggt agaataggta agtcagattg
     1381 aatctgaaat aaagggagga agggcgaact taagaaggta tgaccgggtc gtccacttac
     1441 cttgcttgac aaacgcacca agttatcgtg caccaagcag cagatgataa taatgtcctc
     1501 gttcctgtct gctaataaga gtcacacttc gagcgccgcc gctactgcta caagtggggc
     1561 tgatctgacc agttgcctaa atgaaccatc ttgtcaaacg acacaaattt tgtgctcacc
     1621 gcctggacga ctaaaccaaa ataggcattc attgttgacc tccactagct ccagccaagc
     1681 ccaaaaaatg ctccttcaat atcagttctc ctgactgaga cgaagcttgt tatgtttaca
     1741 ccacaatata tcctgccacc agccagccaa cagctccccg accggcagct cggcacaaaa
     1801 tcaccactcg atacaggcag cccatcagtc cgggacggcg tcagcgggag agccgttgta
     1861 aggcggcaga ctttgctcat gttaccgatg ctattcggaa gaacggcaac taagctgccg
     1921 ggtttgaaac acggatgatc tcgcggaggg tagcatgttg attgtaacga tgacagagcg
     1981 ttgctgcctg tgatcaccgc ggtttcaaaa tcggctccgt cgatactatg ttatacgcca
     2041 actttgaaaa caactttgaa aaagctgttt tctggtattt aaggttttag aatgcaagga
     2101 acagtgaatt ggagttcgtc ttgttataat tagcttcttg gggtatcttt aaatactgta
     2161 gaaaagagga aggaaataat aagttctgtt atgagccata ttcaacggga aacgtcttgc
     2221 tcaaggccgc gattaaattc caacatggat gctgatttat atgggtataa atgggctcgc
     2281 gataatgtcg ggcaatcagg tgcgacaatc taccgattgt atgggaagcc cgatgcgcca
     2341 gagttgtttc tgaaacatgg caaaggtagc gttgccaatg atgttacaga tgagatggtc
     2401 agactaaact ggctgacgga atttatgcct cttccgacca tcaagcattt tatccgtact
     2461 cctgatgatg catggttact caccactgcg atcccaggga aaacagcatt ccaggtatta
     2521 gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg
     2581 ttgcattcga ttcctgtttg taattgtcct tttaacagcg atcgcgtatt tcgactcgct
     2641 caggcgcaat cacgaatgaa taacggtttg gttgatgcga gtgattttga tgacgagcgt
     2701 aatggctggc ctgttgaaca agtctggaaa gaaatgcata aacttttgcc attctcaccg
     2761 gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga cgaggggaaa
     2821 ttaataggtt gtattgatgt tggacgagtc ggaatcgcag accgatacca ggatcttgcc
     2881 atcctatgga actgcctcgg tgagttttct ccttcattac agaaacggct ttttcaaaaa
     2941 tatggtattg ataatcctga tatgaataaa ttgcagtttc atttgatgct cgatgagttt
     3001 ttctaatcac ctagaatgca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     3061 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     3121 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     3181 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     3241 ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     3301 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     3361 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     3421 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     3481 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     3541 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     3601 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     3661 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     3721 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     3781 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     3841 gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg
     3901 cggtatttca caccgcatat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     3961 aagccagtat acactccgct atcgctacgt gactgggtca tggctgcgcc ccgacacccg
     4021 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
     4081 gctgtgaccg tcaccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
     4141 gcgaggcagg gtgccttgat gtgggcgccg gcggtcgagt ggcgacggcg cggcttgtcc
     4201 gcgccctggt agattgcctg gccgtaggcc agccattttt gagcggccag cggccgcgat
     4261 aggccgacgc gaagcggcgg ggcgtaggga gcgcagcgac cgaagggtag gcgctttttg
     4321 cagctcttcg gctgtgcgct ggccagacag ttatgcacag gccaggcggg ttttaagagt
     4381 tttaataagt tttaaagagt tttaggcgga aaaatcgcct tttttctctt ttatatcagt
     4441 cacttacatg tgtgaccggt tcccaatgta cggctttggg ttcccaatgt acgggttccg
     4501 gttcccaatg tacggctttg ggttcccaat gtacgtgcta tccacaggaa agagtccttt
     4561 tcgacctttt tcccctgcta gggcaatttg ccctagcatc tgctccgtac attaggaacc
     4621 ggcggatgct tcgccctcga tcaggttgcg gtagcgcatg actaggatcg ggccagcctg
     4681 ccccgcctcc tccttcaaat cgtactccgg caggtcattt gacccgatca gcttgcgcac
     4741 ggtgaaacag aacttcttga actctccggc gctgccactg cgttcgtaga tcgtcttgaa
     4801 caaccatctg gcttctgcct tgcctgcggc gcggcgtgcc aggcggtaga gaaaacggcc
     4861 gatgccggga tcgatcaaaa agtaatcggg gtgaaccgtc agcacgtccg ggttcttgcc
     4921 ttctgtgatc tcgcggtaca tccaatcagc tagctcgatc tcgatgtact ccggccgccc
     4981 ggtttcgctc tttacgatct tgtagcggct aatcaaggct tcaccctcgg ataccgtcac
     5041 caggcggccg ttcttggcct tcttcgtacg ctgcatggca acgtgcgtgg tgtttaaccg
     5101 aatgcaggtt tctaccaggt cgtctttctg ctttccgcca tcggctcgcc ggcagaactt
     5161 gagtacgtcc gcaacgtgtg gacggaacac gcggccgggc ttgtctccct tcccttcccg
     5221 gtatcggttc atggattcgg ttagatggga aaccgccatc agtaccaggt cgtaatccca
     5281 cacactggcc atgccggccg gccctgcgga aacctctacg tgcccgtctg gaagctcgta
     5341 gcggatcacc tcgccagctc gtcggtcacg cttcgacaga cggaaaacgg ccacgtccat
     5401 gatgctgcga ctatcgcggg tgcccacgtc atagagcatc ggaacgaaaa aatctggttg
     5461 ctcgtcgccc ttgggcggct tcctaatcga cggcgcaccg gctgccggcg gttgccggga
     5521 ttctttgcgg attcgatcag cggccgcttg ccacgattca ccggggcgtg cttctgcctc
     5581 gatgcgttgc cgctgggcgg cctgcgcggc cttcaacttc tccaccaggt catcacccag
     5641 cgccgcgccg atttgtaccg ggccggatgg tttgcgaccg ctcacgccga ttcctcgggc
     5701 ttgggggttc cagtgccatt gcagggccgg caggcaaccc agccgcttac gcctggccaa
     5761 ccgcccgttc ctccacacat ggggcattcc acggcgtcgg tgcctggttg ttcttgattt
     5821 tccatgccgc ctcctttagc cgctaaaatt catctactca tttattcatt tgctcattta
     5881 ctctggtagc tgcgcgatgt attcagatag cagctcggta atggtcttgc cttggcgtac
     5941 cgcgtacatc ttcagcttgg tgtgatcctc cgccggcaac tgaaagttga cccgcttcat
     6001 ggctggcgtg tctgccaggc tggccaacgt tgcagccttg ctgctgcgtg cgctcggacg
     6061 gccggcactt agcgtgtttg tgcttttgct cattttctct ttacctcatt aactcaaatg
     6121 agttttgatt taatttcagc ggccagcgcc tggacctcgc gggcagcgtc gccctcgggt
     6181 tctgattcaa gaacggttgt gccggcggcg gcagtgcctg ggtagctcac gcgctgcgtg
     6241 atacgggact caagaatggg cagctcgtac ccggccagcg cctcggcaac ctcaccgccg
     6301 atgcgcgtgc ctttgatcgc ccgcgacacg acaaaggccg cttgtagcct tccatccgtg
     6361 acctcaatgc gctgcttaac cagctccacc aggtcggcgg tggcccatat gtcgtaaggg
     6421 cttggctgca ccggaatcag cacgaagtcg gctgccttga tcgcggacac agccaagtcc
     6481 gccgcctggg gcgctccgtc gatcactacg aagtcgcgcc ggccgatggc cttcacgtcg
     6541 cggtcaatcg tcgggcggtc gatgccgaca acggttagcg gttgatcttc ccgcacggcc
     6601 gcccaatcgc gggcactgcc ctggggatcg gaatcgacta acagaacatc ggccccggcg
     6661 agttgcaggg cgcgggctag atgggttgcg atggtcgtct tgcctgaccc gcctttctgg
     6721 ttaagtacag cgataacctt catgcgttcc ccttgcgtat ttgtttattt actcatcgca
     6781 tcatatacgc agcgaccgca tgacgcaagc tgttttactc aaatacacat caccttttta
     6841 gacggcggcg ctcggtttct tcagcggcca agctggccgg ccaggccgcc agcttggcat
     6901 cagacaaacc ggccaggatt tcatgcagcc gcacggttga gaggtgcgcg ggcggctcga
     6961 acacgtaccc ggccgcgatc atctccgcct cgatctcttc ggtaatgaaa aacggttcgt
     7021 cctggccgtc ctggtgcggt ttcatgcttg ttcctcttgg cgttcattct cggcggccgc
     7081 cagggcgtcg gcctcggtca atgcgtcctc acggaaggca ccgcgccgcc tggcctcggt
     7141 gggcgtcact tcctcgctgc gctcaagtgc gcggtacagg gtcgagcgat gcacgccaag
     7201 cagtgcagcc gcctctttca cggtgcggcc ttcctggtcg atcagctcgc gggcgtgcgc
     7261 gatctgtgcc ggggtgaggg tagggcgggg gccaaacttc acgcctcggg ccttggcggc
     7321 ctcgcgcccg ctccgggtgc ggtcgatgat tagggaacgc tcgaactcgg caatgccggc
     7381 gaacacggtc aacaccatgc ggccggccgg cgtggtggtg tcggcccacg gctctgccag
     7441 gctacgcagg cccgcgccgg cctcctggat gcgctcggca atgtccagta ggtcgcgggt
     7501 gctgcgggcc aggcggtcta gcctggtcac tgtcacaacg tcgccagggc gtaggtggtc
     7561 aagcatcctg gccagctccg ggcggtcgcg cctggtgccg gtgatcttct cggaaaacag
     7621 cttggtgcag ccggccgcgt gcagttcggc ccgttggttg gtcaagtcct ggtcgtcggt
     7681 gctgacgcgg gcatagccca gcaggccagc ggcggcgctc ttgttcatgg cgtaatgtct
     7741 ccggttctag tcgcaagtat tctactttat gcgactaaaa cacgcgacaa gaaaacgcca
     7801 ggaaaagggc agggcggcag cctgtcgcgt aacttaggac ttgtgcgaca tgtcgttttc
     7861 agatgacggc tgcactgaac gtcagaagcc gactgcacta tagcagcgga ggggttggat
     7921 caaagtactt tgatcccgag gggaaccctg tggttggcat gcacatacaa atggacgaac
     7981 ggataaacct tttcacgccc ttttaaatat ccgattattc taataaacgc tcttttctct
     8041 tag
//
