LOCUS       sequence-405991-        7872 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TU for the expression of phleomycin resistance..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7872)
  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 7872)
  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: FB430
FEATURES             Location/Qualifiers
     source          1..7872
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             769..1143
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     misc_feature    1516..1540
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             2841..3470
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             3899..4972
                     /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      5038..5232
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(5468..5490)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    5576..5716
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     5631..5650
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5731..5748)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5902..6490)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5982..6001)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6578..7393)
                     /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(6578..7393)
                     /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     7301..7320
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     misc_feature    7826..7850
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
ORIGIN
        1 ggagggtcga ctacatgtat ctgcatgttg catcgggaaa tcccaccaca gggacagcca
       61 agcggccccg cgacttggca gtgggcaaac tacgcccgat tctggtgcca agaaccgaga
      121 agaatgagac agacccacgt tgcactctaa ccggatgcta tcgacttacg gtggctgaag
      181 attcaacacg ctgcaacgag agccaaggtg gtccggacat tttctacgtg ccggtttacc
      241 ttggaacatc gccgtcgttg agtgcacgtt gcctactctc tcgtggcttg gctgggccca
      301 cgagcccgat tgactcgacg gtgttacttg ggtatctatg gccccgtttt ctggcacggt
      361 aatgataagt acttactagt cttcgagcga gcgggggagt gttgctctgc ccgagcatca
      421 acgattggcc tgatcgcacc gtctgcaaat gccacggtgc ggaccgactg aaatctcaga
      481 ccaccaaaga ccctccgact tcgagatacg gttactaatt ttacactggc tccagcggcc
      541 ccatccagta agcatctggg ctgcaagcgt ataatgtctc caggttgtct cagcataaac
      601 accccgcccc cgctcaggca cacaggaaga gagctcaggt cgtttccatt gcgtccatac
      661 tcttcactca ttgtcatctg caggagaact tcccctgtcc ctttgccaag ccctctcttc
      721 gtcgttgtcc acgccttcaa gttttcacca ttatttttct agacaccaat ggccaagttg
      781 accagtgccg ttccggtgct caccgcgcgc gacgtcgccg gagcggtcga gttctggacc
      841 gaccggctcg ggttctcccg ggacttcgtg gaggacgact tcgccggtgt ggtccgggac
      901 gacgtgaccc tgttcatcag cgcggtccag gaccaggtgg tgccggacaa caccctggcc
      961 tgggtgtggg tgcgcggcct ggacgagctg tacgccgagt ggtcggaggt cgtgtccacg
     1021 aacttccggg acgcctccgg gccggccatg accgagatcg gcgagcagcc gtgggggcgg
     1081 gagttcgccc tgcgcgaccc ggccggcaac tgcgtgcact tcgtggccga ggagcaggac
     1141 tgagcttcta ataagtgtca gatagcaatt tgcacaagaa atcaatacca gcaactgtaa
     1201 ataagcgctg aagtgaccat gccatgctac gaaagagcag aaaaaaacct gccgtagaac
     1261 cgaagagata tgacacgctt ccatctctca aaggaagaat cccttcaggg ttgcgtttcc
     1321 agtctagaca cgtataacgg cacaagtgtc tctcaccaaa tgggttatat ctcaaatgtg
     1381 atctaaggat ggaaagccca gaatattggc tgggttgatg gctgcttcga gtgcagtctc
     1441 atgctgccac aggtgactct ggatggcccc ataccactca acccatggta cgagcggtac
     1501 gcttgagacg aagcttgaca ggatatattg gcgggtaaac ctaagagaaa agagcgttta
     1561 ttagaataat cggatattta aaagggcgtg aaaaggttta tccgttcgtc catttgtatg
     1621 tgcatgccaa ccacagggtt cccctcggga tcaaagtact ttgatccaac ccctccgctg
     1681 ctatagtgca gtcggcttct gacgttcagt gcagccgtca tctgaaaacg acatgtcgca
     1741 caagtcctaa gttacgcgac aggctgccgc cctgcccttt tcctggcgtt ttcttgtcgc
     1801 gtgttttagt cgcataaagt agaatacttg cgactagaac cggagacatt acgccatgaa
     1861 caagagcgcc gccgctggcc tgctgggcta tgcccgcgtc agcaccgacg accaggactt
     1921 gaccaaccaa cgggccgaac tgcacgcggc cggctgcacc aagctgtttt ccgagaagat
     1981 caccggcacc aggcgcgacc gcccggagct ggccaggatg cttgaccacc tacgccctgg
     2041 cgacgttgtg acagtgacca ggctagaccg cctggcccgc agcacccgcg acctactgga
     2101 cattgccgag cgcatccagg aggccggcgc gggcctgcgt agcctggcag agccgtgggc
     2161 cgacaccacc acgccggccg gccgcatggt gttgaccgtg ttcgccggca ttgccgagtt
     2221 cgagcgttcc ctaatcatcg accgcacccg gagcgggcgc gaggccgcca aggcccgagg
     2281 cgtgaagttt ggcccccgcc ctaccctcac cccggcacag atcgcgcacg cccgcgagct
     2341 gatcgaccag gaaggccgca ccgtgaaaga ggcggctgca ctgcttggcg tgcatcgctc
     2401 gaccctgtac cgcgcacttg agcgcagcga ggaagtgacg cccaccgagg ccaggcggcg
     2461 cggtgccttc cgtgaggacg cattgaccga ggccgacgcc ctggcggccg ccgagaatga
     2521 acgccaagag gaacaagcat gaaaccgcac caggacggcc aggacgaacc gtttttcatt
     2581 accgaagaga tcgaggcgga gatgatcgcg gccgggtacg tgttcgagcc gcccgcgcac
     2641 ctctcaaccg tgcggctgca tgaaatcctg gccggtttgt ctgatgccaa gctggcggcc
     2701 tggccggcca gcttggccgc tgaagaaacc gagcgccgcc gtctaaaaag gtgatgtgta
     2761 tttgagtaaa acagcttgcg tcatgcggtc gctgcgtata tgatgcgatg agtaaataaa
     2821 caaatacgca aggggaacgc atgaaggtta tcgctgtact taaccagaaa ggcgggtcag
     2881 gcaagacgac catcgcaacc catctagccc gcgccctgca actcgccggg gccgatgttc
     2941 tgttagtcga ttccgatccc cagggcagtg cccgcgattg ggcggccgtg cgggaagatc
     3001 aaccgctaac cgttgtcggc atcgaccgcc cgacgattga ccgcgacgtg aaggccatcg
     3061 gccggcgcga cttcgtagtg atcgacggag cgccccaggc ggcggacttg gctgtgtccg
     3121 cgatcaaggc agccgacttc gtgctgattc cggtgcagcc aagcccttac gacatatggg
     3181 ccaccgccga cctggtggag ctggttaagc agcgcattga ggtcacggat ggaaggctac
     3241 aagcggcctt tgtcgtgtcg cgggcgatca aaggcacgcg catcggcggt gaggttgccg
     3301 aggcgctggc cgggtacgag ctgcccattc ttgagtcccg tatcacgcag cgcgtgagct
     3361 acccaggcac tgccgccgcc ggcacaaccg ttcttgaatc agaacccgag ggcgacgctg
     3421 cccgcgaggt ccaggcgctg gccgctgaaa ttaaatcaaa actcatttga gttaatgagg
     3481 taaagagaaa atgagcaaaa gcacaaacac gctaagtgcc ggccgtccga gcgcacgcag
     3541 cagcaaggct gcaacgttgg ccagcctggc agacacgcca gccatgaagc gggtcaactt
     3601 tcagttgccg gcggaggatc acaccaagct gaagatgtac gcggtacgcc aaggcaagac
     3661 cattaccgag ctgctatctg aatacatcgc gcagctacca gagtaaatga gcaaatgaat
     3721 aaatgagtag atgaatttta gcggctaaag gaggcggcat ggaaaatcaa gaacaaccag
     3781 gcaccgacgc cgtggaatgc cccatgtgtg gaggaacggg cggttggcca ggcgtaagcg
     3841 gctgggttgc ctgccggccc tgcaatggca ctggaacccc caagcccgag gaatcggcgt
     3901 gagcggtcgc aaaccatccg gcccggtaca aatcggcgcg gcgctgggtg atgacctggt
     3961 ggagaagttg aaggccgcgc aggccgccca gcggcaacgc atcgaggcag aagcacgccc
     4021 cggtgaatcg tggcaagcgg ccgctgatcg aatccgcaaa gaatcccggc aaccgccggc
     4081 agccggtgcg ccgtcgatta ggaagccgcc caagggcgac gagcaaccag attttttcgt
     4141 tccgatgctc tatgacgtgg gcacccgcga tagtcgcagc atcatggacg tggccgtttt
     4201 ccgtctgtcg aagcgtgacc gacgagctgg cgaggtgatc cgctacgagc ttccagacgg
     4261 gcacgtagag gtttccgcag ggccggccgg catggccagt gtgtgggatt acgacctggt
     4321 actgatggcg gtttcccatc taaccgaatc catgaaccga taccgggaag ggaagggaga
     4381 caagcccggc cgcgtgttcc gtccacacgt tgcggacgta ctcaagttct gccggcgagc
     4441 cgatggcgga aagcagaaag acgacctggt agaaacctgc attcggttaa acaccacgca
     4501 cgttgccatg cagcgtacga agaaggccaa gaacggccgc ctggtgacgg tatccgaggg
     4561 tgaagccttg attagccgct acaagatcgt aaagagcgaa accgggcggc cggagtacat
     4621 cgagatcgag ctagctgatt ggatgtaccg cgagatcaca gaaggcaaga acccggacgt
     4681 gctgacggtt caccccgatt actttttgat cgatcccggc atcggccgtt ttctctaccg
     4741 cctggcacgc cgcgccgcag gcaaggcaga agccagatgg ttgttcaaga cgatctacga
     4801 acgcagtggc agcgccggag agttcaagaa gttctgtttc accgtgcgca agctgatcgg
     4861 gtcaaatgac ctgccggagt acgatttgaa ggaggaggcg gggcaggctg gcccgatcct
     4921 agtcatgcgc taccgcaacc tgatcgaggg cgaagcatcc gccggttcct aatgtacgga
     4981 gcagatgcta gggcaaattg ccctagcagg ggaaaaaggt cgaaaaggac tctttcctgt
     5041 ggatagcacg tacattggga acccaaagcc gtacattggg aaccggaacc cgtacattgg
     5101 gaacccaaag ccgtacattg ggaaccggtc acacatgtaa gtgactgata taaaagagaa
     5161 aaaaggcgat ttttccgcct aaaactcttt aaaacttatt aaaactctta aaacccgcct
     5221 ggcctgtgca taactgtctg gccagcgcac agccgaagag ctgcaaaaag cgcctaccct
     5281 tcggtcgctg cgctccctac gccccgccgc ttcgcgtcgg cctatcgcgg ccgctggccg
     5341 ctcaaaaatg gctggcctac ggccaggcaa tctaccaggg cgcggacaag ccgcgccgtc
     5401 gccactcgac cgccggcgcc cacatcaagg caccctgcct cgcgcgtttc ggtgatgacg
     5461 gtgaaaacct ctgacacatg cagctcccgg tgacggtcac agcttgtctg taagcggatg
     5521 ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag
     5581 ccatgaccca gtcacgtagc gatagcggag tgtatactgg cttaactatg cggcatcaga
     5641 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag
     5701 aaaataccgc atcaggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     5761 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     5821 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     5881 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     5941 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     6001 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     6061 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     6121 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     6181 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     6241 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     6301 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
     6361 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     6421 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     6481 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     6541 ctcacgttaa gggattttgg tcatgcattc taggtgatta gaaaaactca tcgagcatca
     6601 aatgaaactg caatttattc atatcaggat tatcaatacc atatttttga aaaagccgtt
     6661 tctgtaatga aggagaaaac tcaccgaggc agttccatag gatggcaaga tcctggtatc
     6721 ggtctgcgat tccgactcgt ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa
     6781 taaggttatc aagtgagaaa tcaccatgag tgacgactga atccggtgag aatggcaaaa
     6841 gtttatgcat ttctttccag acttgttcaa caggccagcc attacgctcg tcatcaaaat
     6901 cactcgcatc aaccaaaccg ttattcattc gtgattgcgc ctgagcgagt cgaaatacgc
     6961 gatcgctgtt aaaaggacaa ttacaaacag gaatcgaatg caaccggcgc aggaacactg
     7021 ccagcgcatc aacaatattt tcacctgaat caggatattc ttctaatacc tggaatgctg
     7081 ttttccctgg gatcgcagtg gtgagtaacc atgcatcatc aggagtacgg ataaaatgct
     7141 tgatggtcgg aagaggcata aattccgtca gccagtttag tctgaccatc tcatctgtaa
     7201 catcattggc aacgctacct ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc
     7261 catacaatcg gtagattgtc gcacctgatt gcccgacatt atcgcgagcc catttatacc
     7321 catataaatc agcatccatg ttggaattta atcgcggcct tgagcaagac gtttcccgtt
     7381 gaatatggct cataacagaa cttattattt ccttcctctt ttctacagta tttaaagata
     7441 ccccaagaag ctaattataa caagacgaac tccaattcac tgttccttgc attctaaaac
     7501 cttaaatacc agaaaacagc tttttcaaag ttgttttcaa agttggcgta taacatagta
     7561 tcgacggagc cgattttgaa accgcggtga tcacaggcag caacgctctg tcatcgttac
     7621 aatcaacatg ctaccctccg cgagatcatc cgtgtttcaa acccggcagc ttagttgccg
     7681 ttcttccgaa tagcatcggt aacatgagca aagtctgccg ccttacaacg gctctcccgc
     7741 tgacgccgtc ccggactgat gggctgcctg tatcgagtgg tgattttgtg ccgagctgcc
     7801 ggtcggggag ctgttggctg gctggtggca ggatatattg tggtgtaaac ataacaagct
     7861 tcgtctcagt ca
//
