LOCUS       sequence-360852-        6734 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vector containing a yeast integration cassette for RPL18B-driven 
            expression of SPC42-Link-Scarlet(rfp), a marker for the spindle 
            pole.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6734)
  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 6734)
  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: pBBK69
            pHomL-RPL18Bp-SPC42-Link-Scarlet(rfp)-S...
FEATURES             Location/Qualifiers
     source          1..6734
                     /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 aacggatctc
     3541 ccactcctaa gcgatattca tccaaatctt caaggcttta cgacgactac tataacatcc
     3601 cgtatcaata ttccaatccg actccaatga atagagacta taatgacgtg gggtcacgta
     3661 tcaatgcaga taaactggta ccagaggaat acaaacggaa tacagaattt atcaacaaag
     3721 ctgtccaaca aaataaagaa cttaacttca aactgagaga aaagcagaac gaaatattcg
     3781 aactcaagaa gattgcagaa actctgcgct caaagctcga aaaatatgtt gacattacca
     3841 aaaagcttga agaccaaaac ctcaacctac agataaaaat aagcgaccta gaaaagaaac
     3901 tttctgacgc taattctact ttcaaagaaa tgagattccc caaggtcaag gaccccatgg
     3961 tagatgacga ccctgtctcc gaaaattatg atcagataaa tgtaccaaaa cacagagcac
     4021 cggatgctac aggtaatcct cgtacaacta acaaagtctc caatacttca gaccaagata
     4081 gcagactaaa agccattgaa agaaccctct cggtcctcac aaattatgtt atgaggtcag
     4141 aagacggtaa taacgacagg atgtcaccct taccttctcc tttaaatacg atattgccaa
     4201 taaataatcg tctgaatttt caagagccaa agagatataa tccaaccgtt aaggtaaatc
     4261 cgtctgatga cgatatcatg atgtatgaaa gtgcagagct gaagcgtgtc gaagaagaaa
     4321 tagaggagct aaagagaaaa atacttgtta ggaaaaaaca tgacttaaga aaattgtctt
     4381 taaataacca actccaagag ctgcaaagca tgatggacgg tgatgacaat ataaaacttg
     4441 acaatgtttc gaagcataat catgctacgc atcgtcattc ttcccaatcc agtcgcgatt
     4501 actcaccctc ttcggatgct tgcttggagt gttctaatga tttatacgag aaaaatagag
     4561 tcaagcctga aaataatatg tcagaaacat tcgcaactcc cactcccaat aatcgaggat
     4621 ccggtccttc tggcccaact ggcatggtta gtaaaggtga agctgttata aaagaattca
     4681 tgaggtttaa agttcatatg gaaggttcaa tgaatggtca tgaatttgaa attgaaggtg
     4741 aaggtgaagg tagaccatat gaaggtacac aaactgctaa attgaaagtt actaaaggtg
     4801 gtccattacc attttcttgg gatattttgt ctccacaatt catgtatggt tctagagctt
     4861 ttattaagca tccagctgat attccagatt attataaaca atcttttcct gaaggtttta
     4921 aatgggaaag agttatgaat ttcgaagatg gtggtgctgt tactgttact caagatactt
     4981 ctttggaaga tggtacttta atctataaag ttaaattgag aggtactaat tttccaccag
     5041 atggtccagt tatgcaaaag aaaactatgg gttgggaagc atctactgaa agattgtatc
     5101 cagaagatgg tgttttgaaa ggagacatta agatggcttt gagattgaaa gatggtggtc
     5161 ggtacctggc cgacttcaag accacctata aagctaaaaa accagttcaa atgccaggtg
     5221 catataatgt tgatagaaag ttagatataa cgtcgcataa cgaggactat actgtagttg
     5281 aacaatatga acgtagtgaa ggtagacata gtaccggagg aatggatgaa ttgtataaat
     5341 agctcgagtg gcgccaattg gtgcggcaat tgataataac gaaaatgtct tttaatgatc
     5401 tgggtataat gaggaatttt ccgaacgttt ttactttata tatatatata catgtaacat
     5461 atattctata cgctatagag aaaggaaatt tttcaattaa aaaaaaaata gagaaagagt
     5521 ttcacttctt gattatcgct aacactaatg gttgaagtac tgctacttta attttatgct
     5581 gccaatgaga cgacggggtc atcacggctc atcatgcgcc aaacaaatgt gtgcaataca
     5641 cgctcggatg actgcatgat gaccgcactg actggggaca gcagatccac ctaagcctgt
     5701 gagagaagca gacacccgac agatcaaggc agttaactag tgcactgcag tacaaatcgt
     5761 ccccaacaaa agtgggctct caaaattcat cacatttaaa tgcatatagg aagagcaaca
     5821 gttggtttgc atctgatgtt ccttaaagat ttcgacataa tgtgcgaagt agatcccttc
     5881 agcactgtac ttgtttttgc gactagattg taaatcattc tttatttaat ctctttcttt
     5941 aactactgct taaagtataa tttggtccgt agtttaataa ctatactaag cgaacgctct
     6001 ctcgaaattt ttcttaacgc gtccttgtac tgcgtctaac gcttttgcca cttggatttc
     6061 tattatagga aatagtctca cttactgggc gacgaatttt cgcgttttga tgaagcacag
     6121 gaagaatttc tttatgctca gaagtttgac agcaagcaag ttcatcattc gaactagcct
     6181 tattgtttta gttcagtgac agcgaactgc cgtactcgat gctttatttc tcacggtaga
     6241 gcggaagaac agatagatcc ctacatattg ctaccacttc tattacacaa tagtttcaat
     6301 agcttgcagc gtagctaaac tctaaaattt atctaaatca ctcatataaa ccgaaccctt
     6361 ccccttccgc ttatagtaca gctgatgctc cttcactatt ttaacatgtg gaattcttga
     6421 aagaatgaaa tcgccatgcc aagccatcac acggtctttt atgcaattga ttgaccgcct
     6481 gcaacacata ggcagtaaaa tttttacaag cagtagcaga acaggccaca caatcgcaag
     6541 tgattaacgt ccacacaggt atagggtttc tggaccatat gatacatgct ctggccaagc
     6601 attccggctg gtcgctaatc gttgagtgcc gagcggccgc gattatcaaa aaggatcttc
     6661 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     6721 acttggtctg acag
//
