LOCUS       sequence-172999-        4879 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tag S. pombe genes in C terminal with PAmKate.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4879)
  TITLE     pFA6a-mEGFP-KanMX6
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4879)
  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            "Unpublished"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-PAmKate-KanMX6
FEATURES             Location/Qualifiers
     source          1..4879
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             63..773
                     /codon_start=1
                     /product="monomeric photoactivatable red fluorescent 
                     protein (Gunewardene et al., 2011)"
                     /label=PAmKate
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMR
                     IKVVEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFWKQSFPEGFTWERVTTYEDGGV
                     LTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRGDM
                     ALKLVGGGHLICNLKTTYRSKKPAKNLKMPGVYYVDRRLERIKEADKETYVEQHEVAVA
                     RYCDLPSKLGHKLN"
     terminator      794..981
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(849..873)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            1056..2412
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        1056..1399
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1400..2209
                     /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(1467..1486)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2215..2412
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2516..2535)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2517..2535)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2622..2639)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2793..3381)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2873..2892)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3552..4343,4344..4412))
                     /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     4175..4194
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4413..4517)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4585..4603
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4641..4663)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4763..4782
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(4863..4879,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(4863..4879,1)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg acggatcccc gggttaatta
       61 acgtgtctaa gggcgaagag ctgattaagg agaacatgca catgaagctg tacatggagg
      121 gcaccgtgaa caaccaccac ttcaagtgca catccgaggg cgaaggcaag ccctacgagg
      181 gcacccagac catgagaatc aaggtggtcg agggcggccc tctccccttc gccttcgaca
      241 tcctggctac cagcttcatg tacggcagca agaccttcat caaccacacc cagggcatcc
      301 ccgacttctg gaagcagtcc ttccctgagg gcttcacatg ggagagagtc accacatacg
      361 aagacggggg cgtgctgacc gctacccagg acaccagcct ccaggacggc tgcctcatct
      421 acaacgtcaa gatcagaggg gtgaacttcc catccaacgg ccctgtgatg cagaagaaaa
      481 cactcggctg ggaggccaac accgagatgc tgtaccccgc tgacggcggc ctggaaggca
      541 gaggggacat ggccctgaag ctcgtgggcg ggggccacct gatctgcaac ttgaagacca
      601 catacagatc caagaaaccc gctaagaacc tcaagatgcc cggcgtctac tatgtggacc
      661 gcagactgga aagaatcaag gaggccgaca aagagaccta cgtcgagcag cacgaggtgg
      721 ctgtggccag atactgcgac ctccctagca aactggggca caagcttaat taaggcgcgc
      781 cacttctaaa taagcgaatt tcttatgatt tatgattttt attattaaat aagttataaa
      841 aaaaataagt gtatacaaat tttaaagtga ctcttaggtt ttaaaacgaa aattcttatt
      901 cttgagtaac tctttcctgt aggtcaggtt gctttctcag gtatagtatg aggtcgctct
      961 tattgaccac acctctaccg gcagatccgc tagggataac agggtaatat agatctgttt
     1021 agcttgcctc gtccccgccg ggtcacccgg ccagcgacat ggaggcccag aataccctcc
     1081 ttgacagtct tgacgtgcgc agctcagggg catgatgtga ctgtcgcccg tacatttagc
     1141 ccatacatcc ccatgtataa tcatttgcat ccatacattt tgatggccgc acggcgcgaa
     1201 gcaaaaatta cggctcctcg ctgcagacct gcgagcaggg aaacgctccc ctcacagacg
     1261 cgttgaattg tccccacgcc gcgcccctgt agagaaatat aaaaggttag gatttgccac
     1321 tgaggttctt ctttcatata cttcctttta aaatcttgct aggatacagt tctcacatca
     1381 catccgaaca taaacaacca tgggtaagga aaagactcac gtttcgaggc cgcgattaaa
     1441 ttccaacatg gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc
     1501 aggtgcgaca atctatcgat tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca
     1561 tggcaaaggt agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac
     1621 ggaatttatg cctcttccga ccatcaagca ttttatccgt actcctgatg atgcatggtt
     1681 actcaccact gcgatccccg gcaaaacagc attccaggta ttagaagaat atcctgattc
     1741 aggtgaaaat attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt
     1801 ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat
     1861 gaataacggt ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga
     1921 acaagtctgg aaagaaatgc ataagctttt gccattctca ccggattcag tcgtcactca
     1981 tggtgatttc tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga
     2041 tgttggacga gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct
     2101 cggtgagttt tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc
     2161 tgatatgaat aaattgcagt ttcatttgat gctcgatgag tttttctaat cagtactgac
     2221 aataaaaaga ttcttgtttt caagaacttg tcatttgtat agttttttta tattgtagtt
     2281 gttctatttt aatcaaatgt tagcgtgatt tatatttttt ttcgcctcga catcatctgc
     2341 ccagatgcga agttaagtgc gcagaaagta atatcatgcg tcaatcgtat gtgaatgctg
     2401 gtcgctatac tgctgtcgat tcgatactaa cgccgccatc cagtttaaac gagctcgaat
     2461 tcatcgatga tatcagatcc actagtggcc tatgcggccg cggatctgcc ggtctcccta
     2521 tagtgagtcg tattaatttc gataagccag gttaacctgc attaatgaat cggccaacgc
     2581 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     2641 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     2701 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     2761 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     2821 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     2881 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     2941 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     3001 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     3061 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     3121 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     3181 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     3241 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     3301 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     3361 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     3421 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     3481 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     3541 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     3601 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     3661 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     3721 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     3781 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     3841 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     3901 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     3961 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     4021 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     4081 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     4141 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     4201 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     4261 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     4321 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     4381 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     4441 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     4501 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt
     4561 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt
     4621 ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt
     4681 ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg
     4741 tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg
     4801 gacatattgt cgttagaacg cggctacaat taatacataa ccttatgtat catacacata
     4861 cgatttaggt gacactata
//
