LOCUS       sequence-236964-        8042 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transcriptional Unit (TU) for hygromicin resistance in fungi 
            obtained by combining FB001+GB0211+FB002 into pDGB3alpha1. 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 8042)
  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 8042)
  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: FB010
FEATURES             Location/Qualifiers
     source          1..8042
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..25
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             409..1434
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             409..1434
                     /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    1733..1757
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             3058..3687
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             4116..5189
                     /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"
     rep_origin      5255..5449
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(5685..5707)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    5793..5933
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     5848..5867
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5948..5965)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6119..6707)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6199..6218)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6795..7610)
                     /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(6795..7610)
                     /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     7518..7537
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 tggcaggata tattgtggtg taaacataac gaattcgtct caggagaact gatattgaag
       61 gagcattttt tgggcttggc tggagctagt ggaggtcaac aatgaatgcc tattttggtt
      121 tagtcgtcca ggcggtgagc acaaaatttg tgtcgtttga caagatggtt catttaggca
      181 actggtcaga tcagccccac ttgtagcagt agcggcggcg ctcgaagtgt gactcttatt
      241 agcagacagg aacgaggaca ttattatcat ctgctgcttg gtgcacgata acttggtgcg
      301 tttgtcaagc aaggtaagtg gacgacccgg tcataccttc ttaagttcgc ccttcctccc
      361 tttatttcag attcaatctg acttacctat tctacccaag catccaaaat gaaaaagcct
      421 gaactcaccg cgacgtctgt cgagaagttt ctgatcgaaa agttcgacag cgtgtccgac
      481 ctgatgcagc tctcggaggg cgaagaatct cgtgctttca gcttcgatgt aggagggcgt
      541 ggatatgtcc tgcgggtaaa tagctgcgcc gatggtttct acaaagatcg ttatgtttat
      601 cggcactttg catcggccgc gctcccgatt ccggaagtgc ttgacattgg ggaattcagc
      661 gagagcctga cctattgcat ctcccgccgt gcacagggtg tcacgttgca agacctgcct
      721 gaaaccgaac tgcccgctgt tctgcagccg gtcgcggagg ccatggatgc 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 gcgaggcaat 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 gggcaaagga atgagcttca
     1441 tcattccacc atcattccac tcaacattca ggctcctctg cgcacgtaaa gtgccaaagg
     1501 caataccctg ctcggtggaa tgccgccggg cttgtcgatt ttacgcacat atgcgcattc
     1561 ttgacttgaa gcggaggagt tcttcgttgc gggttacagt gttttaataa aagaatggtc
     1621 aaatcaaact gctagatata cctgtcagac actctagttg ttgaccccta tactcttaat
     1681 acatcagaca gtacatgcat gttgcatgat gacgctgtca tgagacgaat tctgacagga
     1741 tatattggcg ggtaaaccta agagaaaaga gcgtttatta gaataatcgg atatttaaaa
     1801 gggcgtgaaa aggtttatcc gttcgtccat ttgtatgtgc atgccaacca cagggttccc
     1861 ctcgggatca aagtactttg atccaacccc tccgctgcta tagtgcagtc ggcttctgac
     1921 gttcagtgca gccgtcatct gaaaacgaca tgtcgcacaa gtcctaagtt acgcgacagg
     1981 ctgccgccct gcccttttcc tggcgttttc ttgtcgcgtg ttttagtcgc ataaagtaga
     2041 atacttgcga ctagaaccgg agacattacg ccatgaacaa gagcgccgcc gctggcctgc
     2101 tgggctatgc ccgcgtcagc accgacgacc aggacttgac caaccaacgg gccgaactgc
     2161 acgcggccgg ctgcaccaag ctgttttccg agaagatcac cggcaccagg cgcgaccgcc
     2221 cggagctggc caggatgctt gaccacctac gccctggcga cgttgtgaca gtgaccaggc
     2281 tagaccgcct ggcccgcagc acccgcgacc tactggacat tgccgagcgc atccaggagg
     2341 ccggcgcggg cctgcgtagc ctggcagagc cgtgggccga caccaccacg ccggccggcc
     2401 gcatggtgtt gaccgtgttc gccggcattg ccgagttcga gcgttcccta atcatcgacc
     2461 gcacccggag cgggcgcgag gccgccaagg cccgaggcgt gaagtttggc ccccgcccta
     2521 ccctcacccc ggcacagatc gcgcacgccc gcgagctgat cgaccaggaa ggccgcaccg
     2581 tgaaagaggc ggctgcactg cttggcgtgc atcgctcgac cctgtaccgc gcacttgagc
     2641 gcagcgagga agtgacgccc accgaggcca ggcggcgcgg tgccttccgt gaggacgcat
     2701 tgaccgaggc cgacgccctg gcggccgccg agaatgaacg ccaagaggaa caagcatgaa
     2761 accgcaccag gacggccagg acgaaccgtt tttcattacc gaagagatcg aggcggagat
     2821 gatcgcggcc gggtacgtgt tcgagccgcc cgcgcacctc tcaaccgtgc ggctgcatga
     2881 aatcctggcc ggtttgtctg atgccaagct ggcggcctgg ccggccagct tggccgctga
     2941 agaaaccgag cgccgccgtc taaaaaggtg atgtgtattt gagtaaaaca gcttgcgtca
     3001 tgcggtcgct gcgtatatga tgcgatgagt aaataaacaa atacgcaagg ggaacgcatg
     3061 aaggttatcg ctgtacttaa ccagaaaggc gggtcaggca agacgaccat cgcaacccat
     3121 ctagcccgcg ccctgcaact cgccggggcc gatgttctgt tagtcgattc cgatccccag
     3181 ggcagtgccc gcgattgggc ggccgtgcgg gaagatcaac cgctaaccgt tgtcggcatc
     3241 gaccgcccga cgattgaccg cgacgtgaag gccatcggcc ggcgcgactt cgtagtgatc
     3301 gacggagcgc cccaggcggc ggacttggct gtgtccgcga tcaaggcagc cgacttcgtg
     3361 ctgattccgg tgcagccaag cccttacgac atatgggcca ccgccgacct ggtggagctg
     3421 gttaagcagc gcattgaggt cacggatgga aggctacaag cggcctttgt cgtgtcgcgg
     3481 gcgatcaaag gcacgcgcat cggcggtgag gttgccgagg cgctggccgg gtacgagctg
     3541 cccattcttg agtcccgtat cacgcagcgc gtgagctacc caggcactgc cgccgccggc
     3601 acaaccgttc ttgaatcaga acccgagggc gacgctgccc gcgaggtcca ggcgctggcc
     3661 gctgaaatta aatcaaaact catttgagtt aatgaggtaa agagaaaatg agcaaaagca
     3721 caaacacgct aagtgccggc cgtccgagcg cacgcagcag caaggctgca acgttggcca
     3781 gcctggcaga cacgccagcc atgaagcggg tcaactttca gttgccggcg gaggatcaca
     3841 ccaagctgaa gatgtacgcg gtacgccaag gcaagaccat taccgagctg ctatctgaat
     3901 acatcgcgca gctaccagag taaatgagca aatgaataaa tgagtagatg aattttagcg
     3961 gctaaaggag gcggcatgga aaatcaagaa caaccaggca ccgacgccgt ggaatgcccc
     4021 atgtgtggag gaacgggcgg ttggccaggc gtaagcggct gggttgcctg ccggccctgc
     4081 aatggcactg gaacccccaa gcccgaggaa tcggcgtgag cggtcgcaaa ccatccggcc
     4141 cggtacaaat cggcgcggcg ctgggtgatg acctggtgga gaagttgaag gccgcgcagg
     4201 ccgcccagcg gcaacgcatc gaggcagaag cacgccccgg tgaatcgtgg caagcggccg
     4261 ctgatcgaat ccgcaaagaa tcccggcaac cgccggcagc cggtgcgccg tcgattagga
     4321 agccgcccaa gggcgacgag caaccagatt ttttcgttcc gatgctctat gacgtgggca
     4381 cccgcgatag tcgcagcatc atggacgtgg ccgttttccg tctgtcgaag cgtgaccgac
     4441 gagctggcga ggtgatccgc tacgagcttc cagacgggca cgtagaggtt tccgcagggc
     4501 cggccggcat ggccagtgtg tgggattacg acctggtact gatggcggtt tcccatctaa
     4561 ccgaatccat gaaccgatac cgggaaggga agggagacaa gcccggccgc gtgttccgtc
     4621 cacacgttgc ggacgtactc aagttctgcc ggcgagccga tggcggaaag cagaaagacg
     4681 acctggtaga aacctgcatt cggttaaaca ccacgcacgt tgccatgcag cgtacgaaga
     4741 aggccaagaa cggccgcctg gtgacggtat ccgagggtga agccttgatt agccgctaca
     4801 agatcgtaaa gagcgaaacc gggcggccgg agtacatcga gatcgagcta gctgattgga
     4861 tgtaccgcga gatcacagaa ggcaagaacc cggacgtgct gacggttcac cccgattact
     4921 ttttgatcga tcccggcatc ggccgttttc tctaccgcct ggcacgccgc gccgcaggca
     4981 aggcagaagc cagatggttg ttcaagacga tctacgaacg cagtggcagc gccggagagt
     5041 tcaagaagtt ctgtttcacc gtgcgcaagc tgatcgggtc aaatgacctg ccggagtacg
     5101 atttgaagga ggaggcgggg caggctggcc cgatcctagt catgcgctac cgcaacctga
     5161 tcgagggcga agcatccgcc ggttcctaat gtacggagca gatgctaggg caaattgccc
     5221 tagcagggga aaaaggtcga aaaggactct ttcctgtgga tagcacgtac attgggaacc
     5281 caaagccgta cattgggaac cggaacccgt acattgggaa cccaaagccg tacattggga
     5341 accggtcaca catgtaagtg actgatataa aagagaaaaa aggcgatttt tccgcctaaa
     5401 actctttaaa acttattaaa actcttaaaa cccgcctggc ctgtgcataa ctgtctggcc
     5461 agcgcacagc cgaagagctg caaaaagcgc ctacccttcg gtcgctgcgc tccctacgcc
     5521 ccgccgcttc gcgtcggcct atcgcggccg ctggccgctc aaaaatggct ggcctacggc
     5581 caggcaatct accagggcgc ggacaagccg cgccgtcgcc actcgaccgc cggcgcccac
     5641 atcaaggcac cctgcctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag
     5701 ctcccggtga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag
     5761 ggcgcgtcag cgggtgttgg cgggtgtcgg ggcgcagcca tgacccagtc acgtagcgat
     5821 agcggagtgt atactggctt aactatgcgg catcagagca gattgtactg agagtgcacc
     5881 atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgctctt
     5941 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     6001 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     6061 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     6121 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     6181 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     6241 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     6301 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     6361 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     6421 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     6481 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     6541 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
     6601 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     6661 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     6721 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     6781 tgcattctag gtgattagaa aaactcatcg agcatcaaat gaaactgcaa tttattcata
     6841 tcaggattat caataccata tttttgaaaa agccgtttct gtaatgaagg agaaaactca
     6901 ccgaggcagt tccataggat ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca
     6961 acatcaatac aacctattaa tttcccctcg tcaaaaataa ggttatcaag tgagaaatca
     7021 ccatgagtga cgactgaatc cggtgagaat ggcaaaagtt tatgcatttc tttccagact
     7081 tgttcaacag gccagccatt acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta
     7141 ttcattcgtg attgcgcctg agcgagtcga aatacgcgat cgctgttaaa aggacaatta
     7201 caaacaggaa tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac aatattttca
     7261 cctgaatcag gatattcttc taatacctgg aatgctgttt tccctgggat cgcagtggtg
     7321 agtaaccatg catcatcagg agtacggata aaatgcttga tggtcggaag aggcataaat
     7381 tccgtcagcc agtttagtct gaccatctca tctgtaacat cattggcaac gctacctttg
     7441 ccatgtttca gaaacaactc tggcgcatcg ggcttcccat acaatcggta gattgtcgca
     7501 cctgattgcc cgacattatc gcgagcccat ttatacccat ataaatcagc atccatgttg
     7561 gaatttaatc gcggccttga gcaagacgtt tcccgttgaa tatggctcat aacagaactt
     7621 attatttcct tcctcttttc tacagtattt aaagataccc caagaagcta attataacaa
     7681 gacgaactcc aattcactgt tccttgcatt ctaaaacctt aaataccaga aaacagcttt
     7741 ttcaaagttg ttttcaaagt tggcgtataa catagtatcg acggagccga ttttgaaacc
     7801 gcggtgatca caggcagcaa cgctctgtca tcgttacaat caacatgcta ccctccgcga
     7861 gatcatccgt gtttcaaacc cggcagctta gttgccgttc ttccgaatag catcggtaac
     7921 atgagcaaag tctgccgcct tacaacggct ctcccgctga cgccgtcccg gactgatggg
     7981 ctgcctgtat cgagtggtga ttttgtgccg agctgccggt cggggagctg ttggctggct
     8041 gg
//
