LOCUS       sequence-336986-        4544 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A kanMX-marked pFA6a based plasmid, which has a 7-aa linker, a 16-aa
            XTEN16 linker, three tandam repeats of sAID. It serves as the 
            template for 3_sAID degron tagging at the C-terminus..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4544)
  AUTHORS   Zhang XR, Zhao L, Suo F, Gao Y, Wu Q, Qi X, Du LL
  TITLE     An improved auxin-inducible degron system for fission yeast.
  JOURNAL   G3 (Bethesda). 2022 Jan 4;12(1):jkab393. doi: 
            10.1093/g3journal/jkab393.
  PUBMED    34849776
REFERENCE   2  (bases 1 to 4544)
  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: "G3 
            (Bethesda)."; date: "2022-01-4"; volume: "12(1):jkab393. doi"; 
            pages: " 10.1093/g3journal/jkab393"
COMMENT     Sequence Label: pDB4581
FEATURES             Location/Qualifiers
     source          1..4544
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        7..25
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     7..24
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     terminator      482..669
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(537..561)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            744..2100
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        744..1087
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1088..1897
                     /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(1155..1174)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      1903..2100
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2204..2223)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2205..2223)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2310..2327)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2481..3069)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2561..2580)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3240..4031,4032..4100))
                     /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     3863..3882
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4101..4205)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4273..4291
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4329..4351)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4451..4470
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 catacgattt aggtgacact atagaacgcg gccgccagct gaagcttcgt acgctgcagg
       61 tcgaccggat ccccgggtta attaactctg gttctgaaac tcctggtact tctgagtctg
      121 ctactcccga atctcctaag gatcctgcca agcctccggc taaagctcag gttgtcggct
      181 ggcctccagt tcgctcttat cgtaagaacg taatggtctc ttgccaaaag tcttccggtg
      241 gacccaagga ccctgctaag cccccagcca aggcccaagt tgtaggatgg ccccctgttc
      301 gttcctatcg caagaatgtt atggttagct gtcagaagtc ctctggaggt cctaaagatc
      361 ccgccaaacc tcccgctaag gctcaagtcg ttggttggcc acctgtccgt tcttaccgta
      421 aaaatgtcat ggtttcttgt caaaaatctt caggcggtta aggcgcgcca cttctaaata
      481 agcgaatttc ttatgattta tgatttttat tattaaataa gttataaaaa aaataagtgt
      541 atacaaattt taaagtgact cttaggtttt aaaacgaaaa ttcttattct tgagtaactc
      601 tttcctgtag gtcaggttgc tttctcaggt atagtatgag gtcgctctta ttgaccacac
      661 ctctaccggc agatccgcta gggataacag ggtaatatag atctgtttag cttgcctcgt
      721 ccccgccggg tcacccggcc agcgacatgg aggcccagaa taccctcctt gacagtcttg
      781 acgtgcgcag ctcaggggca tgatgtgact gtcgcccgta catttagccc atacatcccc
      841 atgtataatc atttgcatcc atacattttg atggccgcac ggcgcgaagc aaaaattacg
      901 gctcctcgct gcagacctgc gagcagggaa acgctcccct cacagacgcg ttgaattgtc
      961 cccacgccgc gcccctgtag agaaatataa aaggttagga tttgccactg aggttcttct
     1021 ttcatatact tccttttaaa atcttgctag gatacagttc tcacatcaca tccgaacata
     1081 aacaaccatg ggtaaggaaa agactcacgt ttcgaggccg cgattaaatt ccaacatgga
     1141 tgctgattta tatgggtata aatgggctcg cgataatgtc gggcaatcag gtgcgacaat
     1201 ctatcgattg tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag
     1261 cgttgccaat gatgttacag atgagatggt cagactaaac tggctgacgg aatttatgcc
     1321 tcttccgacc atcaagcatt ttatccgtac tcctgatgat gcatggttac tcaccactgc
     1381 gatccccggc aaaacagcat tccaggtatt agaagaatat cctgattcag gtgaaaatat
     1441 tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg attcctgttt gtaattgtcc
     1501 ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt
     1561 ggttgatgcg agtgattttg atgacgagcg taatggctgg cctgttgaac aagtctggaa
     1621 agaaatgcat aagcttttgc cattctcacc ggattcagtc gtcactcatg gtgatttctc
     1681 acttgataac cttatttttg acgaggggaa attaataggt tgtattgatg ttggacgagt
     1741 cggaatcgca gaccgatacc aggatcttgc catcctatgg aactgcctcg gtgagttttc
     1801 tccttcatta cagaaacggc tttttcaaaa atatggtatt gataatcctg atatgaataa
     1861 attgcagttt catttgatgc tcgatgagtt tttctaatca gtactgacaa taaaaagatt
     1921 cttgttttca agaacttgtc atttgtatag tttttttata ttgtagttgt tctattttaa
     1981 tcaaatgtta gcgtgattta tatttttttt cgcctcgaca tcatctgccc agatgcgaag
     2041 ttaagtgcgc agaaagtaat atcatgcgtc aatcgtatgt gaatgctggt cgctatactg
     2101 ctgtcgattc gatactaacg ccgccatcca gtttaaacga gctcgaattc atcgatgata
     2161 tcagatccac tagtggccta tgcggccgcg gatctgccgg tctccctata gtgagtcgta
     2221 ttaatttcga taagccaggt taacctgcat taatgaatcg gccaacgcgc ggggagaggc
     2281 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     2341 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     2401 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     2461 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     2521 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     2581 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     2641 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     2701 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     2761 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     2821 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     2881 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     2941 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     3001 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     3061 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     3121 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     3181 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     3241 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     3301 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     3361 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     3421 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     3481 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     3541 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     3601 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     3661 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     3721 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     3781 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     3841 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     3901 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     3961 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     4021 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     4081 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     4141 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     4201 ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca
     4261 ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac
     4321 ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat
     4381 gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg
     4441 cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgga catattgtcg
     4501 ttagaacgcg gctacaatta atacataacc ttatgtatca taca
//
