LOCUS       sequence-360833-        6377 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vector containing a yeast integration cassette for RPL18B-driven 
            expression of SNF7-Link-Neon(gfp), a marker for the endosome, class 
            E compartment.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6377)
  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 6377)
  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: pBBK57 pHomL-RPL18Bp-SNF7-Link-Neon(gfp)-SSA1t...
FEATURES             Location/Qualifiers
     source          1..6377
                     /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      4530..4539
                     /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 tggtcatcac
     3541 tttttggttg gacatcaagt aatgccaaga ataaagagtc accaacaaag gccatagtgc
     3601 ggttgaggga gcatatcaac cttctatcca aaaagcaatc gcatttacgt actcaaatta
     3661 caaatcaaga gaatgaagct agaatctttt tgacgaaggg caataaagta atggcgaaga
     3721 atgcacttaa aaagaagaag acaatcgaac aacttttaag taaggtagaa ggcacaatgg
     3781 agtctatgga acagcagcta ttctctatag aaagtgcaaa cttaaatcta gagacaatga
     3841 gggctatgca ggaaggtgct aaggcaatga aaactattca cagtggcctt gacatagata
     3901 aagtggatga aactatggac gagataaggg agcaagtcga attaggagat gaaataagcg
     3961 acgctatatc caggccttta attactgggg caaacgaggt ggatgaagat gagctggacg
     4021 aggaattgga catgctggct caagaaaatg ctaaccaaga aacgtccaag atcgttaata
     4081 ataatgttaa tgcggcgcct atctcagaga acaaagtctc actacctagt gttccaagta
     4141 ataaaattaa acaaagtgag aactctgtga aggacgggga agaggaagag gatgaagaag
     4201 atgaagatga aaaagcatta agagaactac aagcagaaat ggggcttgga tccggtcctt
     4261 ctggcccaac tggcatggtt tctaagggtg aagaagataa catggcttct ttgccagcca
     4321 cccacgagtt gcacatcttc ggttccatta acggtgtcga cttcgatatg gtcggtcaag
     4381 gtaccggtaa cccaaacgac ggttacgaag aattgaactt gaagtctacc aagggcgact
     4441 tgcagttctc cccttggatt ttggttccac acatcggtta cggtttccac caatacttgc
     4501 catacccaga cggtatgtcc ccattccaag ccgccatggt tgacggttct ggttaccaag
     4561 tccacagaac catgcaattc gaagatggtg cttccttgac cgtgaactac agatacacct
     4621 acgaaggttc tcacatcaag ggcgaagccc aagttaaggg taccggtttc cctgctgacg
     4681 gtccagtcat gactaactcc ttgaccgctg ctgactggtg tcgttccaag aaaacctacc
     4741 caaacgataa gaccatcatt tccactttca agtggtccta caccaccggt aacggcaaga
     4801 gataccgctc taccgctcgt accacctaca ccttcgccaa gccaatggct gccaactact
     4861 tgaagaacca gcctatgtac gttttccgta agaccgagct gaagcattcc aagactgaat
     4921 tgaacttcaa ggaatggcaa aaggctttca ccgacgtcat gggtatggac gaattgtaca
     4981 agtaactcga gtggcgccaa ttggtgcggc aattgataat aacgaaaatg tcttttaatg
     5041 atctgggtat aatgaggaat tttccgaacg tttttacttt atatatatat atacatgtaa
     5101 catatattct atacgctata gagaaaggaa atttttcaat taaaaaaaaa atagagaaag
     5161 agtttcactt cttgattatc gctaacacta atggttgaag tactgctact ttaattttat
     5221 gctgccaatg agacgacggg gtcatcacgg ctcatcatgc gccaaacaaa tgtgtgcaat
     5281 acacgctcgg atgactgcat gatgaccgca ctgactgggg acagcagatc cacctaagcc
     5341 tgtgagagaa gcagacaccc gacagatcaa ggcagttaac tagtgcactg cagtacaaat
     5401 cgtccccaac aaaagtgggc tctcaaaatt catcacattt aaatgcatat aggaagagca
     5461 acagttggtt tgcatctgat gttccttaaa gatttcgaca taatgtgcga agtagatccc
     5521 ttcagcactg tacttgtttt tgcgactaga ttgtaaatca ttctttattt aatctctttc
     5581 tttaactact gcttaaagta taatttggtc cgtagtttaa taactatact aagcgaacgc
     5641 tctctcgaaa tttttcttaa cgcgtccttg tactgcgtct aacgcttttg ccacttggat
     5701 ttctattata ggaaatagtc tcacttactg ggcgacgaat tttcgcgttt tgatgaagca
     5761 caggaagaat ttctttatgc tcagaagttt gacagcaagc aagttcatca ttcgaactag
     5821 ccttattgtt ttagttcagt gacagcgaac tgccgtactc gatgctttat ttctcacggt
     5881 agagcggaag aacagataga tccctacata ttgctaccac ttctattaca caatagtttc
     5941 aatagcttgc agcgtagcta aactctaaaa tttatctaaa tcactcatat aaaccgaacc
     6001 cttccccttc cgcttatagt acagctgatg ctccttcact attttaacat gtggaattct
     6061 tgaaagaatg aaatcgccat gccaagccat cacacggtct tttatgcaat tgattgaccg
     6121 cctgcaacac ataggcagta aaatttttac aagcagtagc agaacaggcc acacaatcgc
     6181 aagtgattaa cgtccacaca ggtatagggt ttctggacca tatgatacat gctctggcca
     6241 agcattccgg ctggtcgcta atcgttgagt gccgagcggc cgcgattatc aaaaaggatc
     6301 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     6361 taaacttggt ctgacag
//
