LOCUS       sequence-182942-        4654 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  KanMX Z3EV promoter cassette.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4654)
  AUTHORS   Ohira MJ, Hendrickson DG, Scott McIsaac R, Rhind N
  TITLE     An estradiol-inducible promoter enables fast, graduated control of 
            gene expression in fission yeast.
  JOURNAL   Yeast. 2017 Aug;34(8):323-334. doi: 10.1002/yea.3235. Epub 2017 Jun 
            8.
  PUBMED    28423198
REFERENCE   2  (bases 1 to 4654)
  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"; doi: "10.1002/yea.3235"; 
            journalName: "Yeast"; date: "2017-08"; volume: "34"; issue: "8"; 
            pages: "323-334"
COMMENT     Sequence Label: pFS478
FEATURES             Location/Qualifiers
     source          1..4654
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     610..633
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     promoter        complement(726..744)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(727..744)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(825..844)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     944..966
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1004..1022)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1090..1194
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1195..1263,1264..2055)
                     /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     complement(1413..1432)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2226..2814
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2715..2734
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2968..2985
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     3072..3091
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3072..3090
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     gene            complement(3195..4551)
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     terminator      complement(3195..3392)
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             complement(3398..4207)
                     /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     4121..4140
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(4208..4551)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
ORIGIN
        1 gaagtttaat aatcatatta catggcatta ccaccatata catatccata tacatatcca
       61 tatctaatct tacttatatg ttgtggaaat gtaaagagcc ccattatctt agcctaaaaa
      121 aaccttctct ttggaacttt cagtaatacg cttaactgct cattgctata ttgaagtgcg
      181 gccgcgtggg cgtgcgtggg cgggcgtggg cgtgcgtggg cgggcgtggg cgtgcgtggg
      241 cgtctagacc gtgcgtcctc gtcttcaccg gtcgcgttcc tgaaacgcag atgtgcctcg
      301 cgccgcactg ctccgaacaa taaagattct acaatactag cttttatggt tatgaagagg
      361 aaaaattggc agtaacctgg ccccacaaac cttcaaatta acgaatcaaa ttaacaacca
      421 taggatgata atgcgattag ttttttagcc ttatttctgg ggtaattaat cagcgaagcg
      481 atgatttttg atctattaac agatatataa atggaaaagc tgcataacca ctttaactaa
      541 tactttcaac attttcagtt tgtattactt cttattcaaa tgtcataaaa gtatcaacaa
      601 aaaattgtta atatacctct atactttaac gtcaaggaga aaaaactata ttgttaaatc
      661 atgtctttaa ttaacccggg gatccgtcga cctgcagcgt acgaagcttc agctggcggc
      721 cgcgttctat agtgtcacct aaatcgtatg tgtatgatac ataaggttat gtattaattg
      781 tagccgcgtt ctaacgacaa tatgtccata tggtgcactc tcagtacaat ctgctctgat
      841 gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
      901 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
      961 cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta
     1021 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     1081 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     1141 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     1201 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     1261 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     1321 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     1381 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     1441 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     1501 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     1561 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     1621 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     1681 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     1741 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     1801 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     1861 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     1921 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     1981 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     2041 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     2101 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     2161 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     2221 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     2281 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     2341 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     2401 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     2461 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     2521 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     2581 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     2641 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     2701 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     2761 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     2821 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     2881 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     2941 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     3001 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag gttaacctgg
     3061 cttatcgaaa ttaatacgac tcactatagg gagaccggca gatccgcggc cgcataggcc
     3121 actagtggat ctgatatcat cgatgaattc gagctcgttt aaactggatg gcggcgttag
     3181 tatcgaatcg acagcagtat agcgaccagc attcacatac gattgacgca tgatattact
     3241 ttctgcgcac ttaacttcgc atctgggcag atgatgtcga ggcgaaaaaa aatataaatc
     3301 acgctaacat ttgattaaaa tagaacaact acaatataaa aaaactatac aaatgacaag
     3361 ttcttgaaaa caagaatctt tttattgtca gtactgatta gaaaaactca tcgagcatca
     3421 aatgaaactg caatttattc atatcaggat tatcaatacc atatttttga aaaagccgtt
     3481 tctgtaatga aggagaaaac tcaccgaggc agttccatag gatggcaaga tcctggtatc
     3541 ggtctgcgat tccgactcgt ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa
     3601 taaggttatc aagtgagaaa tcaccatgag tgacgactga atccggtgag aatggcaaaa
     3661 gcttatgcat ttctttccag acttgttcaa caggccagcc attacgctcg tcatcaaaat
     3721 cactcgcatc aaccaaaccg ttattcattc gtgattgcgc ctgagcgaga cgaaatacgc
     3781 gatcgctgtt aaaaggacaa ttacaaacag gaatcgaatg caaccggcgc aggaacactg
     3841 ccagcgcatc aacaatattt tcacctgaat caggatattc ttctaatacc tggaatgctg
     3901 ttttgccggg gatcgcagtg gtgagtaacc atgcatcatc aggagtacgg ataaaatgct
     3961 tgatggtcgg aagaggcata aattccgtca gccagtttag tctgaccatc tcatctgtaa
     4021 catcattggc aacgctacct ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc
     4081 catacaatcg atagattgtc gcacctgatt gcccgacatt atcgcgagcc catttatacc
     4141 catataaatc agcatccatg ttggaattta atcgcggcct cgaaacgtga gtcttttcct
     4201 tacccatggt tgtttatgtt cggatgtgat gtgagaactg tatcctagca agattttaaa
     4261 aggaagtata tgaaagaaga acctcagtgg caaatcctaa ccttttatat ttctctacag
     4321 gggcgcggcg tggggacaat tcaacgcgtc tgtgagggga gcgtttccct gctcgcaggt
     4381 ctgcagcgag gagccgtaat ttttgcttcg cgccgtgcgg ccatcaaaat gtatggatgc
     4441 aaatgattat acatggggat gtatgggcta aatgtacggg cgacagtcac atcatgcccc
     4501 tgagctgcgc acgtcaagac tgtcaaggag ggtattctgg gcctccatgt cgctggccgg
     4561 gtgacccggc ggggacgagg caagctaaac agatctcgcc ataaattata ttgaattttc
     4621 aaaaattctt actttttttt tggatggacg caaa
//
