LOCUS       sequence-360826-        6674 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vector containing a yeast integration cassette for RPL18B-driven 
            expression of PIL1-Link-Neon(gfp), a marker for the eisosomes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6674)
  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 6674)
  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: pBBK55 pHomL-RPL18Bp-PIL1-Link-Neon(gfp)-SSA1t...
FEATURES             Location/Qualifiers
     source          1..6674
                     /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      4827..4836
                     /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 cacagaactt
     3541 actctttaag aaattccagg gcacctaccg cctctcaatt acagaaccca ccgccaccac
     3601 catctacaac caaaggtaga ttctttggga agggtggtct agcttacagc tttaggagaa
     3661 gtgctgctgg agcttttggc ccagaattat ccagaaagtt gtctcaattg gttaagattg
     3721 aaaagaatgt tttgaggtcc atggaattga cagccaacga aagacgtgac gctgctaagc
     3781 aattgtctat ttgggggttg gaaaacgatg acgatgtttc cgacatcact gataaattag
     3841 gtgtcttgat ctatgaagtt agtgaattag acgaccaatt tatcgatcgt tatgaccaat
     3901 acagattgac tctaaagtcc atcagagata tcgaaggttc tgttcaacca tctagggaca
     3961 gtaaggacaa gatcaccgac aaaatcgcct acttgaaata caaagatcct caatcaccta
     4021 agattgaggt cttggaacaa gaattggtgc gtgctgaggc tgaatctttg gtcgctgaag
     4081 ctcaattatc taatatcaca aggtcaaagt tgagagctgc tttcaactac caatttgact
     4141 ccatcatcga acattcagag aaaattgctt taatcgctgg ttacggtaag gctctcttgg
     4201 aactattgga cgactctcct gtcactccag gtgaaaccag gcctgcttac gatgggtatg
     4261 aagcctctaa acaaatcatt attgatgctg aaagcgcact gaatgaatgg acactagact
     4321 ctgcccaagt caagcctact ttaagtttca agcaggatta cgaagacttc gaacctgaag
     4381 aaggcgaaga agaggaagag gaagacggtc aaggcaggtg gtccgaagac gaacaagaag
     4441 atggacaaat tgaagaacct gaacaagaag aagaaggtgc tgttgaagaa catgaacaag
     4501 tcggacacca gcaaagtgag tctcttcccc aacaaacaac agctggatcc ggtccttctg
     4561 gcccaactgg catggtttct aagggtgaag aagataacat ggcttctttg ccagccaccc
     4621 acgagttgca catcttcggt tccattaacg gtgtcgactt cgatatggtc ggtcaaggta
     4681 ccggtaaccc aaacgacggt tacgaagaat tgaacttgaa gtctaccaag ggcgacttgc
     4741 agttctcccc ttggattttg gttccacaca tcggttacgg tttccaccaa tacttgccat
     4801 acccagacgg tatgtcccca ttccaagccg ccatggttga cggttctggt taccaagtcc
     4861 acagaaccat gcaattcgaa gatggtgctt ccttgaccgt gaactacaga tacacctacg
     4921 aaggttctca catcaagggc gaagcccaag ttaagggtac cggtttccct gctgacggtc
     4981 cagtcatgac taactccttg accgctgctg actggtgtcg ttccaagaaa acctacccaa
     5041 acgataagac catcatttcc actttcaagt ggtcctacac caccggtaac ggcaagagat
     5101 accgctctac cgctcgtacc acctacacct tcgccaagcc aatggctgcc aactacttga
     5161 agaaccagcc tatgtacgtt ttccgtaaga ccgagctgaa gcattccaag actgaattga
     5221 acttcaagga atggcaaaag gctttcaccg acgtcatggg tatggacgaa ttgtacaagt
     5281 aactcgagtg gcgccaattg gtgcggcaat tgataataac gaaaatgtct tttaatgatc
     5341 tgggtataat gaggaatttt ccgaacgttt ttactttata tatatatata catgtaacat
     5401 atattctata cgctatagag aaaggaaatt tttcaattaa aaaaaaaata gagaaagagt
     5461 ttcacttctt gattatcgct aacactaatg gttgaagtac tgctacttta attttatgct
     5521 gccaatgaga cgacggggtc atcacggctc atcatgcgcc aaacaaatgt gtgcaataca
     5581 cgctcggatg actgcatgat gaccgcactg actggggaca gcagatccac ctaagcctgt
     5641 gagagaagca gacacccgac agatcaaggc agttaactag tgcactgcag tacaaatcgt
     5701 ccccaacaaa agtgggctct caaaattcat cacatttaaa tgcatatagg aagagcaaca
     5761 gttggtttgc atctgatgtt ccttaaagat ttcgacataa tgtgcgaagt agatcccttc
     5821 agcactgtac ttgtttttgc gactagattg taaatcattc tttatttaat ctctttcttt
     5881 aactactgct taaagtataa tttggtccgt agtttaataa ctatactaag cgaacgctct
     5941 ctcgaaattt ttcttaacgc gtccttgtac tgcgtctaac gcttttgcca cttggatttc
     6001 tattatagga aatagtctca cttactgggc gacgaatttt cgcgttttga tgaagcacag
     6061 gaagaatttc tttatgctca gaagtttgac agcaagcaag ttcatcattc gaactagcct
     6121 tattgtttta gttcagtgac agcgaactgc cgtactcgat gctttatttc tcacggtaga
     6181 gcggaagaac agatagatcc ctacatattg ctaccacttc tattacacaa tagtttcaat
     6241 agcttgcagc gtagctaaac tctaaaattt atctaaatca ctcatataaa ccgaaccctt
     6301 ccccttccgc ttatagtaca gctgatgctc cttcactatt ttaacatgtg gaattcttga
     6361 aagaatgaaa tcgccatgcc aagccatcac acggtctttt atgcaattga ttgaccgcct
     6421 gcaacacata ggcagtaaaa tttttacaag cagtagcaga acaggccaca caatcgcaag
     6481 tgattaacgt ccacacaggt atagggtttc tggaccatat gatacatgct ctggccaagc
     6541 attccggctg gtcgctaatc gttgagtgcc gagcggccgc gattatcaaa aaggatcttc
     6601 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     6661 acttggtctg acag
//
