LOCUS       sequence-455093-        6072 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expressing mTagBFP2-FrFAST under the control of the TEF1 promtoer in
            Saccharomyces cerevisiae.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6072)
  AUTHORS   Evans S, Tominaga M, Lu Z, Bandari NC, McDonnell L, Wang C, Speight 
            RE, Ishii J, Vickers CE, Peng B
  TITLE     Characterization of Fluorescent Reporters for Flow Cytometry-Based 
            Single-Cell Studies in Saccharomyces cerevisiae.
  JOURNAL   ACS Synth Biol. 2025 Sep 19;14(9):3804-3812. doi: 
            10.1021/acssynbio.5c00432. Epub 2025 Sep 10.
  PUBMED    40927980
REFERENCE   2  (bases 1 to 6072)
  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: 
            "10.1021/acssynbio.5c00432"; journalName: "ACS Synth Biol"; date: 
            "2025-09-19- 19"; volume: "14"; issue: "9"; pages: "3804-3812"
COMMENT     Sequence Label: pILFPmTagBFP2FrFAST
FEATURES             Location/Qualifiers
     source          1..6072
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      425..695
                     /gene="S. cerevisae PGK1"
                     /label=PGK1 terminator
                     /note="transcription terminator for 3-phosphoglycerate 
                     kinase gene"
     primer_bind     complement(960..976)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(960..976)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(973..995)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    984..1000
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(1008..1014,1015..1032,1033..1038))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1053..1074
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1191..1208)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1362..1950)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1442..1461)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2121..2912,2913..2981))
                     /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     2744..2763
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2982..3086)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3154..3172
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3210..3232)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3332..3351
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3545..3567
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3559..3576
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3560..3576
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4690..4723)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        4943..5356
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
ORIGIN
        1 catcaaagtt gggtcacaag ttgaacggtg gtatggaaca tgttgctttc ggttccgaag
       61 atattgaaaa cactttggcc aagatggatg atggtcaatt agatggtttg gcttttggtg
      121 ctattcaatt agacggtgac ggtaatatct tgcagtataa tgctgctgaa ggtgacatta
      181 ctggtagaga tccaaaacaa gttatcggca agaacttgtt caaagatgtt gctccaggta
      241 ctgtttcttc tggtttctat ggcaagttca aagaaggtgt tgcttctggt aatctgaaca
      301 ctatgttcga atggatgatt ccaacttcaa gaggtccaac taaggttaag attcacatga
      361 agaaggcttt gtctggcgat tcttattggg ttttcgttaa gagagtctaa agatctgatc
      421 ttaaattgaa ttgaattgaa atcgatagat caattttttt cttttctctt tccccatcct
      481 ttacgctaaa ataatagttt attttatttt ttgaatattt tttatttata tacgtatata
      541 tagactatta tttatctttt aatgattatt aagattttta ttaaaaaaaa attcgctcct
      601 cttttaatgc ctttatgcag tttttttttc ccattcgata tttctatgtt cgggttcagc
      661 gtattttaag tttaataact cgaaaattct gcgttcgtta aagctttcga gaaggatatt
      721 atttactagt aggactattt gcaaagggaa gggatgctaa ggtagagggt gaacgttaca
      781 gaaaagcagg ctgggaagca tatttgagaa gatgcggcca gcaaaactaa aaaactgtat
      841 tataagtaaa tgcatgtata ctaaactcac aaattagagc ttcaatttaa ttatatcagt
      901 tattacccgg gaatctcggt cgtaatgatt taaatgcatg caagcttggc gtaatcatgg
      961 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     1021 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     1081 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     1141 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     1201 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     1261 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     1321 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     1381 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     1441 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     1501 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     1561 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     1621 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     1681 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     1741 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     1801 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     1861 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     1921 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     1981 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     2041 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     2101 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     2161 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     2221 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     2281 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     2341 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     2401 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     2461 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     2521 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     2581 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     2641 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     2701 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     2761 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     2821 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     2881 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     2941 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     3001 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     3061 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
     3121 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
     3181 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
     3241 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
     3301 gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg
     3361 caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc
     3421 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta
     3481 ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg
     3541 ttttcccagt cacgacgttg taaaacgacg gccagtgaat tcatttaaat ttttttgatt
     3601 cggtaatctc cgaacagaag gaagaacgaa ggaaggagca cagacttaga ttggtatata
     3661 tacgcatatg tagtgttgaa gaaacatgaa attgcccagt attcttaacc caactgcaca
     3721 gaacaaaaac ctgcaggaaa cgaagataaa acatgtccac aaaatcatat accagtagag
     3781 ctgagactca tgcaagtccg gttgcatcga aacttttacg tttaatggat gaaaagaaga
     3841 ccaatttgtg tgcttctctt gacgttcgtt cgactgatga gctattgaaa cttgttgaaa
     3901 cgttgggtcc atacatttgc cttttgaaaa cacacgttga tatcttggat gatttcagtt
     3961 atgagggtac tgtcgttcca ttgaaagcat tggcagagaa atacaagttc ttgatatttg
     4021 aggacagaaa attcgccgat atcggtaaca cagtcaaatt acaatataca tcgggcgttt
     4081 accgtatcgc agaatggtct gatatcacca acgcccacgg ggttactggt gctggtattg
     4141 ttgctggctt gaaacaaggt gcgcaagagg tcaccaaaga accaagggga ttattgatgc
     4201 ttgctgaatt gtcttccaag ggttctctag cacacggtga atatactaag ggtaccgttg
     4261 atattgcaaa gagtgataaa gatttcgtta ttgggttcat tgctcagaac gatatgggag
     4321 gaagagaaga agggtttgat tggctaatca tgaccccagg tgtaggttta gacgacaaag
     4381 gcgatgcatt gggtcagcag tacagaaccg tcgacgaagt tgtaagtggt ggatcagata
     4441 tcatcattgt tggcagagga cttttcgcca agggtagaga tcctaaggtt gaaggtgaaa
     4501 gatacagaaa tgctggatgg gaagcgtacc aaaagagaat cagcgctccc cattaattat
     4561 acaggaaact taatagaaca aatcacatat ttaatctaat agccacctgc attggcacgg
     4621 tgcaacacta cttcaacttc atcttacaaa aagatcacgt gatctgttgt attgggatct
     4681 ctagacctaa taacttcgta tagcatacat tatacgaagt tatattaagg gttgctcgag
     4741 aaagagctcc agaaagcgac cacccaactt tggctgataa tagcgtataa acaatgcata
     4801 ctttgtacgt tcaaaataca atgcagtaga tatatttatg catattacat ataatacata
     4861 tcacatagga agcaacaggc gcgttggact tttaattttc gaggaccgcg aatccttaca
     4921 tcacacccaa tcccccacaa gtgatccccc acacaccata gcttcaaaat gtttctactc
     4981 cttttttact cttccagatt ttctcggact ccgcgcatcg ccgtaccact tcaaaacacc
     5041 caagcacagc atactaaatt tcccctcttt cttcctctag ggtgtcgtta attacccgta
     5101 ctaaaggttt ggaaaagaaa aaagagaccg cctcgtttct ttttcttcgt cgaaaaaggc
     5161 aataaaaatt tttatcacgt ttctttttct tgaaaatttt tttttttgat ttttttctct
     5221 ttcgatgacc tcccattgat atttaagtta ataaacggtc ttcaatttct caagtttcag
     5281 tttcattttt cttgttctat tacaactttt tttacttctt gctcattaga aagaaagcat
     5341 agcaatctaa tctaagtttt aattacaaac tcgagaaagg atccaaaatg gtgtctaagg
     5401 gtgaagagct gatcaaagaa aacatgcaca tgaagttgta catggaaggt actgttgata
     5461 accatcattt caagtgtacc tctgaaggtg aaggtaaacc atacgaaggt actcaaacta
     5521 tgagaatcaa ggttgttgaa ggtggtccat tgccatttgc ttttgatatt ttggctacct
     5581 ccttcttgta cggttctaag actttcatca accacactca aggtatccca gatttcttca
     5641 agcaatcttt tccagaaggt ttcacttggg aaagagttac tacttacgaa gatggtggtg
     5701 ttttgactgc tactcaagat acttctttac aagacggttg cttgatctac aacgttaaga
     5761 ttagaggtgt caacttcact tctaacggtc cagttatgca aaagaaaact ttaggttggg
     5821 aagccttcac cgaaacatta tatccagctg atggtggttt ggaaggtaga aatgatatgg
     5881 ctttgaaatt ggttggtggc tcccatttga ttgctaacgc taagactacc tacaggtcta
     5941 agaagccagc taagaatttg aaaatgccag gtgtttacta cgtggactat agattggaaa
     6001 gaatcaaaga agccaacaac gaaacctacg ttgaacaaca cgaagttgct gttgctagat
     6061 attgtgattt gc
//
