LOCUS       sequence-360782-        6746 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vector containing a yeast integration cassette for RPL18B-driven 
            expression of SPC42-Link-Neon(gfp), a marker for the spindle pole.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6746)
  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 6746)
  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: pBBK48 pHomL-RPL18Bp-SPC42-Link-Neon(gfp)-SSA1...
FEATURES             Location/Qualifiers
     source          1..6746
                     /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"
     regulatory      4899..4908
                     /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 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 ggcatggttt ctaagggtga agaagataac atggcttctt
     4681 tgccagccac ccacgagttg cacatcttcg gttccattaa cggtgtcgac ttcgatatgg
     4741 tcggtcaagg taccggtaac ccaaacgacg gttacgaaga attgaacttg aagtctacca
     4801 agggcgactt gcagttctcc ccttggattt tggttccaca catcggttac ggtttccacc
     4861 aatacttgcc atacccagac ggtatgtccc cattccaagc cgccatggtt gacggttctg
     4921 gttaccaagt ccacagaacc atgcaattcg aagatggtgc ttccttgacc gtgaactaca
     4981 gatacaccta cgaaggttct cacatcaagg gcgaagccca agttaagggt accggtttcc
     5041 ctgctgacgg tccagtcatg actaactcct tgaccgctgc tgactggtgt cgttccaaga
     5101 aaacctaccc aaacgataag accatcattt ccactttcaa gtggtcctac accaccggta
     5161 acggcaagag ataccgctct accgctcgta ccacctacac cttcgccaag ccaatggctg
     5221 ccaactactt gaagaaccag cctatgtacg ttttccgtaa gaccgagctg aagcattcca
     5281 agactgaatt gaacttcaag gaatggcaaa aggctttcac cgacgtcatg ggtatggacg
     5341 aattgtacaa gtaactcgag tggcgccaat tggtgcggca attgataata acgaaaatgt
     5401 cttttaatga tctgggtata atgaggaatt ttccgaacgt ttttacttta tatatatata
     5461 tacatgtaac atatattcta tacgctatag agaaaggaaa tttttcaatt aaaaaaaaaa
     5521 tagagaaaga gtttcacttc ttgattatcg ctaacactaa tggttgaagt actgctactt
     5581 taattttatg ctgccaatga gacgacgggg tcatcacggc tcatcatgcg ccaaacaaat
     5641 gtgtgcaata cacgctcgga tgactgcatg atgaccgcac tgactgggga cagcagatcc
     5701 acctaagcct gtgagagaag cagacacccg acagatcaag gcagttaact agtgcactgc
     5761 agtacaaatc gtccccaaca aaagtgggct ctcaaaattc atcacattta aatgcatata
     5821 ggaagagcaa cagttggttt gcatctgatg ttccttaaag atttcgacat aatgtgcgaa
     5881 gtagatccct tcagcactgt acttgttttt gcgactagat tgtaaatcat tctttattta
     5941 atctctttct ttaactactg cttaaagtat aatttggtcc gtagtttaat aactatacta
     6001 agcgaacgct ctctcgaaat ttttcttaac gcgtccttgt actgcgtcta acgcttttgc
     6061 cacttggatt tctattatag gaaatagtct cacttactgg gcgacgaatt ttcgcgtttt
     6121 gatgaagcac aggaagaatt tctttatgct cagaagtttg acagcaagca agttcatcat
     6181 tcgaactagc cttattgttt tagttcagtg acagcgaact gccgtactcg atgctttatt
     6241 tctcacggta gagcggaaga acagatagat ccctacatat tgctaccact tctattacac
     6301 aatagtttca atagcttgca gcgtagctaa actctaaaat ttatctaaat cactcatata
     6361 aaccgaaccc ttccccttcc gcttatagta cagctgatgc tccttcacta ttttaacatg
     6421 tggaattctt gaaagaatga aatcgccatg ccaagccatc acacggtctt ttatgcaatt
     6481 gattgaccgc ctgcaacaca taggcagtaa aatttttaca agcagtagca gaacaggcca
     6541 cacaatcgca agtgattaac gtccacacag gtatagggtt tctggaccat atgatacatg
     6601 ctctggccaa gcattccggc tggtcgctaa tcgttgagtg ccgagcggcc gcgattatca
     6661 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     6721 atatatgagt aaacttggtc tgacag
//
