LOCUS       sequence-341919-        5614 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Contains mCherry-sfGFP tandem fluorescent protein timer.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5614)
  AUTHORS   Khmelinskii A, Keller PJ, Bartosik A, Meurer M, Barry JD, Mardin BR,
            Kaufmann A, Trautmann S, Wachsmuth M, Pereira G, Huber W, Schiebel 
            E, Knop M
  TITLE     Tandem fluorescent protein timers for in vivo analysis of protein 
            dynamics.
  JOURNAL   Nat Biotechnol. 2012 Jun 24;30(7):708-14. doi: 10.1038/nbt.2281.
  PUBMED    22729030
REFERENCE   2  (bases 1 to 5614)
  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"; doi: "10.1038/nbt"; 
            journalName: "Nat Biotechnol"; date: "2012-06-24- 24"; volume: "30";
            issue: "7"; pages: "708-14"
COMMENT     Sequence Label: pFA6a-mCherry-sfGFP-kanMX (pMaM17)
FEATURES             Location/Qualifiers
     source          1..5614
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     2..21
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        102..120
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     102..119
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             161..868
                     /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(306..324)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     574..593
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     terminator      1647..1834
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1702..1726)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            1909..3265
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        1909..2252
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             2253..3062
                     /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(2320..2339)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      3068..3265
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(3369..3388)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3370..3388)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3475..3492)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3646..4234)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3726..3745)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4405..5196,5197..5265))
                     /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     5028..5047
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5266..5370)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5438..5456
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5494..5516)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gcggcatcag agcagattgt actgagagtg caccatatgg acatattgtc gttagaacgc
       61 ggctacaatt aatacataac cttatgtatc atacacatac gatttaggtg acactataga
      121 acgcggccgc cagctgaagc ttcgtacgct gcaggtcgac atggtgagca agggcgagga
      181 ggataacatg gccatcatca aggagttcat gcgcttcaag gtgcacatgg agggctccgt
      241 gaacggccac gagttcgaga tcgagggcga gggcgagggc cgcccctacg agggcaccca
      301 gaccgccaag ctgaaggtga ccaagggtgg ccccctgccc ttcgcctggg acatcctgtc
      361 ccctcagttc atgtacggct ccaaggccta cgtgaagcac cccgccgaca tccccgacta
      421 cttgaagctg tccttccccg agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg
      481 cggcgtggtg accgtgaccc aggactcctc cctgcaggac ggcgagttca tctacaaggt
      541 gaagctgcgc ggcaccaact tcccctccga cggccccgta atgcagaaga agaccatggg
      601 ctgggaggcc tcctccgagc ggatgtaccc cgaggacggc gccctgaagg gcgagatcaa
      661 gcagaggctg aagctgaagg acggcggcca ctacgacgct gaggtcaaga ccacctacaa
      721 ggccaagaag cccgtgcagc tgcccggcgc ctacaacgtc aacatcaagt tggacatcac
      781 ctcccacaac gaggactaca ccatcgtgga acagtacgaa cgcgccgagg gccgccactc
      841 caccggcggc atggacgagc tgtacaagct cgacggagca ggtgctggtg ctggtgctgg
      901 agcaatgtcc aagggtgaag agctatttac tggggttgta cccattttgg tagaactgga
      961 cggagatgta aacggacata aattctctgt tagaggtgag ggcgaaggcg atgccaccaa
     1021 tggtaaattg actctgaagt ttatatgcac tacgggtaaa ttacctgttc cttggccaac
     1081 cctagtaaca actttgacat atggtgttca atgtttctca agatacccag accatatgaa
     1141 aaggcatgat ttctttaaaa gtgctatgcc agaaggctac gtgcaagaga gaactatctc
     1201 ctttaaggat gacggtacgt ataaaacacg agcagaagtg aaattcgaag gggatacact
     1261 agttaatcgc atcgaattaa agggtataga ctttaaggaa gatggtaata ttctcggcca
     1321 taaacttgag tataatttca actcgcataa tgtgtacatt acagctgaca aacaaaagaa
     1381 cggaattaaa gcgaatttta aaatcaggca caacgtcgaa gatgggtctg ttcaacttgc
     1441 cgatcattat cagcaaaaca cccctattgg tgatggtcca gtcttgttac ccgataatca
     1501 ctacttaagc acacagtcta gattgtcaaa agatccgaat gaaaagcgtg atcacatggt
     1561 tttattggaa tttgtcaccg ctgcaggaat aactcacgga atggacgagc tttataaggg
     1621 atcctaagcg cgccacttct aaataagcga atttcttatg atttatgatt tttattatta
     1681 aataagttat aaaaaaaata agtgtataca aattttaaag tgactcttag gttttaaaac
     1741 gaaaattctt attcttgagt aactctttcc tgtaggtcag gttgctttct caggtatagt
     1801 atgaggtcgc tcttattgac cacacctcta ccggcagatc cgctagggat aacagggtaa
     1861 tatagatctg tttagcttgc ctcgtccccg ccgggtcacc cggccagcga catggaggcc
     1921 cagaataccc tccttgacag tcttgacgtg cgcagctcag gggcatgatg tgactgtcgc
     1981 ccgtacattt agcccataca tccccatgta taatcatttg catccataca ttttgatggc
     2041 cgcacggcgc gaagcaaaaa ttacggctcc tcgctgcaga cctgcgagca gggaaacgct
     2101 cccctcacag acgcgttgaa ttgtccccac gccgcgcccc tgtagagaaa tataaaaggt
     2161 taggatttgc cactgaggtt cttctttcat atacttcctt ttaaaatctt gctaggatac
     2221 agttctcaca tcacatccga acataaacaa ccatgggtaa ggaaaagact cacgtttcga
     2281 ggccgcgatt aaattccaac atggatgctg atttatatgg gtataaatgg gctcgcgata
     2341 atgtcgggca atcaggtgcg acaatctatc gattgtatgg gaagcccgat gcgccagagt
     2401 tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt tacagatgag atggtcagac
     2461 taaactggct gacggaattt atgcctcttc cgaccatcaa gcattttatc cgtactcctg
     2521 atgatgcatg gttactcacc actgcgatcc ccggcaaaac agcattccag gtattagaag
     2581 aatatcctga ttcaggtgaa aatattgttg atgcgctggc agtgttcctg cgccggttgc
     2641 attcgattcc tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg
     2701 cgcaatcacg aatgaataac ggtttggttg atgcgagtga ttttgatgac gagcgtaatg
     2761 gctggcctgt tgaacaagtc tggaaagaaa tgcataagct tttgccattc tcaccggatt
     2821 cagtcgtcac tcatggtgat ttctcacttg ataaccttat ttttgacgag gggaaattaa
     2881 taggttgtat tgatgttgga cgagtcggaa tcgcagaccg ataccaggat cttgccatcc
     2941 tatggaactg cctcggtgag ttttctcctt cattacagaa acggcttttt caaaaatatg
     3001 gtattgataa tcctgatatg aataaattgc agtttcattt gatgctcgat gagtttttct
     3061 aatcagtact gacaataaaa agattcttgt tttcaagaac ttgtcatttg tatagttttt
     3121 ttatattgta gttgttctat tttaatcaaa tgttagcgtg atttatattt tttttcgcct
     3181 cgacatcatc tgcccagatg cgaagttaag tgcgcagaaa gtaatatcat gcgtcaatcg
     3241 tatgtgaatg ctggtcgcta tactgctgtc gattcgatac taacgccgcc atccagttta
     3301 aacgagctcg aattcatcga tgatatcaga tccactagtg gcctatgcgg ccgcggatct
     3361 gccggtctcc ctatagtgag tcgtattaat ttcgataagc caggttaacc tgcattaatg
     3421 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     3481 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     3541 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     3601 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     3661 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     3721 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     3781 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     3841 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     3901 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     3961 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     4021 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     4081 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     4141 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     4201 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     4261 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     4321 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     4381 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     4441 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     4501 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     4561 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     4621 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     4681 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     4741 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     4801 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     4861 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     4921 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     4981 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     5041 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     5101 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     5161 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     5221 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     5281 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     5341 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     5401 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt
     5461 tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac
     5521 ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc
     5581 gggtgttggc gggtgtcggg gctggcttaa ctat
//
