LOCUS       sequence-270187-        4865 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tagging genes in Cterm with mScarletI in fission yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4865)
  TITLE     pFA6a-mScarletI-KanMX6
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4865)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pFA6a-mScarletI-KanMX6
FEATURES             Location/Qualifiers
     source          1..4865
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     100..119
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        200..218
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     200..217
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             279..971
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet-I
                     /note="matures faster than mScarlet"
                     /translation="VSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYYKQSFPEGFKWERVMNFEDGGA
                     VTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIKM
                     ALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRHS
                     TGGMDELYK"
     primer_bind     complement(409..427)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     677..696
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     terminator      995..1182
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1050..1074)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            1257..2614
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        1257..1600
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1601..2410
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1668..1687)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2416..2614
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2718..2737)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2719..2737)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2824..2841)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2995..3583)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3075..3094)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3754..4545,4546..4614))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGSRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4377..4396
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4615..4719)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4787..4805
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4843..4865)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
       61 cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac
      121 tgagagtgca ccatatggac atattgtcgt tagaacgcgg ctacaattaa tacataacct
      181 tatgtatcat acacatacga tttaggtgac actatagaac gcggccgcca gctgaagctt
      241 cgtacgctgc aggtcgacgg atccccgggt taattaacgt gagcaagggc gaggcagtga
      301 tcaaggagtt catgcggttc aaggtgcaca tggagggctc catgaacggc cacgagttcg
      361 agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg
      421 tgaccaaggg tggccccctg cccttctcct gggacatcct gtcccctcag ttcatgtacg
      481 gctccagggc cttcatcaag caccccgccg acatccccga ctactataag cagtccttcc
      541 ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggcgcc gtgaccgtga
      601 cccaggacac ctccctggag gacggcaccc tgatctacaa ggtgaagctc cgcggcacca
      661 acttccctcc tgacggcccc gtaatgcaga agaagacaat gggctgggaa gcgtccaccg
      721 agcggttgta ccccgaggac ggcgtgctga agggcgacat taagatggcc ctgcgcctga
      781 aggacggcgg ccgctacctg gcggacttca agaccaccta caaggccaag aagcccgtgc
      841 agatgcccgg cgcctacaac gtcgaccgca agttggacat cacctcccac aacgaggact
      901 acaccgtggt ggaacagtac gaacgctccg agggccgcca ctccaccggc ggcatggacg
      961 agctgtacaa gtagggcgcg ccacttctaa ataagcgaat ttcttatgat ttatgatttt
     1021 tattattaaa taagttataa aaaaaataag tgtatacaaa ttttaaagtg actcttaggt
     1081 tttaaaacga aaattcttat tcttgagtaa ctctttcctg taggtcaggt tgctttctca
     1141 ggtatagtat taggtcgctc ttattgacca cacctctacc ggcagatccg ctagggataa
     1201 cagggtaata tagatctgtt tagcttgcct cgtccccgcc gggtcacccg gccagcgaca
     1261 tggaggccca gaataccctc cttgacagtc ttgacgtgcg cagctcaggg gcatgatgtg
     1321 actgtcgccc gtacatttag cccatacatc cccatgtata atcatttgca tccatacatt
     1381 ttgatggccg cacggcgcga agcaaaaatt acggctcctc gctgcagacc tgctagcagg
     1441 gaaacgctcc cctcacagac gcgttgaatt gtccccacgc cgcgcccctg tagagaaata
     1501 taaaaggtta ggatttgcca ctgaggttct tctttcatat acttcctttt aaaatcttgc
     1561 taggatacag ttctcacatc acatccgaac ataaacaacc atgggtaagg aaaagactca
     1621 cgtttcgagg ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc
     1681 tcgcgataat gtcgggcaat caggtgcgac aatctatcga ttgtatggga agcccgatgc
     1741 gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat
     1801 ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc attttatccg
     1861 tactcctgat gatgcatggt tactcaccac tgcgatcccc ggcaaaacag cattccaggt
     1921 attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg
     1981 ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct
     2041 cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga
     2101 gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataagcttt tgccattctc
     2161 accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg
     2221 gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct
     2281 tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca
     2341 aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga
     2401 gtttttctaa tcagtactga caataaaaag attcttgttt tcaagaactt gtcatttgta
     2461 tagttttttt atattgtagt tgttctattt taatcaaatg ttagcgtgat ttatattttt
     2521 ttttcgcctc gacatcatct gcccagatgc gaagttaagt gcgcagaaag taatatcatg
     2581 cgtcaatcgt atgtgaatgc tggtcgctat actgctgtcg attcgatact aacgccgcca
     2641 tccagtttaa acgagctcga attcatcgat gatatcagat ccactagtgg cctatgcggc
     2701 cgcggatctg ccggtctccc tatagtgagt cgtattaatt tcgataagcc aggttaacct
     2761 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     2821 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     2881 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     2941 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     3001 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     3061 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     3121 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     3181 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     3241 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     3301 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     3361 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     3421 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     3481 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     3541 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     3601 ttctacgggg tcttacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     3661 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     3721 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     3781 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     3841 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     3901 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     3961 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     4021 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     4081 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     4141 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     4201 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     4261 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     4321 attctgagaa tagtgtatgc ggctaccgag ttgctcttgc ccggcgtcaa tacgggataa
     4381 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     4441 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     4501 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     4561 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     4621 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     4681 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     4741 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     4801 gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
     4861 cccgg
//
