LOCUS       sequence-360844-        5738 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vector containing a yeast integration cassette for RPL18B-driven 
            expression of preCOX4-Link-Neon(gfp), a marker for the mitochondria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5738)
  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 5738)
  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: pBBK65
            pHomL-RPL18Bp-preCOX4-Link-Neon(gfp)-SS...
FEATURES             Location/Qualifiers
     source          1..5738
                     /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"
     CDS             3528..3593
                     /codon_start=1
                     /product="mitochondrial presequence of yeast cytochrome c 
                     oxidase subunit IV (Hurt et al., 1984)"
                     /label=pre-CoxIV
                     /translation="MLSLRQSIRFFKPATRTLCSSR"
     regulatory      3891..3900
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
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 ctttcactac
     3541 gtcaatctat aagatttttc aagccagcca caagaacttt gtgtagctct agatatctgc
     3601 ttcagcaagg atccggtcct tctggcccaa ctggcatggt ttctaagggt gaagaagata
     3661 acatggcttc tttgccagcc acccacgagt tgcacatctt cggttccatt aacggtgtcg
     3721 acttcgatat ggtcggtcaa ggtaccggta acccaaacga cggttacgaa gaattgaact
     3781 tgaagtctac caagggcgac ttgcagttct ccccttggat tttggttcca cacatcggtt
     3841 acggtttcca ccaatacttg ccatacccag acggtatgtc cccattccaa gccgccatgg
     3901 ttgacggttc tggttaccaa gtccacagaa ccatgcaatt cgaagatggt gcttccttga
     3961 ccgtgaacta cagatacacc tacgaaggtt ctcacatcaa gggcgaagcc caagttaagg
     4021 gtaccggttt ccctgctgac ggtccagtca tgactaactc cttgaccgct gctgactggt
     4081 gtcgttccaa gaaaacctac ccaaacgata agaccatcat ttccactttc aagtggtcct
     4141 acaccaccgg taacggcaag agataccgct ctaccgctcg taccacctac accttcgcca
     4201 agccaatggc tgccaactac ttgaagaacc agcctatgta cgttttccgt aagaccgagc
     4261 tgaagcattc caagactgaa ttgaacttca aggaatggca aaaggctttc accgacgtca
     4321 tgggtatgga cgaattgtac aagtaactcg agtggcgcca attggtgcgg caattgataa
     4381 taacgaaaat gtcttttaat gatctgggta taatgaggaa ttttccgaac gtttttactt
     4441 tatatatata tatacatgta acatatattc tatacgctat agagaaagga aatttttcaa
     4501 ttaaaaaaaa aatagagaaa gagtttcact tcttgattat cgctaacact aatggttgaa
     4561 gtactgctac tttaatttta tgctgccaat gagacgacgg ggtcatcacg gctcatcatg
     4621 cgccaaacaa atgtgtgcaa tacacgctcg gatgactgca tgatgaccgc actgactggg
     4681 gacagcagat ccacctaagc ctgtgagaga agcagacacc cgacagatca aggcagttaa
     4741 ctagtgcact gcagtacaaa tcgtccccaa caaaagtggg ctctcaaaat tcatcacatt
     4801 taaatgcata taggaagagc aacagttggt ttgcatctga tgttccttaa agatttcgac
     4861 ataatgtgcg aagtagatcc cttcagcact gtacttgttt ttgcgactag attgtaaatc
     4921 attctttatt taatctcttt ctttaactac tgcttaaagt ataatttggt ccgtagttta
     4981 ataactatac taagcgaacg ctctctcgaa atttttctta acgcgtcctt gtactgcgtc
     5041 taacgctttt gccacttgga tttctattat aggaaatagt ctcacttact gggcgacgaa
     5101 ttttcgcgtt ttgatgaagc acaggaagaa tttctttatg ctcagaagtt tgacagcaag
     5161 caagttcatc attcgaacta gccttattgt tttagttcag tgacagcgaa ctgccgtact
     5221 cgatgcttta tttctcacgg tagagcggaa gaacagatag atccctacat attgctacca
     5281 cttctattac acaatagttt caatagcttg cagcgtagct aaactctaaa atttatctaa
     5341 atcactcata taaaccgaac ccttcccctt ccgcttatag tacagctgat gctccttcac
     5401 tattttaaca tgtggaattc ttgaaagaat gaaatcgcca tgccaagcca tcacacggtc
     5461 ttttatgcaa ttgattgacc gcctgcaaca cataggcagt aaaattttta caagcagtag
     5521 cagaacaggc cacacaatcg caagtgatta acgtccacac aggtataggg tttctggacc
     5581 atatgataca tgctctggcc aagcattccg gctggtcgct aatcgttgag tgccgagcgg
     5641 ccgcgattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     5701 tcaatctaaa gtatatatga gtaaacttgg tctgacag
//
