LOCUS       sequence-475886-        7554 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express CpPT1 membrane protein cargo fused to PTS1 and 
            Pex22(1-36)-mRuby2; chromosomal integration by CRISPR .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7554)
  TITLE     Dueber Lab plasmids 2025
REFERENCE   2  (bases 1 to 7554)
  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: pKS1613
FEATURES             Location/Qualifiers
     source          1..7554
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        3158..3571
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             3705..4412
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby, with improved photophysical properties (Lam et al., 
                     2012)"
                     /label=mRuby2
                     /translation="VSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGNPYMGTQTMR
                     IKVIEGGPLPFAFDILATSFMYGSRTFIKYPKGIPDFFKQSFPEGFTWERVTRYEDGGV
                     VTVMQDTSLEDGCLVYHVQVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTHM
                     ALKVDGGGHLSCSFVTTYRSKKTVGNIKMPGIHAVDHRLERLEESDNEMFVVQREHAVA
                     KFAGLGGGMDELYK"
     CDS             complement(5292..6107)
                     /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(5292..6107)
                     /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     6015..6034
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6099..6118
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      6332..6920
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6821..6840
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 actggccgat aattgcagac gaacgactag agatctatgt gaggatccta actcgagatc
       61 gagctgccaa accagatgtc aacacagcta caacgttgat aggtcaagat caatgtaaac
      121 aattactttg ttatgtagag tttttttagc tacctatatt ccaccataac atcaatcatg
      181 cggttgctgg tgtatttacc aataatgttt aatgtatata tatatatata tatatggggc
      241 cgtatactta catatagtag atgtcaagcg taggcgcttc ccctgccggc tgtgagggcg
      301 ccataaccaa ggtatctata gaccgccaat cagcaaacta cctccgtaca ttcatgttgc
      361 acccacacat ttatacaccc agaccgcgac aaattaccca taaggttgtt tgtgacggcg
      421 tcgtacaaga gaacgtggga actttttagg ctcaccaaaa aagaaagaaa aaatacgagt
      481 tgctgacaga agcctcaaga aaaaaaaaat tcttcttcga ctatgctgga ggcagagatg
      541 atcgagccgg tagttaacta tatatagcta aattggttcc atcaccttct tttctggtgt
      601 cgctccttct agtgctattt ctggcttttc ctattttttt ttttccattt ttctttctct
      661 ctttctaata tataaattct cttgcatttt ctatttttct ctctatctat tctacttgtt
      721 tattcccttc aaggtttttt tttaaggagt acttgttttt agaatatacg gtcaacgaac
      781 tataattaac taaacagatc tatggtttct aaaggtgaag aattattcac tggtgttgtc
      841 ccaattttgg ttgaattaga tggtgatgtt aatggtcaca aattttctgt ctccggtgaa
      901 ggtgaaggtg atgctactta cggtaaattg accttaaaat ttatttgtac tactggtaaa
      961 ttgccagttc catggccaac cttagtcact actttatctt ggggtgttca atgttttgca
     1021 agatacccag atcatatgaa acaacatgac tttttcaagt ctgccatgcc agaaggttat
     1081 gttcaagaaa gaactatttt tttcaaagat gacggtaact acaagaccag agctgaagtc
     1141 aagtttgaag gtgatacctt agttaataga atcgaattaa aaggtattga ttttaaagaa
     1201 gatggtaaca ttttaggtca caaattggaa tacaattatt tctctgacaa tgtttacatc
     1261 actgctgaca aacaaaagaa tggtatcaaa gctaacttca aaattagaca caacattgaa
     1321 gatggtggtg ttcaattagc tgaccattat caacaaaata ctccaattgg tgatggtcca
     1381 gtcttgttac cagacaacca ttacttatcc actcaatcta agttatccaa agatccaaac
     1441 gaaaagaggg accacatggt cttgttagaa tttgttactg ctgctggtat taccttgggt
     1501 atggatgaat tgtacaaagg ttctattcag tcttcattct gtttaacaaa taataagata
     1561 ggtaacaatg aagatatgat gaatagatac cataaaccat taaagaaatc tacagttcct
     1621 atggctttac aagatgactc tgctacaaaa tctcaaaacg aaaatattgt ttctacatct
     1681 ttcttggact tcttaaccaa gaagttggat gcattttata ggttatcaag accatatgct
     1741 tggacttcaa tcattgtagg tattttgtca tcttctttgt tgccaatcca atctttggca
     1801 gatttgactc ctaccttctt gatcgaggta ttgaagccta tagttccttc aattatgatg
     1861 aatatatttg tcgttgctat taatcagtta tcagatatag caattgataa ggttaacaag
     1921 ccaaacttgc ctttagtttc tggtgacata tctattggtg aggctatagc tattgctgtt
     1981 atctcaacat tgacttcttt ggctatgggt gttatgttaa gatcaccacc attagttata
     2041 gcattaatat tgaggtgtat cttaggtgca gcttattcta ttgatttgcc attattgaga
     2101 tggaaggctt ctcctttgat ggctgccgtc gcaatcgtta ttggaaatgg tataaataat
     2161 gttttgccat acttcttaca cgtccaaaag tacgttttag gtaggcctgt tgtctttact
     2221 aagcctttat tattcgcagt tgcattctca gcaatatttt ctatcgtctt atcattctta
     2281 aaagacattc ctgacgttga aggagataag aaatctggta tcagaacatt accagtcatt
     2341 ttgggtaaag agagagtctt gtctatgtca actggtattt tgttgatggc ctatgcttct
     2401 gccgccttgg ctggagtctt ctcaccaatt ttattatgta aattggtaac tatgattggt
     2461 cattcagtat tgggttttat cttatggtct aaggcccaaa cagttgattt atcaaatgct
     2521 aaatctacat attcattctt cttattcatt ttccagttgt attacaccga gttcttctta
     2581 atgcattttg taagaggatc ctccaaattg taataactcg agtggcagtg cttttaacta
     2641 agaattatta gtcttttctg cttatttttt catcatagtt tagaacactt tatattaacg
     2701 aatagtttat gaatctattt aggtttaaaa attgatacag ttttataagt tactttttca
     2761 aagactcgtg ctgtctattg cataatgcac tggaagggga aaaaaaaggt gcacacgcgt
     2821 ggctttttct tgaatttgca gtttgaaaaa tgctggatgc acacactggc ttaagatgac
     2881 aacgccttgc caacagggag ttcttcagag acatggaggc tcaaaacgaa attattgaca
     2941 gcctagacat caatagtcat acaacagaaa gcgaccaccc aactttggct gataatagcg
     3001 tataaacaat gcatactttg tacgttcaaa atacaatgca gtagatatat ttatgcatat
     3061 tacatataat acatatcaca taggaagcaa caggcgcgtt ggacttttaa ttttcgagga
     3121 ccgcgaatcc ttacatcaca cccaatcccc cacaagtgat cccccacaca ccatagcttc
     3181 aaaatgtttc tactcctttt ttactcttcc agattttctc ggactccgcg catcgccgta
     3241 ccacttcaaa acacccaagc acagcatact aaatttcccc tctttcttcc tctagggtgt
     3301 cgttaattac ccgtactaaa ggtttggaaa agaaaaaaga caccgcctcg tttctttttc
     3361 ttcgtcgaaa aaggcaataa aaatttttat cacgtttctt tttcttgaaa attttttttt
     3421 ttgatttttt tctctttcga tgacctccca ttgatattta agttaataaa cggtcatcaa
     3481 tttctcaagt ttcagtttca tttttcttgt tctattacaa ctttttttac ttcttgctca
     3541 ttagaaagaa agcatagcaa tctaatctaa gttttaatta caaaagatct atgccaccac
     3601 catcaagaag tagaataaac aaaacaagaa cattaggaat agtgggtaca gctatagcag
     3661 tgttggtcac gtcctactat atatatcaaa aggtgacagg ttctgtgtcc aaaggagagg
     3721 agttaatcaa ggaaaacatg agaatgaaag ttgtcatgga gggctccgtt aatggtcacc
     3781 aattcaagtg tacaggggaa ggtgaaggta atccttacat gggtacacaa actatgagaa
     3841 ttaaagtaat tgaaggcgga ccactaccat ttgcatttga cattctggca acgtcattca
     3901 tgtacggatc acgaactttc atcaagtacc ctaaaggtat accagacttt ttcaagcaat
     3961 cttttccaga gggttttaca tgggaaaggg ttacaagata cgaagatggg ggtgtcgtca
     4021 cagttatgca agatacttca ttagaagatg gctgccttgt ctatcatgtg caagtaagag
     4081 gggtgaattt tccttctaac ggacctgtga tgcagaaaaa gaccaaaggt tgggaaccaa
     4141 atactgaaat gatgtaccca gctgatggag gtttgagagg ctacacacac atggcgctta
     4201 aagttgatgg tggaggtcat ttgtcttgta gttttgttac cacttatcgt tctaaaaaga
     4261 ctgttggcaa tatcaaaatg ccaggaatac atgctgtaga ccacagacta gaaagactcg
     4321 aagagagcga taacgaaatg ttcgttgtac agagagagca tgccgtagcc aaatttgctg
     4381 gcttaggcgg tggtatggat gaattgtata agggatccgg gtcaggctcc ggatctggcc
     4441 ctcccccagc tcttccgcca aaaagacgaa ggtaatggca gtgcttttaa ctaagaatta
     4501 ttagtctttt ctgcttattt tttcatcata gtttagaaca ctttatatta acgaatagtt
     4561 tatgaatcta tttaggttta aaaattgata cagttttata agttactttt tcaaagactc
     4621 gtgctgtcta ttgcataatg cactggaagg ggaaaaaaaa ggtgcacacg cgtggctttt
     4681 tcttgaattt gcagtttgaa aaatgctgag cagttacaga gatgttacga accactagtg
     4741 cactgcagta catgacatgt atgggttgaa aatatttaga ggatgctaaa agtcccgtct
     4801 aaaatagagg aagtcatgga ttacattatc acctttattg tcctgttccg tctggctaat
     4861 ttgggacgag gaacgtttgt ctccattcgc aaccttttga aaccatatga acggcggcta
     4921 atctgacaga tgtcttatat ttctaggatt gtagaaagcc tcctcatcaa accaccttgt
     4981 ttgaaaaagg cactagtgcg gttgctttac ttgatctagt attctcctac cttggagcgg
     5041 tcccatatca actaaattca gttttctgac tgagttaaaa actaatgtag cgacaccact
     5101 tccattggtg cctccgttat tttttttttg tatgttaatg ctaaataata caccgctatg
     5161 tatttcaggg cactacttct acacatcaac ggtactaaac atttcgctca agatcggtcc
     5221 gctttcacgt atttggatgc tatgcaatgt tgccgagcgg ccgcgtgtta caaccaatta
     5281 accaattctg attagaaaaa ctcatcgagc atcaaatgaa actgcaattt attcatatca
     5341 ggattatcaa taccatattt ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg
     5401 aggcagttcc ataggatggc aagatcctgg tatcggtctg cgattccgac tcgtccaaca
     5461 tcaatacaac ctattaattt cccctcgtca aaaataaggt tatcaagtga gaaatcacca
     5521 tgagtgacga ctgaatccgg tgagaatggc aaaagcttat gcatttcttt ccagacttgt
     5581 tcaacaggcc agccattacg ctcgtcatca aaatcactcg catcaaccaa accgttattc
     5641 attcgtgatt gcgcctgagc gaggcgaaat acgcgatcgc tgttaaaagg acaattacaa
     5701 acaggaatcg aatgcaaccg gcgcaggaac actgccagcg catcaacaat attttcacct
     5761 gaatcaggat attcttctaa tacctggaat gctgttttcc cggggatcgc agtggtgagt
     5821 aaccatgcat catcaggagt acggataaaa tgcttgatgg tcggaagagg cataaattcc
     5881 gtcagccagt ttagtctgac catctcatct gtaacatcat tggcaacgct acctttgcca
     5941 tgtttcagaa acaactctgg cgcatcgggc ttcccataca atcgatagat tgtcgcacct
     6001 gattgcccga cattatcgcg agcccattta tacccatata aatcagcatc catgttggaa
     6061 tttaatcgcg gcctggagca agacgtttcc cgttgaatat ggctcataac accccttgta
     6121 ttactgttta tgtaagcaga cagttttatt gttcatgatg atatattttt atcttgtgca
     6181 atgtaacatc agagattttg agacacaacg tggctttgtt gaataaatcg aacttttgct
     6241 gagttgaagg atcagtcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     6301 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     6361 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     6421 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     6481 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     6541 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     6601 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     6661 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     6721 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     6781 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     6841 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     6901 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     6961 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgtg
     7021 cggccgccaa ttgaaaaagg ccactactga gccaagcgca gaaccagatg agccattcat
     7081 tggatacttg ggtgacgtta ctgcatctat aaaccaagat ataaaagaat atggtcgcag
     7141 aagaacagta atacctacaa gcgatttagt agcgtcgttc tacaccaaaa agacgcctgt
     7201 ttttaaggtc aagttcagca gaagcaattt agcccttgca ggaggtgctt tcaggccata
     7261 attcacactg aaaaaaataa cagcatggaa gtgatatgta ataagtataa cctaatccga
     7321 tatgtaacta ttagatgctt tttcgacatc gtacttgacc tgctcggcgg ttatcatagc
     7381 gtcttcttta gccttttttt tttcgctcag tcatcggggc caaaacaaga aaggcgaaga
     7441 acagaaaaat gaaaaatgta actggttgtt gtaaaaacct tccagagaaa taagatgaaa
     7501 tgtatgttgc tggcacgaac gaagacaaat accctgaatt cgcatctaga ctga
//
