LOCUS       sequence-360919-        6827 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vector containing a yeast integration cassette for RPL18B-driven 
            expression of PEX25-Link-Scarlet(rfp), a marker for the peroxisome.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6827)
  AUTHORS   Bean BDM, Whiteway M, Martin VJJ
  TITLE     The MyLo CRISPR-Cas9 Toolkit: A Markerless Yeast Localization and 
            Overexpression CRISPR-Cas9 Toolkit.
  JOURNAL   G3 (Bethesda). 2022 Jun 16. pii: 6609175. doi: 
            10.1093/g3journal/jkac154.
  PUBMED    35708612
REFERENCE   2  (bases 1 to 6827)
  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). 2022 Jun 16. pii: 6609175. doi: 
            10.1093/g3journal/jkac154."
COMMENT     Sequence Label: pBBK85
            pHomL-RPL18Bp-PEX25-Link-Scarlet(rfp)-S...
FEATURES             Location/Qualifiers
     source          1..6827
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(1..792,793..861))
                     /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     624..643
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(862..966)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1069..1657
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1558..1577
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
       61 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
      121 cagtgctgca atgataccgc gggacccacg ctcaccggct ccagatttat cagcaataaa
      181 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
      241 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
      301 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
      361 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
      421 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
      481 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
      541 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
      601 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
      661 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
      721 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
      781 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
      841 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
      901 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
      961 tccgcgcaca tttccccgaa aagtgccacc tgtcatgacc aaaatccctt aacgtgagtt
     1021 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     1081 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     1141 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     1201 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     1261 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     1321 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     1381 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     1441 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     1501 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     1561 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     1621 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     1681 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     1741 ttctgtggat aaccgtgcgg ccgccaatgt tatctattgg cagatgttca tacggggctt
     1801 tcaaatattg atgaaattat gtgatgttta gaagaagatt cgaactgttt tcagtagatt
     1861 tggtaactgt gcaaccataa ctcatgcccc ctatttatcg gcattgagat tgtaaaggaa
     1921 gtaaaacgca tttttgcaga tctgttctct tatgtatttt taatcgtcct tgtatggaag
     1981 tatcaaaggg gacgttcttc acctccttgg aaaacgcaaa ctttcttttt gcctatcaca
     2041 gtatcttatc gagctaacta ttttcgacac acatgaaaaa gcagaaatat taacgaaaaa
     2101 gaaaagaaag accatgtcat gtacgggcaa tcagaatatg caagatgagc gggaacgggc
     2161 gaaatgtgca cgcgcacaac ttacgaaacg cggatgagtc actgacagcc accgcagagg
     2221 ttctgactcc tactgagctc tattggaggt ggcagaaccg gtaatcccac agtgagcaac
     2281 ttgacaaacc aacagatata ggctgtgtct taaaatatac catccaatac cttgatgaac
     2341 ttttcaaaga tgactaaaat aagtgaaatt tcaacattaa cttcgagctg taaacacagt
     2401 ggtttctttg cataaacacc atcagcctca agtcgtcaag taaagatttc gtgttcatgc
     2461 agatagataa caatctatat gttgataatt agcgttgcct catcaatgcg tacacgaaga
     2521 atatactaaa aaatgagcag gcaagataaa cgaaggcaaa gatgacagag cagaaagccc
     2581 tagtaaagcg tattacaaat gaaaccaaga ttcagattgc gatccctgaa ttcgcatcta
     2641 gatggtagag ccacaaacag ccggtacaag caacgatctc caggaccatc tgaatcatgc
     2701 gcggatgaca cgaactcacg acggcgatca cagacattaa cccacagtac agacactgcg
     2761 acaacgtggc aattcgtcgc aataccgtct cactgaactg gccgataatt gcagacgaac
     2821 gaagaggatg tccaatattt tttttaagga ataaggatac ttcaagacta gattcccccc
     2881 tgcattccca tcagaaccgt aaaccttggc gctttccttg ggaagtattc aagaagtgcc
     2941 ttgtccggtt tctgtggctc acaaaccagc gcgcccgata tggctttctt ttcacttatg
     3001 aatgtaccag tacgggacaa ttagaacgct cctgtaacaa tctctttgca aatgtggggt
     3061 tacattctaa ccatgtcaca ctgctgacga aattcaaagt aaaaaaaaat gggaccacgt
     3121 cttgagaacg atagattttc tttattttac attgaacagt cgttgtctca gcgcgcttta
     3181 tgttttcatt catacttcat attataaaat aacaaaagaa gaatttcata ttcacgccca
     3241 agaaatcagg ctgctttcca aatgcaattg acacttcatt agccatcaca caaaactctt
     3301 tcttgctgga gcttctttta aaaaagacct cagtacacca aacacgttac ccgacctcgt
     3361 tattttacga caactatgat aaaattctga agaaaaaata aaaaaatttt catacttctt
     3421 gcttttattt aaaccattga atgatttctt ttgaacaaaa ctacctgttt caccaaagga
     3481 aatagaaaga aaaaatcaat tagaagaaaa caaaaaacaa aagatctatg agtcagtttg
     3541 gcacgacaga tatcgtttcc ggttcagaaa caccacctta ttctggggca agttaccagg
     3601 acgcccagga tgacaatacc catccacatt catcagacgc cggtgcagag aagttcagtg
     3661 caggcagtgg aagcgagtcg catactgaat cctcgagaag tgatgatgag gattctcaag
     3721 caaaaacaaa gatggtggat aatattacca tcttaaaata tatattggat tctctatctg
     3781 gtagagacaa attagccaaa attatcaaat acgctctgga tatattaaaa ctgtttatcg
     3841 aaaagtctaa gcgaaattta actgttttgg acccttccgt attaacgtac tataccaaga
     3901 ttttgaaaaa tctgactgta aaagtcgcac taaggcatcc aatcacggta attaaagtac
     3961 ttctcctgtc tcttttgaga aattttgaca agaaaattga ttttatcagt caacaattaa
     4021 gtacttttag atatatttta aggtttggcg gcacaccatt ccgagtgtgc agtttcttgg
     4081 ggaaattcaa taaaaccaga aagtgcaact ttcaaattga ccaaattaag aaaatatggt
     4141 ttaatgaagc ttcattgaga gaatttttgg acttgtatta tggtatcttt gatgaattag
     4201 acttgctata caaattgaaa atatggacaa ataaatcatt ctattccttt gtcagtagac
     4261 aagagtctct tgcatggcaa tacgacattc ttttaagctt gaaggaccat tggctcaacc
     4321 tgcaaagctt acagaagaga caattagaat tggaagtgca actaaaagtt caaaataacg
     4381 cgttactact ctcacccata ctgatgcatc aggcccataa agacgatggc tcacaatcgc
     4441 caattaggaa acagttgtta aatgacctaa atgttaataa cgacgcagag gtgttaatac
     4501 acaaacaact aaaggccatc aaggatgaaa agacacttgt ttatttagat attgcaagat
     4561 tatcgtttga ttgcatggct aatacatcag atattctaaa tttaaaaact ccaaagggaa
     4621 cgtatgccgt tctgtccctg ggatccggct taactggact ggttaaactt tggataacaa
     4681 caaagaggtc actttgctct tcaaaagatg gatccggtcc ttctggccca actggcatgg
     4741 ttagtaaagg tgaagctgtt ataaaagaat tcatgaggtt taaagttcat atggaaggtt
     4801 caatgaatgg tcatgaattt gaaattgaag gtgaaggtga aggtagacca tatgaaggta
     4861 cacaaactgc taaattgaaa gttactaaag gtggtccatt accattttct tgggatattt
     4921 tgtctccaca attcatgtat ggttctagag cttttattaa gcatccagct gatattccag
     4981 attattataa acaatctttt cctgaaggtt ttaaatggga aagagttatg aatttcgaag
     5041 atggtggtgc tgttactgtt actcaagata cttctttgga agatggtact ttaatctata
     5101 aagttaaatt gagaggtact aattttccac cagatggtcc agttatgcaa aagaaaacta
     5161 tgggttggga agcatctact gaaagattgt atccagaaga tggtgttttg aaaggagaca
     5221 ttaagatggc tttgagattg aaagatggtg gtcggtacct ggccgacttc aagaccacct
     5281 ataaagctaa aaaaccagtt caaatgccag gtgcatataa tgttgataga aagttagata
     5341 taacgtcgca taacgaggac tatactgtag ttgaacaata tgaacgtagt gaaggtagac
     5401 atagtaccgg aggaatggat gaattgtata aatagctcga gtggcgccaa ttggtgcggc
     5461 aattgataat aacgaaaatg tcttttaatg atctgggtat aatgaggaat tttccgaacg
     5521 tttttacttt atatatatat atacatgtaa catatattct atacgctata gagaaaggaa
     5581 atttttcaat taaaaaaaaa atagagaaag agtttcactt cttgattatc gctaacacta
     5641 atggttgaag tactgctact ttaattttat gctgccaatg agacgacggg gtcatcacgg
     5701 ctcatcatgc gccaaacaaa tgtgtgcaat acacgctcgg atgactgcat gatgaccgca
     5761 ctgactgggg acagcagatc cacctaagcc tgtgagagaa gcagacaccc gacagatcaa
     5821 ggcagttaac tagtgcactg cagtacaaat cgtccccaac aaaagtgggc tctcaaaatt
     5881 catcacattt aaatgcatat aggaagagca acagttggtt tgcatctgat gttccttaaa
     5941 gatttcgaca taatgtgcga agtagatccc ttcagcactg tacttgtttt tgcgactaga
     6001 ttgtaaatca ttctttattt aatctctttc tttaactact gcttaaagta taatttggtc
     6061 cgtagtttaa taactatact aagcgaacgc tctctcgaaa tttttcttaa cgcgtccttg
     6121 tactgcgtct aacgcttttg ccacttggat ttctattata ggaaatagtc tcacttactg
     6181 ggcgacgaat tttcgcgttt tgatgaagca caggaagaat ttctttatgc tcagaagttt
     6241 gacagcaagc aagttcatca ttcgaactag ccttattgtt ttagttcagt gacagcgaac
     6301 tgccgtactc gatgctttat ttctcacggt agagcggaag aacagataga tccctacata
     6361 ttgctaccac ttctattaca caatagtttc aatagcttgc agcgtagcta aactctaaaa
     6421 tttatctaaa tcactcatat aaaccgaacc cttccccttc cgcttatagt acagctgatg
     6481 ctccttcact attttaacat gtggaattct tgaaagaatg aaatcgccat gccaagccat
     6541 cacacggtct tttatgcaat tgattgaccg cctgcaacac ataggcagta aaatttttac
     6601 aagcagtagc agaacaggcc acacaatcgc aagtgattaa cgtccacaca ggtatagggt
     6661 ttctggacca tatgatacat gctctggcca agcattccgg ctggtcgcta atcgttgagt
     6721 gccgagcggc cgcgattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     6781 agttttaaat caatctaaag tatatatgag taaacttggt ctgacag
//
