LOCUS       sequence-446736-        5667 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expressing mScarletI3 under the control of the TEF1 promoter in 
            Saccharomyces cerevisiae.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5667)
  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 5667)
  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: pILFPQ13A7
FEATURES             Location/Qualifiers
     source          1..5667
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      9..279
                     /gene="S. cerevisae PGK1"
                     /label=PGK1 terminator
                     /note="transcription terminator for 3-phosphoglycerate 
                     kinase gene"
     primer_bind     complement(544..560)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(544..560)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(557..579)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    568..584
                     /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(592..598,599..616,617..622))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    637..658
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(775..792)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(946..1534)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1026..1045)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1705..2496,2497..2565))
                     /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     2328..2347
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2566..2670)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2738..2756
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2794..2816)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2916..2935
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3129..3151
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3143..3160
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3144..3160
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4274..4307)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        4527..4940
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
ORIGIN
        1 gatcttaaat tgaattgaat tgaaatcgat agatcaattt ttttcttttc tctttcccca
       61 tcctttacgc taaaataata gtttatttta ttttttgaat attttttatt tatatacgta
      121 tatatagact attatttatc ttttaatgat tattaagatt tttattaaaa aaaaattcgc
      181 tcctctttta atgcctttat gcagtttttt tttcccattc gatatttcta tgttcgggtt
      241 cagcgtattt taagtttaat aactcgaaaa ttctgcgttc gttaaagctt tcgagaagga
      301 tattatttac tagtaggact atttgcaaag ggaagggatg ctaaggtaga gggtgaacgt
      361 tacagaaaag caggctggga agcatatttg agaagatgcg gccagcaaaa ctaaaaaact
      421 gtattataag taaatgcatg tatactaaac tcacaaatta gagcttcaat ttaattatat
      481 cagttattac ccgggaatct cggtcgtaat gatttaaatg catgcaagct tggcgtaatc
      541 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
      601 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
      661 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
      721 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
      781 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
      841 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
      901 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
      961 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     1021 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     1081 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     1141 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     1201 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     1261 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     1321 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     1381 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     1441 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     1501 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     1561 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     1621 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     1681 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     1741 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     1801 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     1861 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     1921 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     1981 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     2041 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     2101 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     2161 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     2221 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     2281 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     2341 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     2401 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     2461 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     2521 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     2581 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     2641 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     2701 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt
     2761 tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac
     2821 ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc
     2881 gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag
     2941 agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag
     3001 gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
     3061 gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc
     3121 agggttttcc cagtcacgac gttgtaaaac gacggccagt gaattcattt aaattttttt
     3181 gattcggtaa tctccgaaca gaaggaagaa cgaaggaagg agcacagact tagattggta
     3241 tatatacgca tatgtagtgt tgaagaaaca tgaaattgcc cagtattctt aacccaactg
     3301 cacagaacaa aaacctgcag gaaacgaaga taaaacatgt ccacaaaatc atataccagt
     3361 agagctgaga ctcatgcaag tccggttgca tcgaaacttt tacgtttaat ggatgaaaag
     3421 aagaccaatt tgtgtgcttc tcttgacgtt cgttcgactg atgagctatt gaaacttgtt
     3481 gaaacgttgg gtccatacat ttgccttttg aaaacacacg ttgatatctt ggatgatttc
     3541 agttatgagg gtactgtcgt tccattgaaa gcattggcag agaaatacaa gttcttgata
     3601 tttgaggaca gaaaattcgc cgatatcggt aacacagtca aattacaata tacatcgggc
     3661 gtttaccgta tcgcagaatg gtctgatatc accaacgccc acggggttac tggtgctggt
     3721 attgttgctg gcttgaaaca aggtgcgcaa gaggtcacca aagaaccaag gggattattg
     3781 atgcttgctg aattgtcttc caagggttct ctagcacacg gtgaatatac taagggtacc
     3841 gttgatattg caaagagtga taaagatttc gttattgggt tcattgctca gaacgatatg
     3901 ggaggaagag aagaagggtt tgattggcta atcatgaccc caggtgtagg tttagacgac
     3961 aaaggcgatg cattgggtca gcagtacaga accgtcgacg aagttgtaag tggtggatca
     4021 gatatcatca ttgttggcag aggacttttc gccaagggta gagatcctaa ggttgaaggt
     4081 gaaagataca gaaatgctgg atgggaagcg taccaaaaga gaatcagcgc tccccattaa
     4141 ttatacagga aacttaatag aacaaatcac atatttaatc taatagccac ctgcattggc
     4201 acggtgcaac actacttcaa cttcatctta caaaaagatc acgtgatctg ttgtattggg
     4261 atctctagac ctaataactt cgtatagcat acattatacg aagttatatt aagggttgct
     4321 cgagaaagag ctccagaaag cgaccaccca actttggctg ataatagcgt ataaacaatg
     4381 catactttgt acgttcaaaa tacaatgcag tagatatatt tatgcatatt acatataata
     4441 catatcacat aggaagcaac aggcgcgttg gacttttaat tttcgaggac cgcgaatcct
     4501 tacatcacac ccaatccccc acaagtgatc ccccacacac catagcttca aaatgtttct
     4561 actccttttt tactcttcca gattttctcg gactccgcgc atcgccgtac cacttcaaaa
     4621 cacccaagca cagcatacta aatttcccct ctttcttcct ctagggtgtc gttaattacc
     4681 cgtactaaag gtttggaaaa gaaaaaagag accgcctcgt ttctttttct tcgtcgaaaa
     4741 aggcaataaa aatttttatc acgtttcttt ttcttgaaaa tttttttttt tgattttttt
     4801 ctctttcgat gacctcccat tgatatttaa gttaataaac ggtcttcaat ttctcaagtt
     4861 tcagtttcat ttttcttgtt ctattacaac tttttttact tcttgctcat tagaaagaaa
     4921 gcatagcaat ctaatctaag ttttaattac aaactcgaga aaggatccaa aatggattct
     4981 accgaagctg tcatcaaaga gttcatgaga ttcaaggttc acatggaagg ttctatgaac
     5041 ggtcacgaat ttgaaattga aggtgaaggt gagggtagac catacgaagg tactcaaact
     5101 gcaaaactga aagttacaaa aggtggtcca ttgccattct cttgggatat tttgtctcca
     5161 caattcatgt acggttccag agcttttatc aaacatccag ctgatattcc agactactgg
     5221 aagcaatcat ttcctgaagg ttttaagtgg gagagagtca tgatttttga agatggtggt
     5281 actgtctctg ttacccaaga tacttctttg gaagatggca ctttgatcta caaggttaag
     5341 ttgagaggtg gtaattttcc accagatggt ccagttatgc agaaaagaac tatgggttgg
     5401 gaagcctcta ctgaaagatt atatccagaa gatgttgtct tgaagggcga tattaagatg
     5461 gctttgagat tgaaagacgg tggtagatat ttggctgatt tcaagaccac ttacaaggcc
     5521 aaaaagccag ttcaaatgcc aggtgctttt aacattgata ggaagttgga tattaccagc
     5581 cacaacgaag attacaccgt tgttgaacaa tacgaaagat ccgttgctag acattctact
     5641 ggtggttcag gtggttctta aagatct
//
