LOCUS       sequence-149724-        4677 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  alias pFA6a-mCherry-kanMX6 - vector for C-terminal tagging of yeast 
            genes with mCherry and G418 selection marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4677)
  AUTHORS   Malcova I, Farkasovsky M, Senohrabkova L, Vasicova P, Hasek J
  TITLE     New integrative modules for multicolor-protein labeling and 
            live-cell imaging in Saccharomyces cerevisiae.
  JOURNAL   FEMS Yeast Res. 2016 Mar 17. pii: fow027.
  PUBMED    26994102
REFERENCE   2  (bases 1 to 4677)
  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: "FEMS Yeast 
            Res. 2016 Mar 17. pii: fow027."
COMMENT     Sequence Label: pFM571
FEATURES             Location/Qualifiers
     source          1..4677
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             70..780
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(215..233)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     483..502
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     gene            854..2210
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        854..1197
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1198..2007
                     /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(1265..1284)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2013..2210
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2314..2333)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2315..2333)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2420..2437)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2591..3179)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2671..2690)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3350..4141,4142..4210))
                     /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     3973..3992
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4211..4315)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4383..4401
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4439..4461)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4561..4580
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(4661..4677,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(4661..4677,1)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg acggtgcagg tgctggagct
       61 ggtgctggta tggtgagcaa gggcgaggag gataacatgg ccatcatcaa ggagttcatg
      121 cgcttcaagg tgcacatgga gggctccgtg aacggccacg agttcgagat cgagggcgag
      181 ggcgagggcc gcccctacga gggcacccag accgccaagc tgaaggtgac caagggtggc
      241 cccctgccct tcgcctggga catcctgtcc cctcagttca tgtacggctc caaggcctac
      301 gtgaagcacc ccgccgacat ccccgactac ttgaagctgt ccttccccga gggcttcaag
      361 tgggagcgcg tgatgaactt cgaggacggc ggcgtggtga ccgtgaccca ggactcctcc
      421 ctgcaggacg gcgagttcat ctacaaggtg aagctgcgcg gcaccaactt cccctccgac
      481 ggccccgtaa tgcagaagaa gaccatgggc tgggaggcct cctccgagcg gatgtacccc
      541 gaggacggcg ccctgaaggg cgagatcaag cagaggctga agctgaagga cggcggccac
      601 tacgacgctg aggtcaagac cacctacaag gccaagaagc ccgtgcagct gcccggcgcc
      661 tacaacgtca acatcaagtt ggacatcacc tcccacaacg aggactacac catcgtggaa
      721 cagtacgaac gcgccgaggg ccgccactcc accggcggca tggacgagct gtacaagtaa
      781 ggatccgcta gggataacag ggtaatatag atctgtttag cttgcctcgt ccccgccggg
      841 tcacccggcc agcgacatgg aggcccagaa taccctcctt gacagtcttg acgtgcgcag
      901 ctcaggggca tgatgtgact gtcgcccgta catttagccc atacatcccc atgtataatc
      961 atttgcatcc atacattttg atggccgcac ggcgcgaagc aaaaattacg gctcctcgct
     1021 gcagacctgc gagcagggaa acgctcccct cacagacgcg ttgaattgtc cccacgccgc
     1081 gcccctgtag agaaatataa aaggttagga tttgccactg aggttcttct ttcatatact
     1141 tccttttaaa atcttgctag gatacagttc tcacatcaca tccgaacata aacaaccatg
     1201 ggtaaggaaa agactcacgt ttcgaggccg cgattaaatt ccaacatgga tgctgattta
     1261 tatgggtata aatgggctcg cgataatgtc gggcaatcag gtgcgacaat ctatcgattg
     1321 tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag cgttgccaat
     1381 gatgttacag atgagatggt cagactaaac tggctgacgg aatttatgcc tcttccgacc
     1441 atcaagcatt ttatccgtac tcctgatgat gcatggttac tcaccactgc gatccccggc
     1501 aaaacagcat tccaggtatt agaagaatat cctgattcag gtgaaaatat tgttgatgcg
     1561 ctggcagtgt tcctgcgccg gttgcattcg attcctgttt gtaattgtcc ttttaacagc
     1621 gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg
     1681 agtgattttg atgacgagcg taatggctgg cctgttgaac aagtctggaa agaaatgcat
     1741 aagcttttgc cattctcacc ggattcagtc gtcactcatg gtgatttctc acttgataac
     1801 cttatttttg acgaggggaa attaataggt tgtattgatg ttggacgagt cggaatcgca
     1861 gaccgatacc aggatcttgc catcctatgg aactgcctcg gtgagttttc tccttcatta
     1921 cagaaacggc tttttcaaaa atatggtatt gataatcctg atatgaataa attgcagttt
     1981 catttgatgc tcgatgagtt tttctaatca gtactgacaa taaaaagatt cttgttttca
     2041 agaacttgtc atttgtatag tttttttata ttgtagttgt tctattttaa tcaaatgtta
     2101 gcgtgattta tatttttttt cgcctcgaca tcatctgccc agatgcgaag ttaagtgcgc
     2161 agaaagtaat atcatgcgtc aatcgtatgt gaatgctggt cgctatactg ctgtcgattc
     2221 gatactaacg ccgccatcca gtttaaacga gctcgaattc atcgatgata tcagatccac
     2281 tagtggccta tgcggccgcg gatctgccgg tctccctata gtgagtcgta ttaatttcga
     2341 taagccaggt taacctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     2401 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     2461 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     2521 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     2581 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     2641 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     2701 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     2761 cttcgggaag cgtggcgctt tctcaatgct cacgctgtag gtatctcagt tcggtgtagg
     2821 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     2881 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     2941 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     3001 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
     3061 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     3121 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     3181 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     3241 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     3301 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     3361 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     3421 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     3481 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     3541 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     3601 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     3661 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     3721 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     3781 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     3841 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     3901 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     3961 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     4021 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     4081 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     4141 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     4201 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     4261 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     4321 ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata
     4381 aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc
     4441 tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca
     4501 gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg
     4561 cggcatcaga gcagattgta ctgagagtgc accatatgga catattgtcg ttagaacgcg
     4621 gctacaatta atacataacc ttatgtatca tacacatacg atttaggtga cactata
//
