LOCUS       sequence-252436-        5595 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Z. rouxii episomal vector containing G418 resistance.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5595)
  AUTHORS   Bizzarri M, Cassanelli S, Duskova M, Sychrova H, Solieri L
  TITLE     A set of plasmids carrying antibiotic resistance markers and Cre 
            recombinase for genetic engineering of nonconventional yeast 
            Zygosaccharomyces rouxii.
  JOURNAL   Yeast. 2019 Dec;36(12):711-722. doi: 10.1002/yea.3438. Epub 2019 Sep
            11.
  PUBMED    31414502
REFERENCE   2  (bases 1 to 5595)
  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"; doi: "10.1002/yea.3438"; 
            journalName: "Yeast"; date: "2019-12"; volume: "36"; issue: "12"; 
            pages: "711-722"
COMMENT     Sequence Label: pZEG
FEATURES             Location/Qualifiers
     source          1..5595
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(146..469)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     378..395
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    396..452
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(465..481)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(465..481)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(478..500)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    489..505
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(513..519,520..537,538..543))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    558..579
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     protein_bind    734..767
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     gene            822..2178
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        822..1165
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1166..1975
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1233..1252)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      1981..2178
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     protein_bind    2241..2274
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        complement(2307..2518)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(2596..2613)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3776..4364)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3856..3875)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4535..5326,5327..5395))
                     /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     5158..5177
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5396..5500)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5568..5586
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt cgagctcggt acccggggat
      421 cctctagagt cgacctgcag gcatgcaagc ttggcgtaat catggtcata gctgtttcct
      481 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
      541 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
      601 gctttccagt cgggaaacct gtcgtgccag cagatctggc tttttctttc caattttttt
      661 tttgtgagtt tagtatacat gcatttactt ataatacagt tttttcgtac gctgcaggtc
      721 gacaaccctt aatataactt cgtataatgt atgctatacg aagttattag gtctagagat
      781 ctgtttagct tgcctcgtcc ccgccgggtc acccggccag cgacatggag gcccagaata
      841 ccctccttga cagtcttgac gtgcgcagct caggggcatg atgtgactgt cgcccgtaca
      901 tttagcccat acatccccat gtataatcat ttgcatccat acattttgat ggccgcacgg
      961 cgcgaagcaa aaattacggc tcctcgctgc agacctgcga gcagggaaac gctcccctca
     1021 cagacgcgtt gaattgtccc cacgccgcgc ccctgtagag aaatataaaa ggttaggatt
     1081 tgccactgag gttcttcttt catatacttc cttttaaaat cttgctagga tacagttctc
     1141 acatcacatc cgaacataaa caaccatggg taaggaaaag actcacgttt cgaggccgcg
     1201 attaaattcc aacatggatg ctgatttata tgggtataaa tgggctcgcg ataatgtcgg
     1261 gcaatcaggt gcgacaatct atcgattgta tgggaagccc gatgcgccag agttgtttct
     1321 gaaacatggc aaaggtagcg ttgccaatga tgttacagat gagatggtca gactaaactg
     1381 gctgacggaa tttatgcctc ttccgaccat caagcatttt atccgtactc ctgatgatgc
     1441 atggttactc accactgcga tccccggcaa aacagcattc caggtattag aagaatatcc
     1501 tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat
     1561 tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc
     1621 acgaatgaat aacggtttgg ttgatgcgag tgattttgat gacgagcgta atggctggcc
     1681 tgttgaacaa gtctggaaag aaatgcataa gcttttgcca ttctcaccgg attcagtcgt
     1741 cactcatggt gatttctcac ttgataacct tatttttgac gaggggaaat taataggttg
     1801 tattgatgtt ggacgagtcg gaatcgcaga ccgataccag gatcttgcca tcctatggaa
     1861 ctgcctcggt gagttttctc cttcattaca gaaacggctt tttcaaaaat atggtattga
     1921 taatcctgat atgaataaat tgcagtttca tttgatgctc gatgagtttt tctaatcagt
     1981 actgacaata aaaagattct tgttttcaag aacttgtcat ttgtatagtt tttttatatt
     2041 gtagttgttc tattttaatc aaatgttagc gtgatttata ttttttttcg cctcgacatc
     2101 atctgcccag atgcgaagtt aagtgcgcag aaagtaatat catgcgtcaa tcgtatgtga
     2161 atgctggtcg ctatactgct gtcgattcga tactaacgcc gccatccagt gtcgaaaacg
     2221 agctctcgag aacccttaat ataacttcgt ataatgtatg ctatacgaag ttattaggtg
     2281 atatcagatc cactagtggc ctatgcgatt tatcttcgtt tccggttttt gttctgtgca
     2341 gttgggttaa gaatactggg caatttcatg tttcttcaac actacatatg cgtatatata
     2401 ccaatctaag tctgtgctcc ttccttcgtt cttccttctg ttcggagatt accgaatcaa
     2461 aaaaatttca aggaaaccga aatcaaaaaa aagaataaaa aaaaaatgat gaattgaaaa
     2521 gccagatctg ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     2581 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     2641 ggtatcagca tcgatagatc ctacaaaata tgtaatgcta ttcttagatt cccacgagag
     2701 gcttgctttt gtacatagac cctccagaaa acatagtgct gtttatgcag cacacagaag
     2761 gtttataggt gtgagctaca ctttataggc cgggtaagta tagtaagata tgtgcaaatt
     2821 agccgcggag ccataaacaa tcatctttta aattagatgg cacagcctta aaccaagcaa
     2881 ttctggtcac ctctccggat tttgggtgaa tatcaaaatg acctcacaag gtaacacttt
     2941 ggatgatatt cggttacaag tcaggtgtga cattctaggt aaagacacca acagaagagg
     3001 aggctctgag aaggatcctg aggatcaaag tgaaactgag tccagtggat agactcattt
     3061 gctaatttaa gcctttctgt aagactattg agaccgtgta ctattaattt ttcttattgt
     3121 aacagtaggt aataggtaag ggtataaaaa tcatgggtat aatgataaat ttgtattgca
     3181 ataatgtcca aatcaaattc ggaggcgagg tccaagaagc aagatggtgt atggtagcaa
     3241 tatagagagg gttttattgg gttattattt tattttcaag aatttacatt ttgagtgtag
     3301 atctttactt tcagcaagaa ggtacgagga acaagcgggt agtttagagg aacaaaataa
     3361 aaatacaaga gcatagtaac tgtcagggtg tatcaactgg accaatcatt gcttctcgta
     3421 gtcaaatgtt tattcgatag agtaatggtt taatttctgt agcaaccgcc tgcgcaggct
     3481 cccttctgcg cggtacgggt tgatgaaaga acgacttgat gaaagaacga tttgatgaaa
     3541 gaacgaatta atgaaagaac gatttgatga aagaataacg tattctttca tcaacgtcca
     3601 cagtgatatg atctcgccta gaaaaactaa attatattaa gctaaactaa atcgatgctc
     3661 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     3721 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     3781 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     3841 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     3901 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     3961 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     4021 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     4081 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     4141 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     4201 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     4261 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     4321 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     4381 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     4441 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     4501 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     4561 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     4621 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     4681 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     4741 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     4801 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     4861 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     4921 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     4981 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     5041 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     5101 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     5161 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     5221 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     5281 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     5341 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     5401 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     5461 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     5521 cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca
     5581 cgaggccctt tcgtc
//
