LOCUS       sequence-236967-        7812 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Transcriptional Unit (TU) for geneticin (G418) resistance in fungi 
            obtained by combining FB001+FB005+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 7812)
  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 7812)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: FB009
FEATURES             Location/Qualifiers
     source          1..7812
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    83..107
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             496..1290
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(550..569)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1160..1179
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     misc_feature    1585..1609
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             2910..3539
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             3968..5041
                     /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      5107..5301
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(5537..5559)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    5645..5785
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     5700..5719
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5800..5817)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5971..6559)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6051..6070)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6647..7462)
                     /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(6647..7462)
                     /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     7370..7389
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 cgccgtcccg gactgatggg ctgcctgtat cgagtggtga ttttgtgccg agctgccggt
       61 cggggagctg ttggctggct ggtggcagga tatattgtgg tgtaaacata acaagcttcg
      121 tctcagtcag gagaactgat attgaaggag cattttttgg gcttggctgg agctagtgga
      181 ggtcaacaat gaatgcctat tttggtttag tcgtccaggc ggtgagcaca aaatttgtgt
      241 cgtttgacaa gatggttcat ttaggcaact ggtcagatca gccccacttg tagcagtagc
      301 ggcggcgctc gaagtgtgac tcttattagc agacaggaac gaggacatta ttatcatctg
      361 ctgcttggtg cacgataact tggtgcgttt gtcaagcaag gtaagtggac gacccggtca
      421 taccttctta agttcgccct tcctcccttt atttcagatt caatctgact tacctattct
      481 acccaagcat ccaaaatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
      541 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
      601 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
      661 gccctgaatg aactccaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
      721 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
      781 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
      841 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
      901 caagcgaaac accgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
      961 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     1021 gcgcggatgc ccgacggcga ggatctcgtc gtgacccacg gcgacgcctg cttgccgaat
     1081 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     1141 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     1201 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatagcc
     1261 ttctatcgcc ttcttgacga gttcttctga gcttcatcat tccaccatca ttccactcaa
     1321 cattcaggct cctctgcgca cgtaaagtgc caaaggcaat accctgctcg gtggaatgcc
     1381 gccgggcttg tcgattttac gcacatatgc gcattcttga cttgaagcgg aggagttctt
     1441 cgttgcgggt tacagtgttt taataaaaga atggtcaaat caaactgcta gatatacctg
     1501 tcagacactc tagttgttga cccctatact cttaatacat cagacagtac atgcatgttg
     1561 catgatgacg cttgagacga agcttgacag gatatattgg cgggtaaacc taagagaaaa
     1621 gagcgtttat tagaataatc ggatatttaa aagggcgtga aaaggtttat ccgttcgtcc
     1681 atttgtatgt gcatgccaac cacagggttc ccctcgggat caaagtactt tgatccaacc
     1741 cctccgctgc tatagtgcag tcggcttctg acgttcagtg cagccgtcat ctgaaaacga
     1801 catgtcgcac aagtcctaag ttacgcgaca ggctgccgcc ctgccctttt cctggcgttt
     1861 tcttgtcgcg tgttttagtc gcataaagta gaatacttgc gactagaacc ggagacatta
     1921 cgccatgaac aagagcgccg ccgctggcct gctgggctat gcccgcgtca gcaccgacga
     1981 ccaggacttg accaaccaac gggccgaact gcacgcggcc ggctgcacca agctgttttc
     2041 cgagaagatc accggcacca ggcgcgaccg cccggagctg gccaggatgc ttgaccacct
     2101 acgccctggc gacgttgtga cagtgaccag gctagaccgc ctggcccgca gcacccgcga
     2161 cctactggac attgccgagc gcatccagga ggccggcgcg ggcctgcgta gcctggcaga
     2221 gccgtgggcc gacaccacca cgccggccgg ccgcatggtg ttgaccgtgt tcgccggcat
     2281 tgccgagttc gagcgttccc taatcatcga ccgcacccgg agcgggcgcg aggccgccaa
     2341 ggcccgaggc gtgaagtttg gcccccgccc taccctcacc ccggcacaga tcgcgcacgc
     2401 ccgcgagctg atcgaccagg aaggccgcac cgtgaaagag gcggctgcac tgcttggcgt
     2461 gcatcgctcg accctgtacc gcgcacttga gcgcagcgag gaagtgacgc ccaccgaggc
     2521 caggcggcgc ggtgccttcc gtgaggacgc attgaccgag gccgacgccc tggcggccgc
     2581 cgagaatgaa cgccaagagg aacaagcatg aaaccgcacc aggacggcca ggacgaaccg
     2641 tttttcatta ccgaagagat cgaggcggag atgatcgcgg ccgggtacgt gttcgagccg
     2701 cccgcgcacc tctcaaccgt gcggctgcat gaaatcctgg ccggtttgtc tgatgccaag
     2761 ctggcggcct ggccggccag cttggccgct gaagaaaccg agcgccgccg tctaaaaagg
     2821 tgatgtgtat ttgagtaaaa cagcttgcgt catgcggtcg ctgcgtatat gatgcgatga
     2881 gtaaataaac aaatacgcaa ggggaacgca tgaaggttat cgctgtactt aaccagaaag
     2941 gcgggtcagg caagacgacc atcgcaaccc atctagcccg cgccctgcaa ctcgccgggg
     3001 ccgatgttct gttagtcgat tccgatcccc agggcagtgc ccgcgattgg gcggccgtgc
     3061 gggaagatca accgctaacc gttgtcggca tcgaccgccc gacgattgac cgcgacgtga
     3121 aggccatcgg ccggcgcgac ttcgtagtga tcgacggagc gccccaggcg gcggacttgg
     3181 ctgtgtccgc gatcaaggca gccgacttcg tgctgattcc ggtgcagcca agcccttacg
     3241 acatatgggc caccgccgac ctggtggagc tggttaagca gcgcattgag gtcacggatg
     3301 gaaggctaca agcggccttt gtcgtgtcgc gggcgatcaa aggcacgcgc atcggcggtg
     3361 aggttgccga ggcgctggcc gggtacgagc tgcccattct tgagtcccgt atcacgcagc
     3421 gcgtgagcta cccaggcact gccgccgccg gcacaaccgt tcttgaatca gaacccgagg
     3481 gcgacgctgc ccgcgaggtc caggcgctgg ccgctgaaat taaatcaaaa ctcatttgag
     3541 ttaatgaggt aaagagaaaa tgagcaaaag cacaaacacg ctaagtgccg gccgtccgag
     3601 cgcacgcagc agcaaggctg caacgttggc cagcctggca gacacgccag ccatgaagcg
     3661 ggtcaacttt cagttgccgg cggaggatca caccaagctg aagatgtacg cggtacgcca
     3721 aggcaagacc attaccgagc tgctatctga atacatcgcg cagctaccag agtaaatgag
     3781 caaatgaata aatgagtaga tgaattttag cggctaaagg aggcggcatg gaaaatcaag
     3841 aacaaccagg caccgacgcc gtggaatgcc ccatgtgtgg aggaacgggc ggttggccag
     3901 gcgtaagcgg ctgggttgcc tgccggccct gcaatggcac tggaaccccc aagcccgagg
     3961 aatcggcgtg agcggtcgca aaccatccgg cccggtacaa atcggcgcgg cgctgggtga
     4021 tgacctggtg gagaagttga aggccgcgca ggccgcccag cggcaacgca tcgaggcaga
     4081 agcacgcccc ggtgaatcgt ggcaagcggc cgctgatcga atccgcaaag aatcccggca
     4141 accgccggca gccggtgcgc cgtcgattag gaagccgccc aagggcgacg agcaaccaga
     4201 ttttttcgtt ccgatgctct atgacgtggg cacccgcgat agtcgcagca tcatggacgt
     4261 ggccgttttc cgtctgtcga agcgtgaccg acgagctggc gaggtgatcc gctacgagct
     4321 tccagacggg cacgtagagg tttccgcagg gccggccggc atggccagtg tgtgggatta
     4381 cgacctggta ctgatggcgg tttcccatct aaccgaatcc atgaaccgat accgggaagg
     4441 gaagggagac aagcccggcc gcgtgttccg tccacacgtt gcggacgtac tcaagttctg
     4501 ccggcgagcc gatggcggaa agcagaaaga cgacctggta gaaacctgca ttcggttaaa
     4561 caccacgcac gttgccatgc agcgtacgaa gaaggccaag aacggccgcc tggtgacggt
     4621 atccgagggt gaagccttga ttagccgcta caagatcgta aagagcgaaa ccgggcggcc
     4681 ggagtacatc gagatcgagc tagctgattg gatgtaccgc gagatcacag aaggcaagaa
     4741 cccggacgtg ctgacggttc accccgatta ctttttgatc gatcccggca tcggccgttt
     4801 tctctaccgc ctggcacgcc gcgccgcagg caaggcagaa gccagatggt tgttcaagac
     4861 gatctacgaa cgcagtggca gcgccggaga gttcaagaag ttctgtttca ccgtgcgcaa
     4921 gctgatcggg tcaaatgacc tgccggagta cgatttgaag gaggaggcgg ggcaggctgg
     4981 cccgatccta gtcatgcgct accgcaacct gatcgagggc gaagcatccg ccggttccta
     5041 atgtacggag cagatgctag ggcaaattgc cctagcaggg gaaaaaggtc gaaaaggact
     5101 ctttcctgtg gatagcacgt acattgggaa cccaaagccg tacattggga accggaaccc
     5161 gtacattggg aacccaaagc cgtacattgg gaaccggtca cacatgtaag tgactgatat
     5221 aaaagagaaa aaaggcgatt tttccgccta aaactcttta aaacttatta aaactcttaa
     5281 aacccgcctg gcctgtgcat aactgtctgg ccagcgcaca gccgaagagc tgcaaaaagc
     5341 gcctaccctt cggtcgctgc gctccctacg ccccgccgct tcgcgtcggc ctatcgcggc
     5401 cgctggccgc tcaaaaatgg ctggcctacg gccaggcaat ctaccagggc gcggacaagc
     5461 cgcgccgtcg ccactcgacc gccggcgccc acatcaaggc accctgcctc gcgcgtttcg
     5521 gtgatgacgg tgaaaacctc tgacacatgc agctcccggt gacggtcaca gcttgtctgt
     5581 aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc
     5641 ggggcgcagc catgacccag tcacgtagcg atagcggagt gtatactggc ttaactatgc
     5701 ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg
     5761 cgtaaggaga aaataccgca tcaggcgctc ttccgcttcc tcgctcactg actcgctgcg
     5821 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     5881 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     5941 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     6001 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     6061 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     6121 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     6181 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     6241 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     6301 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     6361 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     6421 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     6481 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     6541 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     6601 gaacgaaaac tcacgttaag ggattttggt catgcattct aggtgattag aaaaactcat
     6661 cgagcatcaa atgaaactgc aatttattca tatcaggatt atcaatacca tatttttgaa
     6721 aaagccgttt ctgtaatgaa ggagaaaact caccgaggca gttccatagg atggcaagat
     6781 cctggtatcg gtctgcgatt ccgactcgtc caacatcaat acaacctatt aatttcccct
     6841 cgtcaaaaat aaggttatca agtgagaaat caccatgagt gacgactgaa tccggtgaga
     6901 atggcaaaag tttatgcatt tctttccaga cttgttcaac aggccagcca ttacgctcgt
     6961 catcaaaatc actcgcatca accaaaccgt tattcattcg tgattgcgcc tgagcgagtc
     7021 gaaatacgcg atcgctgtta aaaggacaat tacaaacagg aatcgaatgc aaccggcgca
     7081 ggaacactgc cagcgcatca acaatatttt cacctgaatc aggatattct tctaatacct
     7141 ggaatgctgt tttccctggg atcgcagtgg tgagtaacca tgcatcatca ggagtacgga
     7201 taaaatgctt gatggtcgga agaggcataa attccgtcag ccagtttagt ctgaccatct
     7261 catctgtaac atcattggca acgctacctt tgccatgttt cagaaacaac tctggcgcat
     7321 cgggcttccc atacaatcgg tagattgtcg cacctgattg cccgacatta tcgcgagccc
     7381 atttataccc atataaatca gcatccatgt tggaatttaa tcgcggcctt gagcaagacg
     7441 tttcccgttg aatatggctc ataacagaac ttattatttc cttcctcttt tctacagtat
     7501 ttaaagatac cccaagaagc taattataac aagacgaact ccaattcact gttccttgca
     7561 ttctaaaacc ttaaatacca gaaaacagct ttttcaaagt tgttttcaaa gttggcgtat
     7621 aacatagtat cgacggagcc gattttgaaa ccgcggtgat cacaggcagc aacgctctgt
     7681 catcgttaca atcaacatgc taccctccgc gagatcatcc gtgtttcaaa cccggcagct
     7741 tagttgccgt tcttccgaat agcatcggta acatgagcaa agtctgccgc cttacaacgg
     7801 ctctcccgct ga
//
