LOCUS       sequence-475876-        6764 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express Idi1p and Erg20WW for synthesis of GPP in the peroxisome; 
            chromosomal integration by CRISPR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6764)
  TITLE     Dueber Lab plasmids 2025
REFERENCE   2  (bases 1 to 6764)
  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: pKS1466
FEATURES             Location/Qualifiers
     source          1..6764
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(4502..5317)
                     /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(4502..5317)
                     /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     5225..5244
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5309..5328
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5542..6130
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6031..6050
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 actggccgat aattgcagac gaacgtgtgg agtgtttgct tggattcttt agtaaaaggg
       61 gaagaacagt tggaagggcc aaagtggaag tcacaaaaca gtggtcctat ataaaagaac
      121 aagaaaaaga ttatttatat acaactgcgg tcacaagaag caacgcgaga gagcacaaca
      181 cgctgttatc acgcaaacta tgttttgaca ccgagccata gccgtgattg tgcgtcacat
      241 tgggcgataa tgaacgctaa atgaccaact cccatccgta ggagcccctt agggcgtgcc
      301 aatagtttca cgcgcttaat gcgaagtgct cggaacggac aactgtggtc gtttggcacc
      361 gggaaagtgg tactagaccg agagtttcgc atttgtatgg caggacgttc tgggagcttc
      421 gcgtctaaag ctttttcggg cgcgaaatgc agaccagacc agaacaaaac aactgacaag
      481 aaggcgttta atttaatatg ttgttcactc gcgcctgggc tgttgttatt cggctagata
      541 catacgtgtt tgtgcgtatg tagttatatc atatataagt atattaggat gaggcggtga
      601 aagagatttt ttttttttcg cttaatttat tcttttctct atcttttttc ctacatcttg
      661 ttcaaaagag tagcaaaaac aacaatcaat acaataaaat aagatctatg actgccgaca
      721 acaatagtat gccccatggt gcagtatcta gttacgccaa attagtgcaa aaccaaacac
      781 ctgaagacat tttggaagag tttcctgaaa ttattccatt acaacaaaga cctaataccc
      841 gatctagtga aacgtcaaat gacgaaagcg gagaaacatg tttttctggt catgatgagg
      901 agcaaattaa gttaatgaat gaaaattgta ttgttttgga ttgggacgat aatgctattg
      961 gtgccggtac caagaaagtt tgtcatttaa tggaaaatat tgaaaagggt ttactacatc
     1021 gtgcattctc cgtctttatt ttcaatgaac aaggtgaatt acttttacaa caaagagcca
     1081 ctgaaaaaat aactttccct gatctttgga ctaacacatg ctgctctcat ccactatgta
     1141 ttgatgacga attaggtttg aagggtaagc tagacgataa gattaagggc gctattactg
     1201 cggcggtgag aaaactagat catgaattag gtattccaga agatgaaact aagacaaggg
     1261 gtaagtttca ctttttaaac agaatccatt acatggcacc aagcaatgaa ccatggggtg
     1321 aacatgaaat tgattacatc ctattttata agatcaacgc taaagaaaac ttgactgtca
     1381 acccaaacgt caatgaagtt agagacttca aatgggtttc accaaatgat ttgaaaacta
     1441 tgtttgctga cccaagttac aagtttacgc cttggtttaa gattatttgc gagaattact
     1501 tattcaactg gtgggagcaa ttagatgacc tttctgaagt ggaaaatgac aggcaaattc
     1561 atagaatgct aggatccttg ggaagaggta gaagatccaa attgtaataa ctcgagtggc
     1621 attgaattga attgaaatcg atagatcaat ttttttcttt tctctttccc catcctttac
     1681 gctaaaataa tagtttattt tattttttga atatttttta tttatatacg tatatataga
     1741 ctattattta tcttttaatg attattaaga tttttattaa aaaaaaattc gctcctcttt
     1801 taatgccttt atgcagtttt tttttcccat tcgatatttc tatgtgctgc caaaccagat
     1861 gtcaacacag ctacaacgca cccatgaacc acacggttag tccaaaaggg gcagttcaga
     1921 ttccagatgc gggaattagc ttgctgccac cctcacctca ctaacgctgc ggtgtgcgga
     1981 tacttcatgc tatttataga cgcgcgtgtc ggaatcagca cgcgcaagaa ccaaatggga
     2041 aaatcggaat gggtccagaa ctgctttgag tgctggctat tggcgtctga tttccgtttt
     2101 gggaatcctt tgccgcgcgc ccctctcaaa actccgcaca agtcccagaa agcgggaaag
     2161 aaataaaacg ccaccaaaaa aaaaaaaata aaagccaatc ctcgaagcgt gggtggtagg
     2221 ccctggatta tcccgtacaa gtatttctca ggagtaaaaa aaccgtttgt tttggaattt
     2281 cccatttcgc ggccacctac gccgctatct ttgcaacaac tatctgcgat aactcagcaa
     2341 attttgcata ttcgtgttgc agtattgcga taatgggagt cttacttcca acataacggc
     2401 agaaagaaat gtgagaaaat tttgcatcct ttgcctccgt tcaagtatat aaagtcggca
     2461 tgcttgataa tctttctttc catcctacat tgttctaatt attcttattc tcctttattc
     2521 tttcctaaca taccaagaaa ttaatcttct gtcattcgct taaacactat atcaataaag
     2581 atctatggct tcagaaaaag aaattaggag agagagattc ttgaacgttt tccctaaatt
     2641 agtagaggaa ttgaacgcat cgcttttggc ttacggtatg cctaaggaag catgtgactg
     2701 gtatgcccac tcattgaact acaacactcc aggcggtaag ctaaatagag gtttgtccgt
     2761 tgtggacacg tatgctattc tctccaacaa gaccgttgaa caattggggc aagaagaata
     2821 cgaaaaggtt gccattctag gttggtgcat tgagttgttg caggcttact ggttggtcgc
     2881 cgatgatatg atggacaagt ccattaccag aagaggccaa ccatgttggt acaaggttcc
     2941 tgaagttggg gaaattgcca tctgggacgc attcatgtta gaggctgcta tctacaagct
     3001 tttgaaatct cacttcagaa acgaaaaata ctacatagat atcaccgaat tgttccatga
     3061 ggtcaccttc caaaccgaat tgggccaatt gatggactta atcactgcac ctgaagacaa
     3121 agtcgacttg agtaagttct ccctaaagaa gcactccttc atagttactt tcaagactgc
     3181 ttactattct ttctacttgc ctgtcgcatt ggccatgtac gttgccggta tcacggatga
     3241 aaaggatttg aaacaagcca gagatgtctt gattccattg ggtgaatact tccaaattca
     3301 agatgactac ttagactgct tcggtacccc agaacagatc ggtaagatcg gtacagatat
     3361 ccaagataac aaatgttctt gggtaatcaa caaggcattg gaacttgctt ccgcagaaca
     3421 aagaaagact ttagacgaaa attacggtaa gaaggactca gtcgcagaag ccaaatgcaa
     3481 aaagattttc aatgacttga aaattgaaca gctataccac gaatatgaag agtctattgc
     3541 caaggatttg aaggccaaaa tttctcaggt cgatgagtct cgtggcttca aagctgatgt
     3601 cttaactgcg ttcttgaaca aagtttacaa gagaagcaaa ggatccttgg gaagaggtag
     3661 aagatccaaa ttgtaataac tcgagtggcg ccaattggtg cggcaattga taataacgaa
     3721 aatgtctttt aatgatctgg gtataatgag gaattttccg aacgttttta ctttatatat
     3781 atatatacat gtaacatata ttctatacgc tatagagaaa ggaaattttt caattaaaaa
     3841 aaaaatagag aaagagtttc acttcttgat tatcgctaac actaatggtt gaagtactgc
     3901 tactttaatt ttatgctgag cagttacaga gatgttacga accactagtg cactgcagta
     3961 catgacatgt atgggttgaa aatatttaga ggatgctaaa agtcccgtct aaaatagagg
     4021 aagtcatgga ttacattatc acctttattg tcctgttccg tctggctaat ttgggacgag
     4081 gaacgtttgt ctccattcgc aaccttttga aaccatatga acggcggcta atctgacaga
     4141 tgtcttatat ttctaggatt gtagaaagcc tcctcatcaa accaccttgt ttgaaaaagg
     4201 cactagtgcg gttgctttac ttgatctagt attctcctac cttggagcgg tcccatatca
     4261 actaaattca gttttctgac tgagttaaaa actaatgtag cgacaccact tccattggtg
     4321 cctccgttat tttttttttg tatgttaatg ctaaataata caccgctatg tatttcaggg
     4381 cactacttct acacatcaac ggtactaaac atttcgctca agatcggtcc gctttcacgt
     4441 atttggatgc tatgcaatgt tgccgagcgg ccgcgtgtta caaccaatta accaattctg
     4501 attagaaaaa ctcatcgagc atcaaatgaa actgcaattt attcatatca ggattatcaa
     4561 taccatattt ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg aggcagttcc
     4621 ataggatggc aagatcctgg tatcggtctg cgattccgac tcgtccaaca tcaatacaac
     4681 ctattaattt cccctcgtca aaaataaggt tatcaagtga gaaatcacca tgagtgacga
     4741 ctgaatccgg tgagaatggc aaaagcttat gcatttcttt ccagacttgt tcaacaggcc
     4801 agccattacg ctcgtcatca aaatcactcg catcaaccaa accgttattc attcgtgatt
     4861 gcgcctgagc gaggcgaaat acgcgatcgc tgttaaaagg acaattacaa acaggaatcg
     4921 aatgcaaccg gcgcaggaac actgccagcg catcaacaat attttcacct gaatcaggat
     4981 attcttctaa tacctggaat gctgttttcc cggggatcgc agtggtgagt aaccatgcat
     5041 catcaggagt acggataaaa tgcttgatgg tcggaagagg cataaattcc gtcagccagt
     5101 ttagtctgac catctcatct gtaacatcat tggcaacgct acctttgcca tgtttcagaa
     5161 acaactctgg cgcatcgggc ttcccataca atcgatagat tgtcgcacct gattgcccga
     5221 cattatcgcg agcccattta tacccatata aatcagcatc catgttggaa tttaatcgcg
     5281 gcctggagca agacgtttcc cgttgaatat ggctcataac accccttgta ttactgttta
     5341 tgtaagcaga cagttttatt gttcatgatg atatattttt atcttgtgca atgtaacatc
     5401 agagattttg agacacaacg tggctttgtt gaataaatcg aacttttgct gagttgaagg
     5461 atcagtcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     5521 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     5581 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     5641 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     5701 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     5761 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     5821 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     5881 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     5941 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     6001 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     6061 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     6121 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     6181 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgtg cggccgccaa
     6241 ttgaaaaagg ccactactga gccaagcgca gaaccagatg agccattcat tggatacttg
     6301 ggtgacgtta ctgcatctat aaaccaagat ataaaagaat atggtcgcag aagaacagta
     6361 atacctacaa gcgatttagt agcgtcgttc tacaccaaaa agacgcctgt ttttaaggtc
     6421 aagttcagca gaagcaattt agcccttgca ggaggtgctt tcaggccata attcacactg
     6481 aaaaaaataa cagcatggaa gtgatatgta ataagtataa cctaatccga tatgtaacta
     6541 ttagatgctt tttcgacatc gtacttgacc tgctcggcgg ttatcatagc gtcttcttta
     6601 gccttttttt tttcgctcag tcatcggggc caaaacaaga aaggcgaaga acagaaaaat
     6661 gaaaaatgta actggttgtt gtaaaaacct tccagagaaa taagatgaaa tgtatgttgc
     6721 tggcacgaac gaagacaaat accctgaatt cgcatctaga ctga
//
