LOCUS       sequence-310999-        7019 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  pEGB3a1 AtS/MAR10.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 7019)
  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 7019)
  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: AtS/MAR10 insulator sequence (GB3458)
FEATURES             Location/Qualifiers
     source          1..7019
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     misc_feature    669..693
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             1994..2623
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIGTHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             3057..4124
                     /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      4190..4384
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(4620..4642)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4728..4868
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4783..4802
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4883..4900)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5054..5642)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5134..5153)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5730..6545)
                     /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(5730..6545)
                     /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     6453..6472
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     misc_feature    6978..7002
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
ORIGIN
        1 ggagtggcta ttgttgttat catcaaaatc tatattgata ctattatact cgatatgtta
       61 tagactttat taaaaaaaac tttgtttgtt tgcagaaagt ggatttttaa tcaaatttaa
      121 ttgtgatgat tacaatgtca attggttatg caagatatta attaatatgt ttaagatttt
      181 tcacttttaa taagtaaaat aaaatacgtc aaaaaaattt cctaaagata atgccaaaaa
      241 ggttaaatta tagaaattaa actaaaatat attaaaagtt gacttaaaaa tgcaatgttg
      301 accaccgacc taggggaaaa caaatttacc gtatctggga tagaatctct tctgtatcat
      361 ctcagataaa gattactaca taaacacact ttacagtttt acgacgccca aaattcaatt
      421 cccggagatt aaaaacccta aaacaaacac tggcgattcg attttccctc ccaattcaaa
      481 tcatctctta atcttcttct tctcactatc ctctcaaatt cgccgccgat cttccatcaa
      541 ttttgatttc tcgcaccgaa atttcgactt ttcatctcct atcgacgaaa ccctaatttt
      601 tcacactcca gaaatcgaac tttgacgacg atgttaatgg ctaaaccccg ctgtcatgag
      661 acgaattctg acaggatata ttggcgggta aacctaagag aaaagagcgt ttattagaat
      721 aatcggatat ttaaaagggc gtgaaaaggt ttatccgttc gtccatttgt atgtgcatgc
      781 caaccacagg gttcccctcg ggatcaaagt actttgatcc aacccctccg ctgctatagt
      841 gcagtcggct tctgacgttc agtgcagccg tcatctgaaa acgacatgtc gcacaagtcc
      901 taagttacgc gacaggctgc cgccctgccc ttttcctggc gttttcttgt cgcgtgtttt
      961 agtcgcataa agtagaatac ttgcgactag aaccggagac attacgccat gaacaagagc
     1021 gccgccgctg gcctgctggg ctatgcccgc gtcagcaccg acgaccagga cttgaccaac
     1081 caacgggccg aactgcacgc ggccggctgc accaagctgt tttccgagaa gatcaccggc
     1141 accaggcgcg accgcccgga gctggccagg atgcttgacc acctacgccc tggcgacgtt
     1201 gtgacagtga ccaggctaga ccgcctggcc cgcagcaccc gcgacctact ggacattgcc
     1261 gagcgcatcc aggaggccgg cgcgggcctg cgtagcctgg cagagccgtg ggccgacacc
     1321 accacgccgg ccggccgcat ggtgttgacc gtgttcgccg gcattgccga gttcgagcgt
     1381 tccctaatca tcgaccgcac ccggagcggg cgcgaggccg ccaaggcccg aggcgtgaag
     1441 tttggccccc gccctaccct caccccggca cagatcgcgc acgcccgcga gctgatcgac
     1501 caggaaggcc gcaccgtgaa agaggcggct gcactgcttg gcgtgcatcg ctcgaccctg
     1561 taccgcgcac ttgagcgcag cgaggaagtg acgcccaccg aggccaggcg gcgcggtgcc
     1621 ttccgtgagg acgcattgac cgaggccgac gccctggcgg ccgccgagaa tgaacgccaa
     1681 gaggaacaag catgaaaccg caccaggacg gccaggacga accgtttttc attaccgaag
     1741 agatcgaggc ggagatgatc gcggccgggt acgtgttcga gccgcccgcg cacctctcaa
     1801 ccgtgcggct gcatgaaatc ctggccggtt tgtctgatgc caagctggcg gcctggccgg
     1861 ccagcttggc cgctgaagaa accgagcgcc gccgtctaaa aaggtgatgt gtatttgagt
     1921 aaaacagctt gcgtcatgcg gtcgctgcgt atatgatccg atgagtaaat aaacaaatac
     1981 gcaaggggaa cgcatgaagg ttatcgctgt acttaaccag aaaggcgggt caggcaagac
     2041 gaccatcgga acccatctag cccgcgccct gcaactcgcc ggggccgatg ttctgttagt
     2101 cgattccgat ccccagggca gtgcccgcga ttgggcggcc gtgcgggaag atcaaccgct
     2161 aaccgttgtc ggcatcgacc gcccgacgat tgaccgcgac gtgaaggcca tcggccggcg
     2221 cgacttcgta gtgatcgacg gagcgcccca ggcggcggac ttggctgtgt ccgcgatcaa
     2281 ggcagccgac ttcgtgctga ttccggtgca gccaagccct tacgacatat gggccaccgc
     2341 cgacctggtg gagctggtta agcagcgcat tgaggtcacg gatggaaggc tacaagcggc
     2401 ctttgtcgtg tcgcgggcga tcaaaggcac gcgcatcggc ggtgaggttg ccgaggcgct
     2461 ggccgggtac gagctgccca ttcttgagtc ccgtatcacg cagcgcgtga gctacccagg
     2521 cactgccgcc gccggcacaa ccgttcttga atcagaaccc gagggcgacg ctgcccgcga
     2581 ggtccaggcg ctggccgctg aaattaaatc aaaactcatt tgagttaatg aggtaaagag
     2641 aaaatgagca aaagcacaaa cacgctaagt gccggccgtc cgagcgcacg cagcagcaag
     2701 gctgcaacgt tggccagcct ggcagacacg ccagccatga agcgggtcaa ctttcagttg
     2761 ccggcggagg atcacaccaa gctgaagatg tacgcggtac gccaaggcaa gaccattacc
     2821 gagctgctat ctgaatagat cgcgcagcta ccagagtaaa tgagcaaatg aataaatgag
     2881 tagatgaatt ttagcggcta aaggaggcgg catggaaaat caagaacaac caggcaccga
     2941 cgccgtggaa tgccccatgt gtggaggaac gggcggttgg ccaggcgtaa gcggctgggt
     3001 tgtctgccgg ccctgcaatg gcactggaac ccccaagccc gaggaatcgg cgtgacggtc
     3061 gcaaaccatc cggcccggta caaatcggcg cggcgctggg tgatgacctg gtggagaagt
     3121 tgaaggccgc gcaggccgcc cagcggcaac gcatcgaggc agaagcacgc cccggtgaat
     3181 cgtggcaagc ggccgctgat cgaatccgca aagaatcccg gcaaccgccg gcagccggtg
     3241 cgccgtcgat taggaagccg cccaagggcg acgagcaacc agattttttc gttccgatgc
     3301 tctatgacgt gggcacccgc gatagtcgca gcatcatgga cgtggccgtt ttccgtctgt
     3361 cgaagcgtga ccgacgagct ggcgaggtga tccgctacga gcttccagac gggcacgtag
     3421 aggtttccgc agggccggcc ggcatggcca gtgtgtggga ttacgacctg gtactgatgg
     3481 cggtttccca tctaaccgaa tccatgaacc gataccggga agggaaggga gacaagcccg
     3541 gccgcgtgtt ccgtccacac gttgcggacg tactcaagtt ctgccggcga gccgatggcg
     3601 gaaagcagaa agacgacctg gtagaaacct gcattcggtt aaacaccacg cacgttgcca
     3661 tgcagcgtac gaagaaggcc aagaacggcc gcctggtgac ggtatccgag ggtgaagcct
     3721 tgattagccg ctacaagatc gtaaagagcg aaaccgggcg gccggagtac atcgagatcg
     3781 agctagctga ttggatgtac cgcgagatca cagaaggcaa gaacccggac gtgctgacgg
     3841 ttcaccccga ttactttttg atcgatcccg gcatcggccg ttttctctac cgcctggcac
     3901 gccgcgccgc aggcaaggca gaagccagat ggttgttcaa gacgatctac gaacgcagtg
     3961 gcagcgccgg agagttcaag aagttctgtt tcaccgtgcg caagctgatc gggtcaaatg
     4021 acctgccgga gtacgatttg aaggaggagg cggggcaggc tggcccgatc ctagtcatgc
     4081 gctaccgcaa cctgatcgag ggcgaagcat ccgccggttc ctaatgtacg gagcagatgc
     4141 tagggcaaat tgccctagca ggggaaaaag gtcgaaaagg actctttcct gtggatagca
     4201 cgtacattgg gaacccaaag ccgtacattg ggaaccggaa cccgtacatt gggaacccaa
     4261 agccgtacat tgggaaccgg tcacacatgt aagtgactga tataaaagag aaaaaaggcg
     4321 atttttccgc ctaaaactct ttaaaactta ttaaaactct taaaacccgc ctggcctgtg
     4381 cataactgtc tggccagcgc acagccgaag agctgcaaaa agcgcctacc cttcggtcgc
     4441 tgcgctccct acgccccgcc gcttcgcgtc ggcctatcgc ggccgctggc cgctcaaaaa
     4501 tggctggcct acggccaggc aatctaccag ggcgcggaca agccgcgccg tcgccactcg
     4561 accgccggcg cccacatcaa ggcaccctgc ctcgcgcgtt tcggtgatga cggtgaaaac
     4621 ctctgacaca tgcagctccc ggtgacggtc acagcttgtc tgtaagcgga tgccgggagc
     4681 agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc agccatgacc
     4741 cagtcacgta gcgatagcgg agtgtatact ggcttaacta tgcggcatca gagcagattg
     4801 tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc
     4861 gcatcaggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     4921 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     4981 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     5041 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     5101 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     5161 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     5221 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     5281 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     5341 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     5401 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     5461 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
     5521 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     5581 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     5641 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     5701 aagggatttt ggtcatgcat tctaggtgat tagaaaaact catcgagcat caaatgaaac
     5761 tgcaatttat tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat
     5821 gaaggagaaa actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg
     5881 attccgactc gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta
     5941 tcaagtgaga aatcaccatg agtgacgact gaatccggtg agaatggcaa aagtttatgc
     6001 atttctttcc agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca
     6061 tcaaccaaac cgttattcat tcgtgattgc gcctgagcga gtcgaaatac gcgatcgctg
     6121 ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca
     6181 tcaacaatat tttcacctga atcaggatat tcttctaata cctggaatgc tgttttccct
     6241 gggatcgcag tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc
     6301 ggaagaggca taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg
     6361 gcaacgctac ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaat
     6421 cggtagattg tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa
     6481 tcagcatcca tgttggaatt taatcgcggc cttgagcaag acgtttcccg ttgaatatgg
     6541 ctcataacag aacttattat ttccttcctc ttttctacag tatttaaaga taccccaaga
     6601 agctaattat aacaagacga actccaattc actgttcctt gcattctaaa accttaaata
     6661 ccagaaaaca gctttttcaa agttgttttc aaagttggcg tataacatag tatcgacgga
     6721 gccgattttg aaaccgcggt gatcacaggc agcaacgctc tgtcatcgtt acaatcaaca
     6781 tgctaccctc cgcgagatca tccgtgtttc aaacccggca gcttagttgc cgttcttccg
     6841 aatagcatcg gtaacatgag caaagtctgc cgccttacaa cggctctccc gctgacgccg
     6901 tcccggactg atgggctgcc tgtatcgagt ggtgattttg tgccgagctg ccggtcgggg
     6961 agctgttggc tggctggtgg caggatatat tgtggtgtaa acataacgaa ttcgtctca
//
