LOCUS       sequence-401343-        4969 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For C-terminal tagging using yeast optimized mRuby3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4969)
  AUTHORS   Seitz BC, Mucelli X, Majano M, Wallis Z, Dodge AC, Carmona C, Durant
            M, Maynard S, Huang LS
  TITLE     Meiosis II spindle disassembly requires two distinct pathways.
  JOURNAL   Mol Biol Cell. 2023 Sep 1;34(10):ar98. doi: 10.1091/mbc.E23-03-0096.
            Epub 2023 Jul 12.
  PUBMED    37436806
REFERENCE   2  (bases 1 to 4969)
  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: "Mol Biol 
            Cell."; date: "2023-09-1"; volume: "34(10):ar98. doi"; pages: " 
            10.1091/mbc.E23-03-0096. Epub 2023 Jul 12"
COMMENT     Sequence Label: pFA6a-yomRuby3-SpHIS5
FEATURES             Location/Qualifiers
     source          1..4969
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     847..866
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        847..865
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     gene            complement(970..2170)
                     /label=HIS3MX6
                     /note="yeast selectable marker encoding the S. pombe his5 
                     gene, which corresponds to S. cerevisiae HIS3"
     terminator      complement(970..1167)
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             complement(1173..1826)
                     /codon_start=1
                     /gene="Schizosaccharomyces pombe his5"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=S. pombe his5
                     /note="yeast auxotrophic marker; corresponds to S. 
                     cerevisiae HIS3"
                     /translation="MGRRAFVERNTNETKISVAIALDKAPLPEESNFIDELITSKHTNQ
                     KGEQVIQVDTGIGFLDHMYHALAKHAGWSLRLYSRGDLIIDDHHTAEDTAIALGIAFKQ
                     AMSNFAGVKRFGHAYCPLDEALSRSVVDLSGRPYAVIDLGLKREKVGELSCEMIPHLLY
                     SFSVAAGITLHVTCLYGSNDHHRAESAFKSLAVAMRAATSLTGSSEVPSTKGVL"
     promoter        complement(1827..2170)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     promoter        complement(3470..3488)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3471..3488)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3569..3588)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3688..3710
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3748..3766)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3834..3938
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3939..4007,4008..4799)
                     /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     complement(4157..4176)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcaggttaa cctggcttat
      841 cgaaattaat acgactcact atagggagac cggcagatcc gcggccgcat aggccactag
      901 tggatctgat atcatcgatg aattcgagct cgtttaaact ggatggcggc gttagtatcg
      961 aatcgacagc agtatagcga ccagcattca catacgattg acgcatgata ttactttctg
     1021 cgcacttaac ttcgcatctg ggcagatgat gtcgaggcga aaaaaaatat aaatcacgct
     1081 aacatttgat taaaatagaa caactacaat ataaaaaaac tatacaaatg acaagttctt
     1141 gaaaacaaga atctttttat tgtcagtact ctttacaaca ctcccttcgt gcttgggact
     1201 tcagaacttc cagtaagact agtagccgcg cgcatggcaa cagccagaga tttaaaagcg
     1261 ctttcagcac gatgatggtc attactacca tataagcagg taacatgcaa agtaattcca
     1321 gctgctaccg aaaaggaata tagtaagtga gggatcattt cacaggacaa ttccccaacc
     1381 ttttcacgct ttaatcccaa atcgataaca gcatagggcc gtcccgacaa gtcaactacg
     1441 cttctagaaa gagcttcgtc aagtggacaa taagcatgtc caaatctttt aacgccggca
     1501 aagttactca tagcctgctt gaatgcaata ccaagtgcaa tagcagtatc ttctgcagtg
     1561 tgatgatcat cgatgattaa atcacctctt gagtaaagtc gtaagctcca gcctgcatgt
     1621 ttagccagtg catgatacat gtgatccaag aatccaattc ccgtgtctac ttggattact
     1681 tgttctccct tttggtttgt atgcttggaa gttataagtt catcaataaa attcgactct
     1741 tcaggtaagg gagctttgtc caaagcgatg gcaacgctga ttttcgtttc gttcgtattt
     1801 ctttctacaa aagccctcct acccatggtt gtttatgttc ggatgtgatg tgagaactgt
     1861 atcctagcaa gattttaaaa ggaagtatat gaaagaagaa cctcagtggc aaatcctaac
     1921 cttttatatt tctctacagg ggcgcggcgt ggggacaatt caacgcgtct gtgaggggag
     1981 cgtttccctg ctcgcaggtc tgcagcgagg agccgtaatt tttgcttcgc gccgtgcggc
     2041 catcaaaatg tatggatgca aatgattata catggggatg tatgggctaa atgtacgggc
     2101 gacagtcaca tcatgcccct gagctgcgca cgtcaagact gtcaaggagg gtattctggg
     2161 cctccatgtc gctggccggg tgacccggcg gggacgaggc aagctaaaca gatctatatt
     2221 accctgttat ccctagcgga tctgtccgca acgcttttct gaacgcccgc cttcgcctct
     2281 cattatcatt ctcatcccaa aagaactaac tgtgcatgtt atttgcaata cttcatatac
     2341 gctctgtatt attaatagta tcattaatta cgtcaattga aattcaaaat atcatctttg
     2401 acagcaacat ctatcctctt agacaactag ggccattgca gtgtctcgaa accattaata
     2461 tcactgaaaa gatgaaaaga aaggcaaata tatattgatc actaattttc taagctaaag
     2521 aatctattcc ccctctgtta aatggaattg tttgaaataa aatattataa aatcagaact
     2581 ttggggggga aacataaaaa aatgagaaaa agaaaacgaa ctaactaatg tttaagtaaa
     2641 agaacaaaaa ggtagaccaa tgtagcgctc ttactttagg cgcgccttac ttatacaatt
     2701 catccatacc accgcctaag ttagaatatt tggctacggc cacctctctc tgtacaacat
     2761 aggtttcgtt atcgctctct tcaattcttt ctagtctgtg gtctacagca tgcactcctg
     2821 gcattttgat attgccaaca gtctttttag aacgataagt ggtaacaaaa ttacaatgca
     2881 aatgacctcc accatcaact ttaagcgcaa tatctgtgta gcctctcaaa cctccatcag
     2941 ctgggtacat catttcagta tttggttccc aacctttggt ctttttctgc atcacaggtc
     3001 cgttagaagg aaaattcacc cctcttactt tcacattata gacaagttcg ccatcttcta
     3061 atgaagtatc ttgggtaact gtgacgacac ccccatcttc gtatcttgta accctttccc
     3121 atgtaaaacc ctctggaaaa gattgcttga aaaagtctgg tatatccgca gggtacttga
     3181 tgaaagttcg tgatccgtac atgaatgacg ttgccagaat gtcaaatgca aatggtagtg
     3241 gtccgccttc aattacttta attctcatag tttgcacacc ttcgtaagga cgaccttcac
     3301 cttcccctgt acacttgaat tggtgaccat taacggagcc ctccatgaca actttcattc
     3361 tcatgttttc cttgattaac tcctctcctt tggacaccat gttaattaaa ccagcaccgt
     3421 caccgatccg tcgacctgca gcgtacgaag cttcagctgg cggccgcgtt ctatagtgtc
     3481 acctaaatcg tatgtgtatg atacataagg ttatgtatta attgtagccg cgttctaacg
     3541 acaatatgtc catatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     3601 agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     3661 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     3721 catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg
     3781 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     3841 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     3901 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     3961 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     4021 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     4081 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     4141 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     4201 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     4261 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     4321 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     4381 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     4441 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     4501 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     4561 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     4621 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     4681 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     4741 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     4801 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     4861 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     4921 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttc
//
