LOCUS       sequence-134268-        6188 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fusarium graminearum AurZ encoded on CEN/ARS yeast shuttle vector 
            with DS2 selection marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6188)
  AUTHORS   Rugbjerg P, Myling-Petersen N, Sommer MO
  TITLE     Flexible metabolic pathway construction using modular and divisible 
            selection gene regulators.
  JOURNAL   Metab Eng. 2015 Sep;31:189-97. doi: 10.1016/j.ymben.2015.08.004. 
            Epub 2015 Aug 21.
  PUBMED    26303342
REFERENCE   2  (bases 1 to 6188)
  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: "Metab 
            Eng."; date: "2015-09"; volume: "31:189-97. doi"; pages: " 
            10.1016/j.ymben.2015.08.004. Epub 2015 Aug 21"
COMMENT     Sequence Label: pDS2-aurZ
FEATURES             Location/Qualifiers
     source          1..6188
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     234..255
                     /label=LexA
                     /note="LexA DNA binding domain, forward primer"
     protein_bind    309..333
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    1178..1211
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        complement(2217..2616)
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     primer_bind     complement(2737..2754)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2908..3496)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2988..3007)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3667..4458,4459..4527))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4290..4309
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4528..4632)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    4669..5172
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     5214..5232
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5270..5292)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     protein_bind    5413..5446
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             5852..5872
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(5879..6130,6131..6188,1..296)
                     /codon_start=1
                     /gene="lexA"
                     /product="E. coli DNA-binding protein that represses genes 
                     involved in the SOS response to DNA damage"
                     /label=LexA
                     /note="The N-terminal DNA-binding domain recognizes the 
                     16-bp palindromic sequence 5'-CTGTATATATATACAG-3'."
                     /translation="MKALTARQQEVFDLIRDHISQTGMPPTRAEIAQRLGFRSPNAAEE
                     HLKALARKGVIEIVSGASRGIRLLQEEEEGLPLVGRVAAGEPLLAQQHIEGHYQVDPSL
                     FKPNADFLLRVSGMSMKDIGIMDGDLLAVHKTQDVRNGQVVVARIDDEVTVKRLKKQGN
                     KVELLPENSEFKPIVVDLRQQSFTIEGLAVGVIRNGDWL"
ORIGIN
        1 tattcaagcc gaatgctgat ttcctgctgc gcgtcagcgg gatgtcgatg aaagatatcg
       61 gcattatgga tggtgacttg ctggcagtgc ataaaactca ggatgtacgt aacggtcagg
      121 tcgttgtcgc acgtattgat gacgaagtta ccgttaagcg cctgaaaaaa cagggcaata
      181 aagtcgaact gttgccagaa aatagcgagt ttaaaccaat tgtcgttgac cttcgtcagc
      241 agagcttcac cattgaaggg ctggcggttg gggttattcg caacggcgac tggctgaggt
      301 cgaatcaaac aagtttgtac aaaaaagcag gctccgcggc cgcccccttc acccgtgagt
      361 acaagctagt ggtccttggt tcaggaggcg ttgggaagtc tgctctgaca gttcagtttg
      421 ttcagggaat ttttgttgaa aaatatgacc caacgataga agattcctac agaaagcaag
      481 ttgaagtcga ttgccaacag tgtatgctcg aaatcctgga tactgcaggg acagagcaat
      541 ttacagcaat gagggatttg tatatgaaga acggccaagg ttttgcacta gtatattcta
      601 ttacagctca gtccacgttt aacgacttac aggacctgag ggaacagatt ttacgggtta
      661 aggacacgga agatgttcca atgattttgg ttggcaataa atgtgacctg gaagatgagc
      721 gagtagttgg caaagagcag ggccagaatt tagcaagaca gtggtgtaac tgtgcctttt
      781 tagaatcttc tgcaaagtca aagatcaatg ttaatgagat attttatgac ctggtcagac
      841 agataaatag gaaaacgcca gtggaaaaga agaagcctaa aaagaaatca tgtctgctgc
      901 tctagaagcg gtaactcgag gacaataaaa agattcttgt tttcaagaac ttgtcatttg
      961 tatagttttt ttatattgta gttgttctat tttaatcaaa tgttagcgtg atttatattt
     1021 tttttcgcct cgacatcatc tgcccagatg cgaagttaag tgcgcagaaa gtaatatcat
     1081 gcgtcaatcg tatgtgaatg ctggtcgcta tactgctgtc gattcgatac taacgccgcc
     1141 atccagtgtc gaaaacgagc tctcgagaac ccttaatata acttcgtata atgtatgcta
     1201 tacgaagtta ttaggtgata tcagatccac tagtggccta tgcacccaat tgcgcaactg
     1261 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctgattt gcccgggcag
     1321 ttcaggctca tcaggcgcgc catgcaggat gcattgatca gttaacccat gggcatgcat
     1381 ttccttccat cttgtgattc atgctatcca tcttttttga gtatccaatt aacgaagacg
     1441 ttaccagctg attgaaggtt ctcaaagtga ctgtactcca tgttttctta tcatccatgt
     1501 agttattttt caaactgcaa attcaagaaa aagccacgcg tgtgcacctt ttttttcccc
     1561 ttccagtgca ttatgcaata gacagcacga gtctttgaaa aagtaactta taaaactgta
     1621 tcaattttta aacctaaata gattcataaa ctattcgtta atataaagtg ttctaaacta
     1681 tgatgaaaaa ataagcagaa aagactaata attcttagtt aaaagcactc cgcggttaat
     1741 cttttggaac cactttgcca tcgttaacga actcttcata gtatccaact gtcatggttg
     1801 atcttttagt atctgcgaaa ttaacatggt ctggcgccac tttttctaaa aaccttggat
     1861 cagctttcat tctcaggaag tcctttatat ccttaaagat aaactggacg atacaatcgt
     1921 aatcagattg attggaaaat tgaggatcat atagttggta caacattggt ctagtttcct
     1981 tagtgttgtg ggtcattgtg taacggacaa taccatactc ttccatgaga ggtgatacca
     2041 attttgcgtg aacttttgtc atgtgatgtc tgtattcctt ttcgctaaga gatggctttt
     2101 tgtaggcagt aattgtcaag cagactaaag tgtctggttg acctgttaca gtagcttcat
     2161 gagatgtctc ttgcgtgtct gaatgaggac tcattttaag ctttttgtaa ttaaaactta
     2221 gattagattg ctatgctttc tttctaatga gcaagaagta aaaaaagttg taatagaaca
     2281 agaaaaatga aactgaaact tgagaaattg aagaccgttt attaacttaa atatcaatgg
     2341 gaggtcatcg aaagagaaaa aaatcaaaaa aaaaaatttt caagaaaaag aaacgtgata
     2401 aaaattttta ttgccttttt cgacgaagaa aaagaaacga ggcggtctct tttttctttt
     2461 ccaaaccttt agtacgggta attaacgaca ccctagagga agaaagaggg gaaatttagt
     2521 atgctgtgct tgggtgtttt gaagtggtac ggcgatgcgc ggagtccgag aaaatctgga
     2581 agagtaaaaa aggagtagaa acattttgaa gctaagatct gatatccctg catggcgcgc
     2641 ctgatgagcc tgaactgccc gggcaaatca gctgcattaa tgaatcggcc aacgcgcggg
     2701 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     2761 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     2821 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     2881 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     2941 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     3001 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     3061 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     3121 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     3181 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     3241 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     3301 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     3361 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     3421 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     3481 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     3541 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     3601 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     3661 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     3721 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     3781 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     3841 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     3901 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     3961 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     4021 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     4081 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     4141 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     4201 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     4261 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     4321 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     4381 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     4441 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     4501 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     4561 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     4621 aggggttccg cgcacatttc cccgaaaagt gccacctggg tccttttcat cacgtgctat
     4681 aaaaataatt ataatttaaa ttttttaata taaatatata aattaaaaat agaaagtaaa
     4741 aaaagaaatt aaagaaaaaa tagtttttgt tttccgaaga tgtaaaagac tctaggggga
     4801 tcgccaacaa atactacctt ttatcttgct cttcctgctc tcaggtatta atgccgaatt
     4861 gtttcatctt gtctgtgtag aagaccacac acgaaaatcc tgtgatttta cattttactt
     4921 atcgttaatc gaatgtatat ctatttaatc tgcttttctt gtctaataaa tatatatgta
     4981 aagtacgctt tttgttgaaa ttttttaaac ctttgtttat ttttttttct tcattccgta
     5041 actcttctac cttctttatt tactttctaa aatccaaata caaaacataa aaataaataa
     5101 acacagagta aattcccaaa ttattccatc attaaaagat acgaggcgcg tgtaagttac
     5161 aggcaagcga tccgtcctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     5221 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     5281 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     5341 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat acagctttcg
     5401 acaaccctta atataacttc gtataatgta tgctatacga agttattagg tctagagatc
     5461 tgtttagctt gcctcgtccc cgccgggtca cccggccagc gacatggagg cccagaatac
     5521 cctccttgac agtcttgacg tgcgcagctc aggggcatga tgtgactgtc gcccgtacat
     5581 ttagcccata catccccatg tataatcatt tgcatccata cattttgatg gccgcacggc
     5641 gcgaagcaaa aattacggct cctcgctgca gacctgcgag cagggaaacg ctcccctcac
     5701 agacgcgttg aattgtcccc acgccgcgcc cctgtagaga aatataaaag gttaggattt
     5761 gccactgagg ttcttctttc atatacttcc ttttaaaatc ttgctaggat acagttctca
     5821 catcacatcc gaacataaac aagaacatat gcccaagaag aagcggaagg tctcgagcat
     5881 gaaagcgtta acggccaggc aacaagaggt gtttgatctc atccgtgatc acatcagcca
     5941 gacaggtatg ccgccgacgc gtgcggaaat cgcgcagcgt ttggggttcc gttccccaaa
     6001 cgcggctgaa gaacatctga aggcgctggc acgcaaaggc gttattgaaa ttgtttccgg
     6061 cgcatcacgc gggattcgtc tgttgcagga agaggaagaa gggttgccgc tggtaggtcg
     6121 tgtggctgcc ggtgaaccac ttctggcgca acagcatatt gaaggtcatt atcaggtcga
     6181 tccttcct
//
