LOCUS       sequence-360951-        7496 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vector containing a yeast integration cassette for RPL18B-driven 
            expression of TOM70-Link-Scarlet(rfp), a marker for the 
            mitochondria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7496)
  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 7496)
  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: pBBK87
            pHomL-RPL18Bp-TOM70-Link-Scarlet(rfp)-S...
FEATURES             Location/Qualifiers
     source          1..7496
                     /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 aagagcttca
     3541 ttacaaggaa caagacagcc attttggcaa ccgttgctgc tacaggtact gccatcggtg
     3601 cctactatta ttacaaccaa ttgcaacaac aacaacaacg aggaaaaaag aacacgatca
     3661 acaaagatga aaaaaaggac acaaaggact ctcaaaagga gactgaaggt gctaagaaat
     3721 ctacagcccc atcaaatcct cctatctacc cggtttctag taatggcgaa ccagattttt
     3781 ccaataaggc aaatttcacc gctgaagaaa aggataaata tgcattagcg ttaaaggaca
     3841 aaggtaacca gttctttaga aataaaaaat atgacgatgc tattaagtac tataattggg
     3901 cattagaatt gaaagaagac ccagttttct actcgaattt atcggcttgc tatgtttctg
     3961 tgggtgactt gaaaaaagtt gttgaaatga gtactaaggc tcttgaatta aaaccagact
     4021 actcaaaagt tttactgaga agagcttctg ctaacgaagg gctaggaaaa tttgcggatg
     4081 cgatgttcga tttgtctgta ttgtctctaa atggtgactt caatgacgcc tctattgaac
     4141 caatgttgga gaggaacttg aataagcaag ctatgtctaa attgaaagaa aagtttggcg
     4201 atattgacac cgctactgct actccaactg aattatccac ccaaccagct aaagaacgca
     4261 aagacaagca ggaaaacttg ccttcagtta catccatggc ctctttcttt ggtattttca
     4321 aacccgagtt gacttttgcc aattatgatg aatccaatga agctgataaa gagttaatga
     4381 acggtttaag taatttgtac aaaagatctc ccgaaagtta cgacaaggct gatgagtctt
     4441 tcacgaaagc tgcaaggtta tttgaagaac aattggacaa gaacaatgag gatgaaaagt
     4501 taaaggaaaa attggccatc tcattagaac acacaggtat tttcaagttc ttgaaaaacg
     4561 atccattggg agctcacgaa gacatcaaga aagctattga attgttccca agagtcaatt
     4621 catatatcta catggcatta atcatggctg atagaaatga ctcgactgaa tattataact
     4681 acttcgataa agcgctgaaa ctggactcaa acaattcttc tgtttactat caccgcggcc
     4741 aaatgaattt cattttacaa aactatgatc aagcaggaaa agattttgat aaagctaagg
     4801 aattagaccc agagaatatc ttcccttata ttcaattagc atgtctagcg taccgtgaaa
     4861 acaagtttga tgactgtgaa actttgttta gtgaagctaa gagaaaattt ccagaggcac
     4921 cagaagttcc aaatttcttt gctgaaattt taacagacaa gaacgatttt gacaaggctt
     4981 tgaagcagta cgatttggct attgaattag aaaacaagtt ggatggtatt tatgttggaa
     5041 ttgcgccttt ggtcggtaaa gccactttgt tgacaagaaa tccaacagta gaaaatttta
     5101 ttgaagccac gaatttatta gaaaaagcgt ccaaactaga cccaagaagt gagcaagcta
     5161 aaatcggttt agctcaaatg aaattgcagc aggaagatat cgatgaagct attacattat
     5221 tcgaagaatc cgctgatttg gctagaacta tggaagagaa attacaagcc attacttttg
     5281 ctgaagccgc taaagttcaa caaagaatta ggtctgatcc agtattggct aaaaagattc
     5341 aagaaacttt agctaaatta cgcgaacagg gtttaatggg atccggtcct tctggcccaa
     5401 ctggcatggt tagtaaaggt gaagctgtta taaaagaatt catgaggttt aaagttcata
     5461 tggaaggttc aatgaatggt catgaatttg aaattgaagg tgaaggtgaa ggtagaccat
     5521 atgaaggtac acaaactgct aaattgaaag ttactaaagg tggtccatta ccattttctt
     5581 gggatatttt gtctccacaa ttcatgtatg gttctagagc ttttattaag catccagctg
     5641 atattccaga ttattataaa caatcttttc ctgaaggttt taaatgggaa agagttatga
     5701 atttcgaaga tggtggtgct gttactgtta ctcaagatac ttctttggaa gatggtactt
     5761 taatctataa agttaaattg agaggtacta attttccacc agatggtcca gttatgcaaa
     5821 agaaaactat gggttgggaa gcatctactg aaagattgta tccagaagat ggtgttttga
     5881 aaggagacat taagatggct ttgagattga aagatggtgg tcggtacctg gccgacttca
     5941 agaccaccta taaagctaaa aaaccagttc aaatgccagg tgcatataat gttgatagaa
     6001 agttagatat aacgtcgcat aacgaggact atactgtagt tgaacaatat gaacgtagtg
     6061 aaggtagaca tagtaccgga ggaatggatg aattgtataa atagctcgag tggcgccaat
     6121 tggtgcggca attgataata acgaaaatgt cttttaatga tctgggtata atgaggaatt
     6181 ttccgaacgt ttttacttta tatatatata tacatgtaac atatattcta tacgctatag
     6241 agaaaggaaa tttttcaatt aaaaaaaaaa tagagaaaga gtttcacttc ttgattatcg
     6301 ctaacactaa tggttgaagt actgctactt taattttatg ctgccaatga gacgacgggg
     6361 tcatcacggc tcatcatgcg ccaaacaaat gtgtgcaata cacgctcgga tgactgcatg
     6421 atgaccgcac tgactgggga cagcagatcc acctaagcct gtgagagaag cagacacccg
     6481 acagatcaag gcagttaact agtgcactgc agtacaaatc gtccccaaca aaagtgggct
     6541 ctcaaaattc atcacattta aatgcatata ggaagagcaa cagttggttt gcatctgatg
     6601 ttccttaaag atttcgacat aatgtgcgaa gtagatccct tcagcactgt acttgttttt
     6661 gcgactagat tgtaaatcat tctttattta atctctttct ttaactactg cttaaagtat
     6721 aatttggtcc gtagtttaat aactatacta agcgaacgct ctctcgaaat ttttcttaac
     6781 gcgtccttgt actgcgtcta acgcttttgc cacttggatt tctattatag gaaatagtct
     6841 cacttactgg gcgacgaatt ttcgcgtttt gatgaagcac aggaagaatt tctttatgct
     6901 cagaagtttg acagcaagca agttcatcat tcgaactagc cttattgttt tagttcagtg
     6961 acagcgaact gccgtactcg atgctttatt tctcacggta gagcggaaga acagatagat
     7021 ccctacatat tgctaccact tctattacac aatagtttca atagcttgca gcgtagctaa
     7081 actctaaaat ttatctaaat cactcatata aaccgaaccc ttccccttcc gcttatagta
     7141 cagctgatgc tccttcacta ttttaacatg tggaattctt gaaagaatga aatcgccatg
     7201 ccaagccatc acacggtctt ttatgcaatt gattgaccgc ctgcaacaca taggcagtaa
     7261 aatttttaca agcagtagca gaacaggcca cacaatcgca agtgattaac gtccacacag
     7321 gtatagggtt tctggaccat atgatacatg ctctggccaa gcattccggc tggtcgctaa
     7381 tcgttgagtg ccgagcggcc gcgattatca aaaaggatct tcacctagat ccttttaaat
     7441 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacag
//
