LOCUS       sequence-360953-        6524 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vector containing a yeast integration cassette for expressing 
            TDH3p-Neon(gfp)-Link-2xPH(PLCd), an overexpressed (bright) marker 
            for the plasma membrane.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6524)
  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 6524)
  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: pBBK88
            pHomL-TDH3p-Neon(gfp)-Link-2xPH(PLCd)-S...
FEATURES             Location/Qualifiers
     source          1..6524
                     /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"
     promoter        2834..3500
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     3370..3393
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     regulatory      3762..3771
                     /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 catgaaaaag cagaaatatt aacgaaaaag
     2101 aaaagaaaga ccatgtcatg tacgggcaat cagaatatgc aagatgagcg ggaacgggcg
     2161 aaatgtgcac gcgcacaact tacgaaacgc ggatgagtca ctgacagcca ccgcagaggt
     2221 tctgactcct actgagctct attggaggtg gcagaaccgg taatcccaca gtgagcaact
     2281 tgacaaacca acagatatag gctgtgtctt aaaatatacc atccaatacc ttgatgaact
     2341 tttcaaagat gactaaaata agtgaaattt caacattaac ttcgagctgt aaacacagtg
     2401 gtttctttgc ataaacacca tcagcctcaa gtcgtcaagt aaagatttcg tgttcatgca
     2461 gatagataac aatctatatg ttgataatta gcgttgcctc atcaatgcgt acacgaagaa
     2521 tatactaaaa aatgagcagg caagataaac gaaggcaaag atgacagagc agaaagccct
     2581 agtaaagcgt attacaaatg aaaccaagat tcagattgcg atccctgaat tcgcatctag
     2641 atggtagagc cacaaacagc cggtacaagc aacgatctcc aggaccatct gaatcatgcg
     2701 cggatgacac gaactcacga cggcgatcac agacattaac ccacagtaca gacactgcga
     2761 caacgtggca attcgtcgca ataccgtctc actgaactgg ccgataattg cagacgaacg
     2821 cagttcgagt ttatcattat caatactgcc atttcaaaga atacgtaaat aattaatagt
     2881 agtgattttc ctaactttat ttagtcaaaa aattagcctt ttaattctgc tgtaacccgt
     2941 acatgcccaa aatagggggc gggttacaca gaatatataa catcgtaggt gtctgggtga
     3001 acagtttatt cctggcatcc actaaatata atggagcccg ctttttaagc tggcatccag
     3061 aaaaaaaaag aatcccagca ccaaaatatt gttttcttca ccaaccatca gttcataggt
     3121 ccattctctt agcgcaacta cagagaacag gggcacaaac aggcaaaaaa cgggcacaac
     3181 ctcaatggag tgatgcaacc tgcctggagt aaatgatgac acaaggcaat tgacccacgc
     3241 atgtatctat ctcattttct tacaccttct attaccttct gctctctctg atttggaaaa
     3301 agctgaaaaa aaaggttgaa accagttccc tgaaattatt cccctacttg actaataagt
     3361 atataaagac ggtaggtatt gattgtaatt ctgtaaatct atttcttaaa cttcttaaat
     3421 tctactttta tagttagtct tttttttagt tttaaaacac caagaactta gtttcgaata
     3481 aacacacata aacaaacaaa agatctatgg tttctaaggg tgaagaagat aacatggctt
     3541 ctttgccagc cacccacgag ttgcacatct tcggttccat taacggtgtc gacttcgata
     3601 tggtcggtca aggtaccggt aacccaaacg acggttacga agaattgaac ttgaagtcta
     3661 ccaagggcga cttgcagttc tccccttgga ttttggttcc acacatcggt tacggtttcc
     3721 accaatactt gccataccca gacggtatgt ccccattcca agccgccatg gttgacggtt
     3781 ctggttacca agtccacaga accatgcaat tcgaagatgg tgcttccttg accgtgaact
     3841 acagatacac ctacgaaggt tctcacatca agggcgaagc ccaagttaag ggtaccggtt
     3901 tccctgctga cggtccagtc atgactaact ccttgaccgc tgctgactgg tgtcgttcca
     3961 agaaaaccta cccaaacgat aagaccatca tttccacttt caagtggtcc tacaccaccg
     4021 gtaacggcaa gagataccgc tctaccgctc gtaccaccta caccttcgcc aagccaatgg
     4081 ctgccaacta cttgaagaac cagcctatgt acgttttccg taagaccgag ctgaagcatt
     4141 ccaagactga attgaacttc aaggaatggc aaaaggcttt caccgacgtc atgggtatgg
     4201 acgaattgta caagtctggt ccttctggcc caactggttc tccaggtaga actctcagat
     4261 cccacgggct ccaggatgac ccggaccttc aggcccttct gaagggcagc cagcttctga
     4321 aggtgaagtc cagctcgtgg cgtagggaac gcttctacaa gctacaggag gactgcaaga
     4381 ccatctggca ggaatctcga aaggtcatga ggtccccgga gtcgcagctg ttctccatcg
     4441 aggacattca ggaggtacgg atgggacacc gcacagaagg cctggagaag tttgcccgag
     4501 acatccccga ggatcgatgc ttctccattg tcttcaagga ccagcgcaac accctagacc
     4561 tcattgcccc atcaccagct gacgctcagc actgggtgca gggcctgcgc aagatcatcc
     4621 accactccgg ctccatggac cagcggcaga aggcagcggc agcaggtggc gcattcagaa
     4681 agagcgaatt cgcccttgcg gaatccgccg cggcagctca cgggctccag gatgacccgg
     4741 accttcaggc ccttctgaag ggcagccagc ttctgaaggt gaagtccagc tcgtggcgta
     4801 gggaacgctt ctacaagcta caggaggact gcaagaccat ctggcaggaa tctcgaaagg
     4861 tcatgaggtc cccggagtcg cagctgttct ccatcgagga cattcaggag gtacggatgg
     4921 gacaccgcac agaaggcctg gagaagtttg cccgagacat ccccgaggat cgatgcttct
     4981 ccattgtctt caaggaccag cgcaacaccc tagacctcat tgccccatca ccagctgacg
     5041 ctcagcactg ggtgcagggc ctgcgcaaga tcatccacca ctccggctcc atggaccagc
     5101 ggcagaagta gtcgacctcg gtcgccgaga tcctaactcg aggccaattg gtgcggcaat
     5161 tgataataac gaaaatgtct tttaatgatc tgggtataat gaggaatttt ccgaacgttt
     5221 ttactttata tatatatata catgtaacat atattctata cgctatagag aaaggaaatt
     5281 tttcaattaa aaaaaaaata gagaaagagt ttcacttctt gattatcgct aacactaatg
     5341 gttgaagtac tgctacttta attttatgct gccaatgaga cgacggggtc atcacggctc
     5401 atcatgcgcc aaacaaatgt gtgcaataca cgctcggatg actgcatgat gaccgcactg
     5461 actggggaca gcagatccac ctaagcctgt gagagaagca gacacccgac agatcaaggc
     5521 agttaactag tgcactgcag tacaaatcgt ccccaacaaa agtgggctct caaaattcat
     5581 cacatttaaa tgcatatagg aagagcaaca gttggtttgc atctgatgtt ccttaaagat
     5641 ttcgacataa tgtgcgaagt agatcccttc agcactgtac ttgtttttgc gactagattg
     5701 taaatcattc tttatttaat ctctttcttt aactactgct taaagtataa tttggtccgt
     5761 agtttaataa ctatactaag cgaacgctct ctcgaaattt ttcttaacgc gtccttgtac
     5821 tgcgtctaac gcttttgcca cttggatttc tattatagga aatagtctca cttactgggc
     5881 gacgaatttt cgcgttttga tgaagcacag gaagaatttc tttatgctca gaagtttgac
     5941 agcaagcaag ttcatcattc gaactagcct tattgtttta gttcagtgac agcgaactgc
     6001 cgtactcgat gctttatttc tcacggtaga gcggaagaac agatagatcc ctacatattg
     6061 ctaccacttc tattacacaa tagtttcaat agcttgcagc gtagctaaac tctaaaattt
     6121 atctaaatca ctcatataaa ccgaaccctt ccccttccgc ttatagtaca gctgatgctc
     6181 cttcactatt ttaacatgtg gaattcttga aagaatgaaa tcgccatgcc aagccatcac
     6241 acggtctttt atgcaattga ttgaccgcct gcaacacata ggcagtaaaa tttttacaag
     6301 cagtagcaga acaggccaca caatcgcaag tgattaacgt ccacacaggt atagggtttc
     6361 tggaccatat gatacatgct ctggccaagc attccggctg gtcgctaatc gttgagtgcc
     6421 gagcggccgc gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     6481 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acag
//
