LOCUS       sequence-407702-        7616 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  express ymTq2-d11 as donor control for yAT1.03.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7616)
  AUTHORS   Luzia L, Battjes J, Zwering E, Jansen D, Melkonian C, Teusink B
  TITLE     A fast method to distinguish between fermentative and respiratory 
            metabolisms in single yeast cells.
  JOURNAL   iScience. 2023 Dec 21;27(1):108767. doi: 10.1016/j.isci.2023.108767.
            eCollection 2024 Jan 19.
  PUBMED    38235328
REFERENCE   2  (bases 1 to 7616)
  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: "iScience.";
            date: "2023-12-21"; volume: "27(1):108767. doi"; pages: " 
            10.1016/j.isci.2023.108767. eCollection 2024 Jan 19"
COMMENT     Sequence Label: pDRF1-GW ymTq2-d11
FEATURES             Location/Qualifiers
     source          1..7616
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(1506..1530)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(1592..1611)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1593..1611)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      1759..1946
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1814..1838)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(1958..1975)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1958..1974)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1967..1989)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2410..3756
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             complement(3867..4670)
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(4671..4886)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(4911..4929)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4997..5101
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5102..5170,5171..5962)
                     /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     complement(5320..5339)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6133..6721
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6622..6641
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6875..6892
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7009..7030
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7045..7050,7051..7068,7069..7075)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7083..7099
                     /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)."
     primer_bind     7088..7110
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7107..7123
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7107..7123
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     rep_origin      complement(7153..7608)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7290..7311)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7502..7521
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 caagcttcct gaaacggaga aacataaaca ggcattgctg ggatcaccca tacatcactc
       61 tgttttgcct gaccttttcc ggtaatttga aaacaaaccc ggtctcgaag cggagatccg
      121 gcgataatta ccgcagaaat aaacccatac acgagacgta gaaccagccg cacatggccg
      181 gagaaactcc tgcgagaatt tcgtaaactc gcgcgcattg catctgtatt tcctaatgcg
      241 gcacttccag gcctcgatcg agaccgttta tccattgctt ttttgttgtc tttttccctc
      301 gttcacagaa agtctgaaga agctatagta gaactatgag ctttttttgt ttctgttttc
      361 cttttttttt tttttacctc tgtggaaatt gttactctca cactctttag ttcgtttgtt
      421 tgttttgttt attccaatta tgaccggtga cgaaacgtgg tcgatggtgg gtaccgctta
      481 tgctcccctc cattagtttc gattatataa aaaggccaaa tattgtatta ttttcaaatg
      541 tcctatcatt atcgtctaac atctaatttc tcttaaattt tttctctttc tttcctataa
      601 caccaatagt gaaaatcttt ttttcttcta tatctacaaa aacttttttt ttctatcaac
      661 ctcgttgata aattttttct ttaacaatcg ttaataatta attaattgga aaataaccat
      721 tttttctctc ttttatacac acattcaaaa gaaagaaaaa aaatataccc cagcctcgac
      781 tagtgctagc actagtaagc ttttaattaa atggttagta aaggtgaaga attgttcact
      841 ggtgtagtgc caattttagt agaattagat ggtgacgtta acggtcataa gttctctgta
      901 agcggtgaag gtgaaggtga cgcaacatac ggtaaattaa ctttgaaatt catttgtact
      961 acaggtaaat taccagttcc atggccaact ttggttacta ctttgtcttg gggtgttcaa
     1021 tgtttcgcta gatacccaga tcatatgaag caacatgatt tctttaagtc tgctatgcca
     1081 gaaggttacg ttcaagaaag aactatcttc ttcaaggatg atggtaacta taagactaga
     1141 gctgaagtta agttcgaagg agatacttta gttaatagaa ttgaattgaa gggtattgat
     1201 tttaaggaag atggtaatat tttgggtcat aagttggaat ataattactt ctctgataac
     1261 gtttacatta ctgctgataa gcaaaagaac ggtattaagg cgaacttcaa gattagacat
     1321 aacattgaag atggtggtgt tcaattagct gatcattacc aacaaaacac tccaattggt
     1381 gacggtccag ttttgttgcc agataaccat tacttgtcta ctcaatctaa gttgtctaag
     1441 gatccaaacg aaaagagaga tcatatggtt ttgttggaat tcgttactgc tgcttaagcg
     1501 gccgcaccca gctttcttgt acaaagtggt gtagttaatt catgatggcc gctgcaggtc
     1561 gacctcgagg gggggcccgg tacccaattc gccctatagt gagtcgtatt acgcgcggat
     1621 ccagctttgg acttcttcgc cagaggtttg gtcaagtctc caatcaaggt tgtcggcttg
     1681 tctaccttgc cagaaattta cgaaaagatg gaaaagggtc aaatcgttgg tagatacgtt
     1741 gttgacactt ctaaataagc gaatttctta tgatttatga tttttattat taaataagtt
     1801 ataaaaaaaa taagtgtata caaattttaa agtgactctt aggttttaaa acgaaaattc
     1861 ttattcttga gtaactcttt cctgtaggtc aggttgcttt ctcaggtata gcatgaggtc
     1921 gctcttattg accacacctc taccggcatg ccaattcact ggccgtcgtt ttacaacgtc
     1981 gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg
     2041 ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
     2101 tgaatggcga atggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     2161 accgcataat cggatcgtac ttgttaccca tcattgaatt ttgaacatcc gaacctggga
     2221 gttttccctg aaacagatag tatatttgaa cctgtataat aatatatagt ctagcgcttt
     2281 acggaagaca atgtatgtat ttcggttcct ggagaaacta ttgcatctat tgcataggta
     2341 atcttgcacg tcgcatcccc ggttcatttt ctgcgtttcc atcttgcact tcaatagcat
     2401 atctttgtta acgaagcatc tgtgcttcat tttgtagaac aaaaatgcaa cgcgagagcg
     2461 ctaatttttc aaacaaagaa tctgagctgc atttttacag aacagaaatg caacgcgaaa
     2521 gcgctatttt accaacgaag aatctgtgct tcatttttgt aaaacaaaaa tgcaacgcga
     2581 gagcgctaat ttttcaaaca aagaatctga gctgcatttt tacagaacag aaatgcaacg
     2641 cgagagcgct attttaccaa caaagaatct atacttcttt tttgttctac aaaaatgcat
     2701 cccgagagcg ctatttttct aacaaagcat cttagattac tttttttctc ctttgtgcgc
     2761 tctataatgc agtctcttga taactttttg cactgtaggt ccgttaaggt tagaagaagg
     2821 ctactttggt gtctattttc tcttccataa aaaaagcctg actccacttc ccgcgtttac
     2881 tgattactag cgaagctgcg ggtgcatttt ttcaagataa aggcatcccc gattatattc
     2941 tataccgatg tggattgcgc atactttgtg aacagaaagt gatagcgttg atgattcttc
     3001 attggtcaga aaattatgaa cggtttcttc tattttgtct ctatatacta cgtataggaa
     3061 atgtttacat tttcgtattg ttttcgattc actctatgaa tagttcttac tacaattttt
     3121 ttgtctaaag agtaatacta gagataaaca taaaaaatgt agaggtcgag tttagatgca
     3181 agttcaagga gcgaaaggtg gatgggtagg ttatataggg atatagcaca gagatatata
     3241 gcaaagagat acttttgagc aatgtttgtg gaagcggtat tcgcaatatt ttagtagctc
     3301 gttacagtcc ggtgcgtttt tggttttttg aaagtgcgtc ttcagagcgc ttttggtttt
     3361 caaaagcgct ctgaagttcc tatactttct agctagagaa taggaacttc ggaataggaa
     3421 cttcaaagcg tttccgaaaa cgagcgcttc cgaaaatgca acgcgagctg cgcacataca
     3481 gctcactgtt cacgtcgcac ctatatctgc gtgttgcctg tatatatata tacatgagaa
     3541 gaacggcata gtgcgtgttt atgcttaaat gcgtacttat atgcgtctat ttatgtagga
     3601 tgaaaggtag tctagtacct cctgtgatat tatcccattc catgcggggt atcgtatgct
     3661 tccttcagca ctacccttta gctgttctat atgctgccac tcctcaattg gattagtctc
     3721 atccttcaat gctatcattt cctttgatat tggatcgatc cgatgataag ctgtcaaaca
     3781 tgagaattgg gtaataactg atataattaa attgaagctc taatttgtga gtttagtata
     3841 catgcattta cttataatac agttttttag ttttgctggc cgcatcttct caaatatgct
     3901 tcccagcctg cttttctgta acgttcaccc tctaccttag catcccttcc ctttgcaaat
     3961 agtcctcttc caacaataat aatgtcagat cctgtagaga ccacatcatc cacggttcta
     4021 tactgttgac ccaatgcgtc tcccttgtca tctaaaccca caccgggtgt cataatcaac
     4081 caatcgtaac cttcatctct tccacccatg tctctttgag caataaagcc gataacaaaa
     4141 tctttgtcgc tcttcgcaat gtcaacagta cccttagtat attctccagt agatagggag
     4201 cccttgcatg acaattctgc taacatcaaa aggcctctag gttcctttgt tacttcttct
     4261 gccgcctgct tcaaaccgct aacaatacct gggcccacca caccgtgtgc attcgtaatg
     4321 tctgcccatt ctgctattct gtatacaccc gcagagtact gcaatttgac tgtattacca
     4381 atgtcagcaa attttctgtc ttcgaagagt aaaaaattgt acttggcgga taatgccttt
     4441 agcggcttaa ctgtgccctc catggaaaaa tcagtcaaga tatccacatg tgtttttagt
     4501 aaacaaattt tgggacctaa tgcttcaact aactccagta attccttggt ggtacgaaca
     4561 tccaatgaag cacacaagtt tgtttgcttt tcgtgcatga tattaaatag cttggcagca
     4621 acaggactag gatgagtagc agcacgttcc ttatatgtag ctttcgacat gatttatctt
     4681 cgtttcctgc atgtttttgt tctgtgcagt tgggttaaga atactgggca atttcatgtt
     4741 tcttcaacac tacatatgcg tatatatacc aatctaagtc tgtgctcctt ccttcgttct
     4801 tccttctgtt cggagattac cgaatcaaaa aaatttcaag gaaaccgaaa tcaaaaaaaa
     4861 gaataaaaaa aaaatgatga attgaaaagc taattcttga agacgaaagg gcctcgtgat
     4921 acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac
     4981 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     5041 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     5101 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     5161 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     5221 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     5281 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     5341 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     5401 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     5461 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     5521 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     5581 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     5641 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     5701 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     5761 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     5821 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     5881 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     5941 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     6001 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     6061 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     6121 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     6181 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     6241 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     6301 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     6361 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     6421 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     6481 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     6541 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     6601 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     6661 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     6721 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     6781 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     6841 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     6901 agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg
     6961 gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta
     7021 gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg
     7081 aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagct
     7141 taccgcatca ggaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
     7201 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     7261 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     7321 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
     7381 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
     7441 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
     7501 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
     7561 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc attcgc
//
