LOCUS       sequence-437723-        8431 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of ymTq2-T2A-ymScarletl in yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8431)
  TITLE     Teusink lab plasmids
REFERENCE   2  (bases 1 to 8431)
  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"
COMMENT     Sequence Label: pDRF1-GW ymTq2-T2A-ymScarletl
FEATURES             Location/Qualifiers
     source          1..8431
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    800..824
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1548..1601
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     protein_bind    complement(2321..2345)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2407..2426)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2408..2426)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      2574..2761
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2629..2653)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(2773..2790)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2773..2789)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2782..2804)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3225..4571
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             complement(4682..5485)
                     /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(5486..5701)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(5726..5744)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5812..5916
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5917..5985,5986..6777)
                     /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(6135..6154)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6948..7536
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7437..7456
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7690..7707
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7824..7845
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7860..7865,7866..7883,7884..7890)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7898..7914
                     /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     7903..7925
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7922..7938
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7922..7938
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     rep_origin      complement(7968..8423)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8105..8126)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8317..8336
                     /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 tagtgaattc gcggccatca caagtttgta caaaaaagca ggctgctagc atggttagta
      841 aaggtgaaga attgttcact ggtgtagtgc caattttagt agaattagat ggtgacgtta
      901 acggtcataa gttctctgta agcggtgaag gtgaaggtga cgcaacatac ggtaaattaa
      961 ctttgaaatt catttgtact acaggtaaat taccagttcc atggccaact ttggttacta
     1021 ctttgtcttg gggtgttcaa tgtttcgcta gatacccaga tcatatgaag caacatgatt
     1081 tctttaagtc tgctatgcca gaaggttacg ttcaagaaag aactatcttc ttcaaggatg
     1141 atggtaacta taagactaga gctgaagtta agttcgaagg agatacttta gttaatagaa
     1201 ttgaattgaa gggtattgat tttaaggaag atggtaatat tttgggtcat aagttggaat
     1261 ataattactt ctctgataac gtttacatta ctgctgataa gcaaaagaac ggtattaagg
     1321 cgaacttcaa gattagacat aacattgaag atggtggtgt tcaattagct gatcattacc
     1381 aacaaaacac tccaattggt gacggtccag ttttgttgcc agataaccat tacttgtcta
     1441 ctcaatctaa gttgtctaag gatccaaacg aaaagagaga tcatatggtt ttgttggaat
     1501 tcgttactgc tgctgggatc actttaggta tggatgaatt gtataaagag ggcagaggaa
     1561 gtcttctaac atgcggtgac gtggaggaga atcccggccc tggggatccc gggatggtta
     1621 gtaaaggtga agctgttata aaagaattca tgaggtttaa agttcatatg gaaggttcaa
     1681 tgaatggtca tgaatttgaa attgaaggtg aaggtgaagg tagaccatat gaaggtacac
     1741 aaactgctaa attgaaagtt actaaaggtg gtccattacc attttcttgg gatattttgt
     1801 ctccacaatt catgtatggt tctagagctt ttattaagca tccagctgat attccagatt
     1861 attataaaca atcttttcct gaaggtttta aatgggaaag agttatgaat ttcgaagatg
     1921 gtggtgctgt tactgttact caagatactt ctttggaaga tggtacttta atctataaag
     1981 ttaaattgag aggtactaat tttccaccag atggtccagt tatgcaaaag aaaactatgg
     2041 gttgggaagc atctactgaa agattgtatc cagaagatgg tgttttgaaa ggagacatta
     2101 agatggcttt gagattgaaa gatggtggtc ggtacctggc cgacttcaag accacctata
     2161 aagctaaaaa accagttcaa atgccaggtg catataatgt tgatagaaag ttagatataa
     2221 cgtcgcataa cgaggactat actgtagttg aacaatatga acgtagtgaa ggtagacata
     2281 gtaccggagg aatggatgaa ttgtataaat aagcggccgc acccagcttt cttgtacaaa
     2341 gtggtgtagt taattcatga tggccgctgc aggtcgacct cgaggggggg cccggtaccc
     2401 aattcgccct atagtgagtc gtattacgcg cggatccagc tttggacttc ttcgccagag
     2461 gtttggtcaa gtctccaatc aaggttgtcg gcttgtctac cttgccagaa atttacgaaa
     2521 agatggaaaa gggtcaaatc gttggtagat acgttgttga cacttctaaa taagcgaatt
     2581 tcttatgatt tatgattttt attattaaat aagttataaa aaaaataagt gtatacaaat
     2641 tttaaagtga ctcttaggtt ttaaaacgaa aattcttatt cttgagtaac tctttcctgt
     2701 aggtcaggtt gctttctcag gtatagcatg aggtcgctct tattgaccac acctctaccg
     2761 gcatgccaat tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac
     2821 ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc
     2881 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gcctgatgcg
     2941 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc ataatcggat cgtacttgtt
     3001 acccatcatt gaattttgaa catccgaacc tgggagtttt ccctgaaaca gatagtatat
     3061 ttgaacctgt ataataatat atagtctagc gctttacgga agacaatgta tgtatttcgg
     3121 ttcctggaga aactattgca tctattgcat aggtaatctt gcacgtcgca tccccggttc
     3181 attttctgcg tttccatctt gcacttcaat agcatatctt tgttaacgaa gcatctgtgc
     3241 ttcattttgt agaacaaaaa tgcaacgcga gagcgctaat ttttcaaaca aagaatctga
     3301 gctgcatttt tacagaacag aaatgcaacg cgaaagcgct attttaccaa cgaagaatct
     3361 gtgcttcatt tttgtaaaac aaaaatgcaa cgcgagagcg ctaatttttc aaacaaagaa
     3421 tctgagctgc atttttacag aacagaaatg caacgcgaga gcgctatttt accaacaaag
     3481 aatctatact tcttttttgt tctacaaaaa tgcatcccga gagcgctatt tttctaacaa
     3541 agcatcttag attacttttt ttctcctttg tgcgctctat aatgcagtct cttgataact
     3601 ttttgcactg taggtccgtt aaggttagaa gaaggctact ttggtgtcta ttttctcttc
     3661 cataaaaaaa gcctgactcc acttcccgcg tttactgatt actagcgaag ctgcgggtgc
     3721 attttttcaa gataaaggca tccccgatta tattctatac cgatgtggat tgcgcatact
     3781 ttgtgaacag aaagtgatag cgttgatgat tcttcattgg tcagaaaatt atgaacggtt
     3841 tcttctattt tgtctctata tactacgtat aggaaatgtt tacattttcg tattgttttc
     3901 gattcactct atgaatagtt cttactacaa tttttttgtc taaagagtaa tactagagat
     3961 aaacataaaa aatgtagagg tcgagtttag atgcaagttc aaggagcgaa aggtggatgg
     4021 gtaggttata tagggatata gcacagagat atatagcaaa gagatacttt tgagcaatgt
     4081 ttgtggaagc ggtattcgca atattttagt agctcgttac agtccggtgc gtttttggtt
     4141 ttttgaaagt gcgtcttcag agcgcttttg gttttcaaaa gcgctctgaa gttcctatac
     4201 tttctagcta gagaatagga acttcggaat aggaacttca aagcgtttcc gaaaacgagc
     4261 gcttccgaaa atgcaacgcg agctgcgcac atacagctca ctgttcacgt cgcacctata
     4321 tctgcgtgtt gcctgtatat atatatacat gagaagaacg gcatagtgcg tgtttatgct
     4381 taaatgcgta cttatatgcg tctatttatg taggatgaaa ggtagtctag tacctcctgt
     4441 gatattatcc cattccatgc ggggtatcgt atgcttcctt cagcactacc ctttagctgt
     4501 tctatatgct gccactcctc aattggatta gtctcatcct tcaatgctat catttccttt
     4561 gatattggat cgatccgatg ataagctgtc aaacatgaga attgggtaat aactgatata
     4621 attaaattga agctctaatt tgtgagttta gtatacatgc atttacttat aatacagttt
     4681 tttagttttg ctggccgcat cttctcaaat atgcttccca gcctgctttt ctgtaacgtt
     4741 caccctctac cttagcatcc cttccctttg caaatagtcc tcttccaaca ataataatgt
     4801 cagatcctgt agagaccaca tcatccacgg ttctatactg ttgacccaat gcgtctccct
     4861 tgtcatctaa acccacaccg ggtgtcataa tcaaccaatc gtaaccttca tctcttccac
     4921 ccatgtctct ttgagcaata aagccgataa caaaatcttt gtcgctcttc gcaatgtcaa
     4981 cagtaccctt agtatattct ccagtagata gggagccctt gcatgacaat tctgctaaca
     5041 tcaaaaggcc tctaggttcc tttgttactt cttctgccgc ctgcttcaaa ccgctaacaa
     5101 tacctgggcc caccacaccg tgtgcattcg taatgtctgc ccattctgct attctgtata
     5161 cacccgcaga gtactgcaat ttgactgtat taccaatgtc agcaaatttt ctgtcttcga
     5221 agagtaaaaa attgtacttg gcggataatg cctttagcgg cttaactgtg ccctccatgg
     5281 aaaaatcagt caagatatcc acatgtgttt ttagtaaaca aattttggga cctaatgctt
     5341 caactaactc cagtaattcc ttggtggtac gaacatccaa tgaagcacac aagtttgttt
     5401 gcttttcgtg catgatatta aatagcttgg cagcaacagg actaggatga gtagcagcac
     5461 gttccttata tgtagctttc gacatgattt atcttcgttt cctgcatgtt tttgttctgt
     5521 gcagttgggt taagaatact gggcaatttc atgtttcttc aacactacat atgcgtatat
     5581 ataccaatct aagtctgtgc tccttccttc gttcttcctt ctgttcggag attaccgaat
     5641 caaaaaaatt tcaaggaaac cgaaatcaaa aaaaagaata aaaaaaaaat gatgaattga
     5701 aaagctaatt cttgaagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
     5761 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     5821 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     5881 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     5941 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     6001 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     6061 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     6121 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     6181 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     6241 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     6301 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     6361 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     6421 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     6481 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     6541 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     6601 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     6661 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     6721 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     6781 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     6841 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     6901 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     6961 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     7021 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     7081 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     7141 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     7201 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     7261 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     7321 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     7381 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     7441 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     7501 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     7561 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     7621 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     7681 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct
     7741 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
     7801 gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct
     7861 ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac
     7921 acaggaaaca gctatgacca tgattacgcc aagcttaccg catcaggaaa ttgtaagcgt
     7981 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
     8041 ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt
     8101 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
     8161 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt
     8221 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
     8281 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
     8341 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
     8401 taatgcgccg ctacagggcg cgtccattcg c
//
