LOCUS       sequence-360863-        6297 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vector containing a yeast integration cassette for RPL18B-driven 
            expression of Scarlet(rfp)-Link-SFT2, a marker for the late Golgi.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6297)
  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 6297)
  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: pBBK74
            pHomL-RPL18Bp-Scarlet(rfp)-Link-SFT2-SS...
FEATURES             Location/Qualifiers
     source          1..6297
                     /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"
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 gttagtaaag
     3541 gtgaagctgt tataaaagaa ttcatgaggt ttaaagttca tatggaaggt tcaatgaatg
     3601 gtcatgaatt tgaaattgaa ggtgaaggtg aaggtagacc atatgaaggt acacaaactg
     3661 ctaaattgaa agttactaaa ggtggtccat taccattttc ttgggatatt ttgtctccac
     3721 aattcatgta tggttctaga gcttttatta agcatccagc tgatattcca gattattata
     3781 aacaatcttt tcctgaaggt tttaaatggg aaagagttat gaatttcgaa gatggtggtg
     3841 ctgttactgt tactcaagat acttctttgg aagatggtac tttaatctat aaagttaaat
     3901 tgagaggtac taattttcca ccagatggtc cagttatgca aaagaaaact atgggttggg
     3961 aagcatctac tgaaagattg tatccagaag atggtgtttt gaaaggagac attaagatgg
     4021 ctttgagatt gaaagatggt ggtcggtacc tggccgactt caagaccacc tataaagcta
     4081 aaaaaccagt tcaaatgcca ggtgcatata atgttgatag aaagttagat ataacgtcgc
     4141 ataacgagga ctatactgta gttgaacaat atgaacgtag tgaaggtaga catagtaccg
     4201 gaggaatgga tgaattgtat aaatctggtc cttctggccc aactggttct atgagcgagg
     4261 aaccaccttc tgaccaggtc aatagtctcc gtgactcatt gaatcgatgg aatcaaacaa
     4321 gacagcagaa ctcgcagggt tttaatgaat ctgcgaagac attgttctca agctgggcgg
     4381 attctctcaa taccagggcc caggatatat atcagacgtt gcctgtatct agacaggact
     4441 tggtgcaaga ccaggagccg tcgtggttcc aattgtcaag aacggaaaga atggtacttt
     4501 ttgtctgttt tcttttgggt gcaacagcct gtttcactct ttgtactttc cttttccccg
     4561 ttctagccgc taaaccaaga aagtttggtt tactatggac aatggggtcc ctactatttg
     4621 ttcttgcgtt tggggtactt atgggaccac tcgcgtactt aaaacatttg actgcaaggg
     4681 aaaggctgcc tttttcgatg ttctttttcg ccacatgctt catgacgatt tatttcgcag
     4741 ccttttccaa gaacacggtg ctgactatta catgtgctct tcttgaatta gttgccgtca
     4801 tttattatgc tatttcatat ttcccattcg gtgcaacagg tttgaggatg ttaagctctg
     4861 ctggtgtcaa ttcggcaaga ggtgttctgc gcatctagcg agatcctaac tcgaggccaa
     4921 ttggtgcggc aattgataat aacgaaaatg tcttttaatg atctgggtat aatgaggaat
     4981 tttccgaacg tttttacttt atatatatat atacatgtaa catatattct atacgctata
     5041 gagaaaggaa atttttcaat taaaaaaaaa atagagaaag agtttcactt cttgattatc
     5101 gctaacacta atggttgaag tactgctact ttaattttat gctgccaatg agacgacggg
     5161 gtcatcacgg ctcatcatgc gccaaacaaa tgtgtgcaat acacgctcgg atgactgcat
     5221 gatgaccgca ctgactgggg acagcagatc cacctaagcc tgtgagagaa gcagacaccc
     5281 gacagatcaa ggcagttaac tagtgcactg cagtacaaat cgtccccaac aaaagtgggc
     5341 tctcaaaatt catcacattt aaatgcatat aggaagagca acagttggtt tgcatctgat
     5401 gttccttaaa gatttcgaca taatgtgcga agtagatccc ttcagcactg tacttgtttt
     5461 tgcgactaga ttgtaaatca ttctttattt aatctctttc tttaactact gcttaaagta
     5521 taatttggtc cgtagtttaa taactatact aagcgaacgc tctctcgaaa tttttcttaa
     5581 cgcgtccttg tactgcgtct aacgcttttg ccacttggat ttctattata ggaaatagtc
     5641 tcacttactg ggcgacgaat tttcgcgttt tgatgaagca caggaagaat ttctttatgc
     5701 tcagaagttt gacagcaagc aagttcatca ttcgaactag ccttattgtt ttagttcagt
     5761 gacagcgaac tgccgtactc gatgctttat ttctcacggt agagcggaag aacagataga
     5821 tccctacata ttgctaccac ttctattaca caatagtttc aatagcttgc agcgtagcta
     5881 aactctaaaa tttatctaaa tcactcatat aaaccgaacc cttccccttc cgcttatagt
     5941 acagctgatg ctccttcact attttaacat gtggaattct tgaaagaatg aaatcgccat
     6001 gccaagccat cacacggtct tttatgcaat tgattgaccg cctgcaacac ataggcagta
     6061 aaatttttac aagcagtagc agaacaggcc acacaatcgc aagtgattaa cgtccacaca
     6121 ggtatagggt ttctggacca tatgatacat gctctggcca agcattccgg ctggtcgcta
     6181 atcgttgagt gccgagcggc cgcgattatc aaaaaggatc ttcacctaga tccttttaaa
     6241 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacag
//
