LOCUS       sequence-475897-        9750 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express the lower portion of melvaonate pathway enzymes (Erg12p, 
            Erg8p, and Erg19p) in the peroxisome; chromosomal integration by 
            CRISPR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9750)
  TITLE     Dueber Lab plasmids 2025
REFERENCE   2  (bases 1 to 9750)
  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"
COMMENT     Sequence Label: pKS1461
FEATURES             Location/Qualifiers
     source          1..9750
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      2110..2297
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2165..2189)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             5709..5720
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             complement(7488..8303)
                     /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(7488..8303)
                     /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     8211..8230
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     8295..8314
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      8528..9116
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9017..9036
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 actggccgat aattgcagac gaacgcaccc atgaaccaca cggttagtcc aaaaggggca
       61 gttcagattc cagatgcggg aattagcttg ctgccaccct cacctcacta acgctgcggt
      121 gtgcggatac ttcatgctat ttatagacgc gcgtgtcgga atcagcacgc gcaagaacca
      181 aatgggaaaa tcggaatggg tccagaactg ctttgagtgc tggctattgg cgtctgattt
      241 ccgttttggg aatcctttgc cgcgcgcccc tctcaaaact ccgcacaagt cccagaaagc
      301 gggaaagaaa taaaacgcca ccaaaaaaaa aaaaataaaa gccaatcctc gaagcgtggg
      361 tggtaggccc tggattatcc cgtacaagta tttctcagga gtaaaaaaac cgtttgtttt
      421 ggaatttccc atttcgcggc cacctacgcc gctatctttg caacaactat ctgcgataac
      481 tcagcaaatt ttgcatattc gtgttgcagt attgcgataa tgggagtctt acttccaaca
      541 taacggcaga aagaaatgtg agaaaatttt gcatcctttg cctccgttca agtatataaa
      601 gtcggcatgc ttgataatct ttctttccat cctacattgt tctaattatt cttattctcc
      661 tttattcttt cctaacatac caagaaatta atcttctgtc attcgcttaa acactatatc
      721 aataaagatc tatgtcatta ccgttcttaa cttctgcacc gggaaaggtt attatttttg
      781 gtgaacactc tgctgtgtac aacaagcctg ccgtcgctgc tagtgtgtct gcgttgagaa
      841 cctacctgct aataagcgag tcatctgcac cagatactat tgaattggac ttcccggaca
      901 ttagctttaa tcataagtgg tccatcaatg atttcaatgc catcaccgag gatcaagtaa
      961 actcccaaaa attggccaag gctcaacaag ccaccgatgg cttgtctcag gaactcgtta
     1021 gtcttttgga tccgttgtta gctcaactat ccgaatcctt ccactaccat gcagcgtttt
     1081 gtttcctgta tatgtttgtt tgcctatgcc cccatgccaa gaatattaag ttttctttaa
     1141 agtctacttt acccatcggt gctgggttgg gctcaagcgc ctctatttct gtatcactgg
     1201 ccttagctat ggcctacttg ggggggttaa taggatctaa tgacttggaa aagctgtcag
     1261 aaaacgataa gcatatagtg aatcaatggg ccttcatagg tgaaaagtgt attcacggta
     1321 ccccttcagg aatagataac gctgtggcca cttatggtaa tgccctgcta tttgaaaaag
     1381 actcacataa tggaacaata aacacaaaca attttaagtt cttagatgat ttcccagcca
     1441 ttccaatgat cctaacctat actagaattc caaggtctac aaaagatctt gttgctcgcg
     1501 ttcgtgtgtt ggtcaccgag aaatttcctg aagttatgaa gccaattcta gatgccatgg
     1561 gtgaatgtgc cctacaaggc ttagagatca tgactaagtt aagtaaatgt aaaggcaccg
     1621 atgacgaggc tgtagaaact aataatgaac tgtatgaaca actattggaa ttgataagaa
     1681 taaatcatgg actgcttgtc tcaatcggtg tttctcatcc tggattagaa cttattaaaa
     1741 atctgagcga tgatttgaga attggctcca caaaacttac cggtgctggt ggcggcggtt
     1801 gctctttgac tttgttacga agagacatta ctcaagagca aattgacagc ttcaaaaaga
     1861 aattgcaaga tgattttagt tacgagacat ttgaaacaga cttgggtggg actggctgct
     1921 gtttgttaag cgcaaaaaat ttgaataaag atcttaaaat caaatcccta gtattccaat
     1981 tatttgaaaa taaaactacc acaaagcaac aaattgacga tctattattg ccaggaaaca
     2041 cgaatttacc atggacttca ggatccttgg gaagaggtag aagatccaaa ttgtaataac
     2101 tcgagtggcg cgaatttctt atgatttatg atttttatta ttaaataagt tataaaaaaa
     2161 ataagtgtat acaaatttta aagtgactct taggttttaa aacgaaaatt cttattcttg
     2221 agtaactctt tcctgtaggt caggttgctt tctcaggtat agcatgaggt cgctcttatt
     2281 gaccacacct ctaccggcat gccgagcaaa tgcctgcaaa tcgctcccca tttcgctgcc
     2341 aaaccagatg tcaacacagc tacaacgttg ataggtcaag atcaatgtaa acaattactt
     2401 tgttatgtag agttttttta gctacctata ttccaccata acatcaatca tgcggttgct
     2461 ggtgtattta ccaataatgt ttaatgtata tatatatata tatatatggg gccgtatact
     2521 tacatatagt agatgtcaag cgtaggcgct tcccctgccg gctgtgaggg cgccataacc
     2581 aaggtatcta tagaccgcca atcagcaaac tacctccgta cattcatgtt gcacccacac
     2641 atttatacac ccagaccgcg acaaattacc cataaggttg tttgtgacgg cgtcgtacaa
     2701 gagaacgtgg gaacttttta ggctcaccaa aaaagaaaga aaaaatacga gttgctgaca
     2761 gaagcctcaa gaaaaaaaaa attcttcttc gactatgctg gaggcagaga tgatcgagcc
     2821 ggtagttaac tatatatagc taaattggtt ccatcacctt cttttctggt gtcgctcctt
     2881 ctagtgctat ttctggcttt tcctattttt ttttttccat ttttctttct ctctttctaa
     2941 tatataaatt ctcttgcatt ttctattttt ctctctatct attctacttg tttattccct
     3001 tcaaggtttt tttttaagga gtacttgttt ttagaatata cggtcaacga actataatta
     3061 actaaacaga tctatgtcag agttgagagc cttcagtgcc ccagggaaag cgttactagc
     3121 tggtggatat ttagttttag atacaaaata tgaagcattt gtagtcggat tatcggcaag
     3181 aatgcatgct gtagcccatc cttacggttc attgcaaggg tctgataagt ttgaagtgcg
     3241 tgtgaaaagt aaacaattta aagatgggga gtggctgtac catataagtc ctaaaagtgg
     3301 cttcattcct gtttcgatag gcggatctaa gaaccctttc attgaaaaag ttatcgctaa
     3361 cgtatttagc tactttaaac ctaacatgga cgactactgc aatagaaact tgttcgttat
     3421 tgatattttc tctgatgatg cctaccattc tcaggaggat agcgttaccg aacatcgtgg
     3481 caacagaaga ttgagttttc attcgcacag aattgaagaa gttcccaaaa cagggctggg
     3541 ctcctcggca ggtttagtca cagttttaac tacagctttg gcctcctttt ttgtatcgga
     3601 cctggaaaat aatgtagaca aatatagaga agttattcat aatttagcac aagttgctca
     3661 ttgtcaagct cagggtaaaa ttggaagcgg gtttgatgta gcggcggcag catatggatc
     3721 tatcagatat agaagattcc cacccgcatt aatctctaat ttgccagata ttggaagtgc
     3781 tacttacggc agtaaactgg cgcatttggt tgatgaagaa gactggaata ttacgattaa
     3841 aagtaaccat ttaccttcgg gattaacttt atggatgggc gatattaaga atggttcaga
     3901 aacagtaaaa ctggtccaga aggtaaaaaa ttggtatgat tcgcatatgc cagaaagctt
     3961 gaaaatatat acagaactcg atcatgcaaa ttctagattt atggatggac tatctaaact
     4021 agatcgctta cacgagactc atgacgatta cagcgatcag atatttgagt ctcttgagag
     4081 gaatgactgt acctgtcaaa agtatcctga aatcacagaa gttagagatg cagttgccac
     4141 aattagacgt tcctttagaa aaataactaa agaatctggt gccgatatcg aacctcccgt
     4201 acaaactagc ttattggatg attgccagac cttaaaagga gttcttactt gcttaatacc
     4261 tggtgctggt ggttatgacg ccattgcagt gattactaag caagatgttg atcttagggc
     4321 tcaaaccgct aatgacaaaa gattttctaa ggttcaatgg ctggatgtaa ctcaggctga
     4381 ctggggtgtt aggaaagaaa aagatccgga aacttatctt gataaaggat ccttgggaag
     4441 aggtagaaga tccaaattgt aataactcga gtggcagtgc ttttaactaa gaattattag
     4501 tcttttctgc ttattttttc atcatagttt agaacacttt atattaacga atagtttatg
     4561 aatctattta ggtttaaaaa ttgatacagt tttataagtt actttttcaa agactcgtgc
     4621 tgtctattgc ataatgcact ggaaggggaa aaaaaaggtg cacacgcgtg gctttttctt
     4681 gaatttgcag tttgaaaaat gctggatgca cacactggct taagatgaca acggtgagta
     4741 aggaaagagt gaggaactat cgcatacctg catttaaaga tgccgatttg ggcgcgaatc
     4801 ctttattttg gcttcaccct catactatta tcagggccag aaaaaggaag tgtttccctc
     4861 cttcttgaat tgatgttacc ctcataaagc acgtggcctc ttatcgagaa agaaattacc
     4921 gtcgctcgtg atttgtttgc aaaaagaaca aaactgaaaa aacccagaca cgctcgactt
     4981 cctgtcatcc tattgattgc agcttccaat ttcgtcacac aacaaggtcc tagcgacggc
     5041 tcacaggttt tgtaacaagc aatcgaaggt tctggaatgg cgggaaaggg tttagtacca
     5101 catgctatga tgcccactgt gatctccaga gcaaagttcg ttcgatcgta ctgttactct
     5161 ctctctttca aacagaattg tccgaatcgt gtgacaacaa cagcctgttc tcacacactc
     5221 ttttcttcta accaaggggg tggtttagtt tagtagaacc tcgtgaaact tacatttaca
     5281 tatatataaa cttgcataaa ttggtcaatg caagaaatac atatttggtc ttttctaatt
     5341 cgtagttttt caagttctta gatgctttct ttttctcttt tttacagatc atcaaggaag
     5401 taattatcta ctttttacaa caaatataaa acaagatcta tgaccgttta cacagcatcc
     5461 gttaccgcac ccgtcaacat cgcaaccctt aagtattggg ggaaaaggga cacgaagttg
     5521 aatctgccca ccaattcgtc catatcagtg actttatcgc aagatgacct cagaacgttg
     5581 acctctgcgg ctactgcacc tgagtttgaa cgcgacactt tgtggttaaa tggagaacca
     5641 cacagcatcg acaatgaaag aactcaaaat tgtctgcgcg acctacgcca attaagaaag
     5701 gaaatggaat cgaaggacgc ctcattgccc acattatctc aatggaaact ccacattgtc
     5761 tccgaaaata actttcctac agcagctggt ttagcttcct ccgctgctgg ctttgctgca
     5821 ttggtttctg caattgctaa gttataccaa ttaccacagt caacttcaga aatatctaga
     5881 atagcaagaa aggggtctgg ttcagcttgt agatcgttgt ttggcggata cgtggcctgg
     5941 gaaatgggaa aagctgaaga tggtcatgat tccatggcag tacaaatcgc agacagctct
     6001 gactggcctc agatgaaagc ttgtgtccta gttgtcagcg atattaaaaa ggatgtgagt
     6061 tccactcagg gtatgcaatt gaccgtggca acctccgaac tatttaaaga aagaattgaa
     6121 catgtcgtac caaagagatt tgaagtcatg cgtaaagcca ttgttgaaaa agatttcgcc
     6181 acctttgcaa aggaaacaat gatggattcc aactctttcc atgccacatg tttggactct
     6241 ttccctccaa tattctacat gaatgacact tccaagcgta tcatcagttg gtgccacacc
     6301 attaatcagt tttacggaga aacaatcgtt gcatacacgt ttgatgcagg tccaaatgct
     6361 gtgttgtact acttagctga aaatgagtcg aaactctttg catttatcta taaattgttt
     6421 ggctctgttc ctggatggga caagaaattt actactgagc agcttgaggc tttcaaccat
     6481 caatttgaat catctaactt tactgcacgt gaattggatc ttgagttgca aaaggatgtt
     6541 gccagagtga ttttaactca agtcggttca ggcccacaag aaacaaacga atctttgatt
     6601 gacgcaaaga ctggtctacc aaaggaagga tccttgggaa gaggtagaag atccaaattg
     6661 taataactcg agtggcataa agcaatcttg atgaggataa tgattttttt ttgaatatac
     6721 ataaatacta ccgtttttct gctagatttt gtgatgacgt aaataagtac atattacttt
     6781 ttaagccaag acaagattaa gcattaactt tacccttttc tttctaagtt tcaatattag
     6841 ttatcactgt ttaaaagtta tggcgagaac gtcggcggtt aaaatatatt accctgaacg
     6901 gctgagcagt tacagagatg ttacgaacca ctagtgcact gcagtacaat ctagctagaa
     6961 gttttgtagg tatatgtgat ttaagatata gataataaaa gatatatata tgtaaatatt
     7021 atgaagggga tattagcgta atttaaacta ttgaaatacc ggcttcaata tttcagctaa
     7081 cttgccattt ttcttcaaag tttccaaatc gctgttacca ccaatgtgct tgccattgat
     7141 gtatacgtta ggtacagttt tttggcccga gatttcttct aaagcgtctt gaatctctga
     7201 gccattgctc atttcatcta attccaacac aagggccttg gatttgggaa cgttcaattc
     7261 ttggaagagg gtagacaaag tagctttaca gtaagggcag tatgtctttg ctgcaacaaa
     7321 cacttccttt tggccaatca gatcctttac gtgagcaact gtttcctggg ataccatttt
     7381 tggagtagat aaaaatcttt tagcaataat tcttgtggca aacaacgtga taatgataat
     7441 aacaattacc gagcggccgc gtgttacaac caattaacca attctgatta gaaaaactca
     7501 tcgagcatca aatgaaactg caatttattc atatcaggat tatcaatacc atatttttga
     7561 aaaagccgtt tctgtaatga aggagaaaac tcaccgaggc agttccatag gatggcaaga
     7621 tcctggtatc ggtctgcgat tccgactcgt ccaacatcaa tacaacctat taatttcccc
     7681 tcgtcaaaaa taaggttatc aagtgagaaa tcaccatgag tgacgactga atccggtgag
     7741 aatggcaaaa gcttatgcat ttctttccag acttgttcaa caggccagcc attacgctcg
     7801 tcatcaaaat cactcgcatc aaccaaaccg ttattcattc gtgattgcgc ctgagcgagg
     7861 cgaaatacgc gatcgctgtt aaaaggacaa ttacaaacag gaatcgaatg caaccggcgc
     7921 aggaacactg ccagcgcatc aacaatattt tcacctgaat caggatattc ttctaatacc
     7981 tggaatgctg ttttcccggg gatcgcagtg gtgagtaacc atgcatcatc aggagtacgg
     8041 ataaaatgct tgatggtcgg aagaggcata aattccgtca gccagtttag tctgaccatc
     8101 tcatctgtaa catcattggc aacgctacct ttgccatgtt tcagaaacaa ctctggcgca
     8161 tcgggcttcc catacaatcg atagattgtc gcacctgatt gcccgacatt atcgcgagcc
     8221 catttatacc catataaatc agcatccatg ttggaattta atcgcggcct ggagcaagac
     8281 gtttcccgtt gaatatggct cataacaccc cttgtattac tgtttatgta agcagacagt
     8341 tttattgttc atgatgatat atttttatct tgtgcaatgt aacatcagag attttgagac
     8401 acaacgtggc tttgttgaat aaatcgaact tttgctgagt tgaaggatca gtcatgacca
     8461 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     8521 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     8581 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     8641 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     8701 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     8761 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     8821 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     8881 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     8941 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     9001 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     9061 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     9121 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     9181 ttcctgcgtt atcccctgat tctgtggata accgtgcggc cgccaatgtt tgtttcttct
     9241 tatcttcagc tgctgaggat gtattttttg gattatcatt gaggttttgc gatgacattt
     9301 ttgaattcac tttatgattg attttcttta tcttggttat atttttgggc tctctcgttt
     9361 atttaacgcc accctaaatt gtgtcttaac aatacgcttc ctctcagtat ttttttttaa
     9421 ataaaaaaat atgatagaac tgctgaacac tactttttaa aaccttttaa taaatagaca
     9481 agtgtcaacc aaggatgtgc caattttcct ttgtgattca gctgcgcttg tatgtgcttc
     9541 cagctttact tttactaatt ttttcataag cacaaaatag taagatcttt tttctcttgg
     9601 cggtgaaaaa ttttgattcg atggatcaat aacatactgc aaaaaacggt ttgaggacat
     9661 atgaatactg catttactag taatttgcct agtatatagt ttgactacac agattataag
     9721 ctattgtccc tgaattcgca tctagactga
//
