LOCUS       sequence-321959-        7020 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pEGB3a1 AtS/MAR10.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7020)
  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 7020)
  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..7020
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             3052..4125
                     /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      4191..4385
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(4621..4643)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4729..4869
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4784..4803
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4884..4901)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5055..5643)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5135..5154)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5731..6546)
                     /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(5731..6546)
                     /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     6454..6473
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     misc_feature    6979..7003
                     /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 atatgatgcg atgagtaaat aaacaaatac
     1981 gcaaggggaa cgcatgaagg ttatcgctgt acttaaccag aaaggcgggt caggcaagac
     2041 gaccatcgca 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 ctgaatacat cgcgcagcta ccagagtaaa tgagcaaatg aataaatgag
     2881 tagatgaatt ttagcggcta aaggaggcgg catggaaaat caagaacaac caggcaccga
     2941 cgccgtggaa tgccccatgt gtggaggaac gggcggttgg ccaggcgtaa gcggctgggt
     3001 tgcctgccgg ccctgcaatg gcactggaac ccccaagccc gaggaatcgg cgtgagcggt
     3061 cgcaaaccat ccggcccggt acaaatcggc gcggcgctgg gtgatgacct ggtggagaag
     3121 ttgaaggccg cgcaggccgc ccagcggcaa cgcatcgagg cagaagcacg ccccggtgaa
     3181 tcgtggcaag cggccgctga tcgaatccgc aaagaatccc ggcaaccgcc ggcagccggt
     3241 gcgccgtcga ttaggaagcc gcccaagggc gacgagcaac cagatttttt cgttccgatg
     3301 ctctatgacg tgggcacccg cgatagtcgc agcatcatgg acgtggccgt tttccgtctg
     3361 tcgaagcgtg accgacgagc tggcgaggtg atccgctacg agcttccaga cgggcacgta
     3421 gaggtttccg cagggccggc cggcatggcc agtgtgtggg attacgacct ggtactgatg
     3481 gcggtttccc atctaaccga atccatgaac cgataccggg aagggaaggg agacaagccc
     3541 ggccgcgtgt tccgtccaca cgttgcggac gtactcaagt tctgccggcg agccgatggc
     3601 ggaaagcaga aagacgacct ggtagaaacc tgcattcggt taaacaccac gcacgttgcc
     3661 atgcagcgta cgaagaaggc caagaacggc cgcctggtga cggtatccga gggtgaagcc
     3721 ttgattagcc gctacaagat cgtaaagagc gaaaccgggc ggccggagta catcgagatc
     3781 gagctagctg attggatgta ccgcgagatc acagaaggca agaacccgga cgtgctgacg
     3841 gttcaccccg attacttttt gatcgatccc ggcatcggcc gttttctcta ccgcctggca
     3901 cgccgcgccg caggcaaggc agaagccaga tggttgttca agacgatcta cgaacgcagt
     3961 ggcagcgccg gagagttcaa gaagttctgt ttcaccgtgc gcaagctgat cgggtcaaat
     4021 gacctgccgg agtacgattt gaaggaggag gcggggcagg ctggcccgat cctagtcatg
     4081 cgctaccgca acctgatcga gggcgaagca tccgccggtt cctaatgtac ggagcagatg
     4141 ctagggcaaa ttgccctagc aggggaaaaa ggtcgaaaag gactctttcc tgtggatagc
     4201 acgtacattg ggaacccaaa gccgtacatt gggaaccgga acccgtacat tgggaaccca
     4261 aagccgtaca ttgggaaccg gtcacacatg taagtgactg atataaaaga gaaaaaaggc
     4321 gatttttccg cctaaaactc tttaaaactt attaaaactc ttaaaacccg cctggcctgt
     4381 gcataactgt ctggccagcg cacagccgaa gagctgcaaa aagcgcctac ccttcggtcg
     4441 ctgcgctccc tacgccccgc cgcttcgcgt cggcctatcg cggccgctgg ccgctcaaaa
     4501 atggctggcc tacggccagg caatctacca gggcgcggac aagccgcgcc gtcgccactc
     4561 gaccgccggc gcccacatca aggcaccctg cctcgcgcgt ttcggtgatg acggtgaaaa
     4621 cctctgacac atgcagctcc cggtgacggt cacagcttgt ctgtaagcgg atgccgggag
     4681 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggcg cagccatgac
     4741 ccagtcacgt agcgatagcg gagtgtatac tggcttaact atgcggcatc agagcagatt
     4801 gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac
     4861 cgcatcaggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     4921 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     4981 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     5041 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     5101 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     5161 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     5221 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     5281 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     5341 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     5401 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     5461 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
     5521 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     5581 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     5641 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     5701 taagggattt tggtcatgca ttctaggtga ttagaaaaac tcatcgagca tcaaatgaaa
     5761 ctgcaattta ttcatatcag gattatcaat accatatttt tgaaaaagcc gtttctgtaa
     5821 tgaaggagaa aactcaccga ggcagttcca taggatggca agatcctggt atcggtctgc
     5881 gattccgact cgtccaacat caatacaacc tattaatttc ccctcgtcaa aaataaggtt
     5941 atcaagtgag aaatcaccat gagtgacgac tgaatccggt gagaatggca aaagtttatg
     6001 catttctttc cagacttgtt caacaggcca gccattacgc tcgtcatcaa aatcactcgc
     6061 atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg agtcgaaata cgcgatcgct
     6121 gttaaaagga caattacaaa caggaatcga atgcaaccgg cgcaggaaca ctgccagcgc
     6181 atcaacaata ttttcacctg aatcaggata ttcttctaat acctggaatg ctgttttccc
     6241 tgggatcgca gtggtgagta accatgcatc atcaggagta cggataaaat gcttgatggt
     6301 cggaagaggc ataaattccg tcagccagtt tagtctgacc atctcatctg taacatcatt
     6361 ggcaacgcta cctttgccat gtttcagaaa caactctggc gcatcgggct tcccatacaa
     6421 tcggtagatt gtcgcacctg attgcccgac attatcgcga gcccatttat acccatataa
     6481 atcagcatcc atgttggaat ttaatcgcgg ccttgagcaa gacgtttccc gttgaatatg
     6541 gctcataaca gaacttatta tttccttcct cttttctaca gtatttaaag ataccccaag
     6601 aagctaatta taacaagacg aactccaatt cactgttcct tgcattctaa aaccttaaat
     6661 accagaaaac agctttttca aagttgtttt caaagttggc gtataacata gtatcgacgg
     6721 agccgatttt gaaaccgcgg tgatcacagg cagcaacgct ctgtcatcgt tacaatcaac
     6781 atgctaccct ccgcgagatc atccgtgttt caaacccggc agcttagttg ccgttcttcc
     6841 gaatagcatc ggtaacatga gcaaagtctg ccgccttaca acggctctcc cgctgacgcc
     6901 gtcccggact gatgggctgc ctgtatcgag tggtgatttt gtgccgagct gccggtcggg
     6961 gagctgttgg ctggctggtg gcaggatata ttgtggtgta aacataacga attcgtctca
//
