LOCUS       sequence-475878-        6834 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express CpPT1 membrane protein cargo without mTurqoise2 marker 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 6834)
  TITLE     Dueber Lab plasmids 2025
REFERENCE   2  (bases 1 to 6834)
  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: pKS1612
FEATURES             Location/Qualifiers
     source          1..6834
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        2438..2851
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             2985..3692
                     /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(4572..5387)
                     /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(4572..5387)
                     /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     5295..5314
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5379..5398
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5612..6200
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6101..6120
                     /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 tatgattcag tcttcattct gtttaacaaa taataagata
      841 ggtaacaatg aagatatgat gaatagatac cataaaccat taaagaaatc tacagttcct
      901 atggctttac aagatgactc tgctacaaaa tctcaaaacg aaaatattgt ttctacatct
      961 ttcttggact tcttaaccaa gaagttggat gcattttata ggttatcaag accatatgct
     1021 tggacttcaa tcattgtagg tattttgtca tcttctttgt tgccaatcca atctttggca
     1081 gatttgactc ctaccttctt gatcgaggta ttgaagccta tagttccttc aattatgatg
     1141 aatatatttg tcgttgctat taatcagtta tcagatatag caattgataa ggttaacaag
     1201 ccaaacttgc ctttagtttc tggtgacata tctattggtg aggctatagc tattgctgtt
     1261 atctcaacat tgacttcttt ggctatgggt gttatgttaa gatcaccacc attagttata
     1321 gcattaatat tgaggtgtat cttaggtgca gcttattcta ttgatttgcc attattgaga
     1381 tggaaggctt ctcctttgat ggctgccgtc gcaatcgtta ttggaaatgg tataaataat
     1441 gttttgccat acttcttaca cgtccaaaag tacgttttag gtaggcctgt tgtctttact
     1501 aagcctttat tattcgcagt tgcattctca gcaatatttt ctatcgtctt atcattctta
     1561 aaagacattc ctgacgttga aggagataag aaatctggta tcagaacatt accagtcatt
     1621 ttgggtaaag agagagtctt gtctatgtca actggtattt tgttgatggc ctatgcttct
     1681 gccgccttgg ctggagtctt ctcaccaatt ttattatgta aattggtaac tatgattggt
     1741 cattcagtat tgggttttat cttatggtct aaggcccaaa cagttgattt atcaaatgct
     1801 aaatctacat attcattctt cttattcatt ttccagttgt attacaccga gttcttctta
     1861 atgcattttg taagaggatc ctccaaattg taataactcg agtggcagtg cttttaacta
     1921 agaattatta gtcttttctg cttatttttt catcatagtt tagaacactt tatattaacg
     1981 aatagtttat gaatctattt aggtttaaaa attgatacag ttttataagt tactttttca
     2041 aagactcgtg ctgtctattg cataatgcac tggaagggga aaaaaaaggt gcacacgcgt
     2101 ggctttttct tgaatttgca gtttgaaaaa tgctggatgc acacactggc ttaagatgac
     2161 aacgccttgc caacagggag ttcttcagag acatggaggc tcaaaacgaa attattgaca
     2221 gcctagacat caatagtcat acaacagaaa gcgaccaccc aactttggct gataatagcg
     2281 tataaacaat gcatactttg tacgttcaaa atacaatgca gtagatatat ttatgcatat
     2341 tacatataat acatatcaca taggaagcaa caggcgcgtt ggacttttaa ttttcgagga
     2401 ccgcgaatcc ttacatcaca cccaatcccc cacaagtgat cccccacaca ccatagcttc
     2461 aaaatgtttc tactcctttt ttactcttcc agattttctc ggactccgcg catcgccgta
     2521 ccacttcaaa acacccaagc acagcatact aaatttcccc tctttcttcc tctagggtgt
     2581 cgttaattac ccgtactaaa ggtttggaaa agaaaaaaga caccgcctcg tttctttttc
     2641 ttcgtcgaaa aaggcaataa aaatttttat cacgtttctt tttcttgaaa attttttttt
     2701 ttgatttttt tctctttcga tgacctccca ttgatattta agttaataaa cggtcatcaa
     2761 tttctcaagt ttcagtttca tttttcttgt tctattacaa ctttttttac ttcttgctca
     2821 ttagaaagaa agcatagcaa tctaatctaa gttttaatta caaaagatct atgccaccac
     2881 catcaagaag tagaataaac aaaacaagaa cattaggaat agtgggtaca gctatagcag
     2941 tgttggtcac gtcctactat atatatcaaa aggtgacagg ttctgtgtcc aaaggagagg
     3001 agttaatcaa ggaaaacatg agaatgaaag ttgtcatgga gggctccgtt aatggtcacc
     3061 aattcaagtg tacaggggaa ggtgaaggta atccttacat gggtacacaa actatgagaa
     3121 ttaaagtaat tgaaggcgga ccactaccat ttgcatttga cattctggca acgtcattca
     3181 tgtacggatc acgaactttc atcaagtacc ctaaaggtat accagacttt ttcaagcaat
     3241 cttttccaga gggttttaca tgggaaaggg ttacaagata cgaagatggg ggtgtcgtca
     3301 cagttatgca agatacttca ttagaagatg gctgccttgt ctatcatgtg caagtaagag
     3361 gggtgaattt tccttctaac ggacctgtga tgcagaaaaa gaccaaaggt tgggaaccaa
     3421 atactgaaat gatgtaccca gctgatggag gtttgagagg ctacacacac atggcgctta
     3481 aagttgatgg tggaggtcat ttgtcttgta gttttgttac cacttatcgt tctaaaaaga
     3541 ctgttggcaa tatcaaaatg ccaggaatac atgctgtaga ccacagacta gaaagactcg
     3601 aagagagcga taacgaaatg ttcgttgtac agagagagca tgccgtagcc aaatttgctg
     3661 gcttaggcgg tggtatggat gaattgtata agggatccgg gtcaggctcc ggatctggcc
     3721 ctcccccagc tcttccgcca aaaagacgaa ggtaatggca gtgcttttaa ctaagaatta
     3781 ttagtctttt ctgcttattt tttcatcata gtttagaaca ctttatatta acgaatagtt
     3841 tatgaatcta tttaggttta aaaattgata cagttttata agttactttt tcaaagactc
     3901 gtgctgtcta ttgcataatg cactggaagg ggaaaaaaaa ggtgcacacg cgtggctttt
     3961 tcttgaattt gcagtttgaa aaatgctgag cagttacaga gatgttacga accactagtg
     4021 cactgcagta catgacatgt atgggttgaa aatatttaga ggatgctaaa agtcccgtct
     4081 aaaatagagg aagtcatgga ttacattatc acctttattg tcctgttccg tctggctaat
     4141 ttgggacgag gaacgtttgt ctccattcgc aaccttttga aaccatatga acggcggcta
     4201 atctgacaga tgtcttatat ttctaggatt gtagaaagcc tcctcatcaa accaccttgt
     4261 ttgaaaaagg cactagtgcg gttgctttac ttgatctagt attctcctac cttggagcgg
     4321 tcccatatca actaaattca gttttctgac tgagttaaaa actaatgtag cgacaccact
     4381 tccattggtg cctccgttat tttttttttg tatgttaatg ctaaataata caccgctatg
     4441 tatttcaggg cactacttct acacatcaac ggtactaaac atttcgctca agatcggtcc
     4501 gctttcacgt atttggatgc tatgcaatgt tgccgagcgg ccgcgtgtta caaccaatta
     4561 accaattctg attagaaaaa ctcatcgagc atcaaatgaa actgcaattt attcatatca
     4621 ggattatcaa taccatattt ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg
     4681 aggcagttcc ataggatggc aagatcctgg tatcggtctg cgattccgac tcgtccaaca
     4741 tcaatacaac ctattaattt cccctcgtca aaaataaggt tatcaagtga gaaatcacca
     4801 tgagtgacga ctgaatccgg tgagaatggc aaaagcttat gcatttcttt ccagacttgt
     4861 tcaacaggcc agccattacg ctcgtcatca aaatcactcg catcaaccaa accgttattc
     4921 attcgtgatt gcgcctgagc gaggcgaaat acgcgatcgc tgttaaaagg acaattacaa
     4981 acaggaatcg aatgcaaccg gcgcaggaac actgccagcg catcaacaat attttcacct
     5041 gaatcaggat attcttctaa tacctggaat gctgttttcc cggggatcgc agtggtgagt
     5101 aaccatgcat catcaggagt acggataaaa tgcttgatgg tcggaagagg cataaattcc
     5161 gtcagccagt ttagtctgac catctcatct gtaacatcat tggcaacgct acctttgcca
     5221 tgtttcagaa acaactctgg cgcatcgggc ttcccataca atcgatagat tgtcgcacct
     5281 gattgcccga cattatcgcg agcccattta tacccatata aatcagcatc catgttggaa
     5341 tttaatcgcg gcctggagca agacgtttcc cgttgaatat ggctcataac accccttgta
     5401 ttactgttta tgtaagcaga cagttttatt gttcatgatg atatattttt atcttgtgca
     5461 atgtaacatc agagattttg agacacaacg tggctttgtt gaataaatcg aacttttgct
     5521 gagttgaagg atcagtcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     5581 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     5641 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     5701 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     5761 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     5821 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     5881 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     5941 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     6001 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     6061 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     6121 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     6181 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     6241 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgtg
     6301 cggccgccaa ttgaaaaagg ccactactga gccaagcgca gaaccagatg agccattcat
     6361 tggatacttg ggtgacgtta ctgcatctat aaaccaagat ataaaagaat atggtcgcag
     6421 aagaacagta atacctacaa gcgatttagt agcgtcgttc tacaccaaaa agacgcctgt
     6481 ttttaaggtc aagttcagca gaagcaattt agcccttgca ggaggtgctt tcaggccata
     6541 attcacactg aaaaaaataa cagcatggaa gtgatatgta ataagtataa cctaatccga
     6601 tatgtaacta ttagatgctt tttcgacatc gtacttgacc tgctcggcgg ttatcatagc
     6661 gtcttcttta gccttttttt tttcgctcag tcatcggggc caaaacaaga aaggcgaaga
     6721 acagaaaaat gaaaaatgta actggttgtt gtaaaaacct tccagagaaa taagatgaaa
     6781 tgtatgttgc tggcacgaac gaagacaaat accctgaatt cgcatctaga ctga
//
