LOCUS       sequence-326051-        8433 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  FRET fusion between ymTq2 and ymNeongreen for use in yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8433)
  TITLE     Teusink unpublished plasmids
REFERENCE   2  (bases 1 to 8433)
  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: ymTq2-ymNeongreen pDRF1-GW
FEATURES             Location/Qualifiers
     source          1..8433
                     /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"
     regulatory      1598..1607
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      1859..1868
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(2323..2347)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2409..2428)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2410..2428)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      2576..2763
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2631..2655)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(2775..2792)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2775..2791)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2784..2806)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3227..4573
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             complement(4684..5487)
                     /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(5488..5703)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(5728..5746)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5814..5918
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5919..5987,5988..6779)
                     /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(6137..6156)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6950..7538
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7439..7458
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7692..7709
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7826..7847
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7862..7867,7868..7885,7886..7892)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7900..7916
                     /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     7905..7927
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7924..7940
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7924..7940
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     rep_origin      complement(7970..8425)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8107..8128)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8319..8338
                     /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 actagtaagc
      841 ttttaattaa atggttagta aaggtgaaga attgttcact ggtgtagtgc caattttagt
      901 agaattagat ggtgacgtta acggtcataa gttctctgta agcggtgaag gtgaaggtga
      961 cgcaacatac ggtaaattaa ctttgaaatt catttgtact acaggtaaat taccagttcc
     1021 atggccaact ttggttacta ctttgtcttg gggtgttcaa tgtttcgcta gatacccaga
     1081 tcatatgaag caacatgatt tctttaagtc tgctatgcca gaaggttacg ttcaagaaag
     1141 aactatcttc ttcaaggatg atggtaacta taagactaga gctgaagtta agttcgaagg
     1201 agatacttta gttaatagaa ttgaattgaa gggtattgat tttaaggaag atggtaatat
     1261 tttgggtcat aagttggaat ataattactt ctctgataac gtttacatta ctgctgataa
     1321 gcaaaagaac ggtattaagg cgaacttcaa gattagacat aacattgaag atggtggtgt
     1381 tcaattagct gatcattacc aacaaaacac tccaattggt gacggtccag ttttgttgcc
     1441 agataaccat tacttgtcta ctcaatctaa gttgtctaag gatccaaacg aaaagagaga
     1501 tcatatggtt ttgttggaat tcgttactgc tgctgggatc actttaggta tggatgaatt
     1561 gtataaatcc ggactcagat cttcggatcc accggtcgcc accatggtct ctaagggtga
     1621 agaagataac atggcttctt tgccagccac ccacgagttg cacatcttcg gttccattaa
     1681 cggtgtcgac ttcgatatgg tcggtcaagg taccggtaac ccaaacgacg gttacgaaga
     1741 attgaacttg aagtctacca agggcgactt gcagttctcc ccttggattt tggttccaca
     1801 catcggttac ggtttccacc aatacttgcc atacccagac ggtatgtccc cattccaagc
     1861 cgccatggtt gacggttctg gttaccaagt ccacagaacc atgcaattcg aagacggtgc
     1921 ttccttgacc gtgaactaca gatacaccta cgaaggttct cacatcaagg gcgaagccca
     1981 agttaagggt accggtttcc ctgctgacgg tccagtcatg actaactcct tgaccgctgc
     2041 tgactggtgt cgttccaaga agacctaccc aaacgataag accatcattt ccactttcaa
     2101 gtggtcctac accaccggta acggcaagag ataccgctct accgctcgta ccacctacac
     2161 cttcgccaag ccaatggctg ccaactactt gaagaaccag cctatgtacg tcttccgtaa
     2221 gaccgagctg aagcattcca agactgaatt gaacttcaag gaatggcaaa aggctttcac
     2281 cgacgtcatg ggtatggacg aattgtacaa gtaagcggcc gcacccagct ttcttgtaca
     2341 aagtggtgta gttaattcat gatggccgct gcaggtcgac ctcgaggggg ggcccggtac
     2401 ccaattcgcc ctatagtgag tcgtattacg cgcggatcca gctttggact tcttcgccag
     2461 aggtttggtc aagtctccaa tcaaggttgt cggcttgtct accttgccag aaatttacga
     2521 aaagatggaa aagggtcaaa tcgttggtag atacgttgtt gacacttcta aataagcgaa
     2581 tttcttatga tttatgattt ttattattaa ataagttata aaaaaaataa gtgtatacaa
     2641 attttaaagt gactcttagg ttttaaaacg anaattctta ttcttgagta actctttcct
     2701 gtaggtcagg ttgctttctc aggtatagca tgaggtcgct cttattgacc acacctctac
     2761 cggcatgcca attcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt
     2821 acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag
     2881 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg gcgcctgatg
     2941 cggtattttc tccttacgca tctgtgcggt atttcacacc gcataatcgg atcgtacttg
     3001 ttacccatca ttgaattttg aacatccgaa cctgggagtt ttccctgaaa cagatagtat
     3061 atttgaacct gtataataat atatagtcta gcgctttacg gaagacaatg tatgtatttc
     3121 ggttcctgga gaaactattg catctattgc ataggtaatc ttgcacgtcg catccccggt
     3181 tcattttctg cgtttccatc ttgcacttca atagcatatc tttgttaacg aagcatctgt
     3241 gcttcatttt gtagaacaaa aatgcaacgc gagagcgcta atttttcaaa caaagaatct
     3301 gagctgcatt tttacagaac agaaatgcaa cgcgaaagcg ctattttacc aacgaagaat
     3361 ctgtgcttca tttttgtaaa acaaaaatgc aacgcgagag cgctaatttt tcaaacaaag
     3421 aatctgagct gcatttttac agaacagaaa tgcaacgcga gagcgctatt ttaccaacaa
     3481 agaatctata cttctttttt gttctacaaa aatgcatccc gagagcgcta tttttctaac
     3541 aaagcatctt agattacttt ttttctcctt tgtgcgctct ataatgcagt ctcttgataa
     3601 ctttttgcac tgtaggtccg ttaaggttag aagaaggcta ctttggtgtc tattttctct
     3661 tccataaaaa aagcctgact ccacttcccg cgtttactga ttactagcga agctgcgggt
     3721 gcattttttc aagataaagg catccccgat tatattctat accgatgtgg attgcgcata
     3781 ctttgtgaac agaaagtgat agcgttgatg attcttcatt ggtcagaaaa ttatgaacgg
     3841 tttcttctat tttgtctcta tatactacgt ataggaaatg tttacatttt cgtattgttt
     3901 tcgattcact ctatgaatag ttcttactac aatttttttg tctaaagagt aatactagag
     3961 ataaacataa aaaatgtaga ggtcgagttt agatgcaagt tcaaggagcg aaaggtggat
     4021 gggtaggtta tatagggata tagcacagag atatatagca aagagatact tttgagcaat
     4081 gtttgtggaa gcggtattcg caatatttta gtagctcgtt acagtccggt gcgtttttgg
     4141 ttttttgaaa gtgcgtcttc agagcgcttt tggttttcaa aagcgctctg aagttcctat
     4201 actttctagc tagagaatag gaacttcgga ataggaactt caaagcgttt ccgaaaacga
     4261 gcgcttccga aaatgcaacg cgagctgcgc acatacagct cactgttcac gtcgcaccta
     4321 tatctgcgtg ttgcctgtat atatatatac atgagaagaa cggcatagtg cgtgtttatg
     4381 cttaaatgcg tacttatatg cgtctattta tgtaggatga aaggtagtct agtacctcct
     4441 gtgatattat cccattccat gcggggtatc gtatgcttcc ttcagcacta ccctttagct
     4501 gttctatatg ctgccactcc tcaattggat tagtctcatc cttcaatgct atcatttcct
     4561 ttgatattgg atcgatccga tgataagctg tcaaacatga gaattgggta ataactgata
     4621 taattaaatt gaagctctaa tttgtgagtt tagtatacat gcatttactt ataatacagt
     4681 tttttagttt tgctggccgc atcttctcaa atatgcttcc cagcctgctt ttctgtaacg
     4741 ttcaccctct accttagcat cccttccctt tgcaaatagt cctcttccaa caataataat
     4801 gtcagatcct gtagagacca catcatccac ggttctatac tgttgaccca atgcgtctcc
     4861 cttgtcatct aaacccacac cgggtgtcat aatcaaccaa tcgtaacctt catctcttcc
     4921 acccatgtct ctttgagcaa taaagccgat aacaaaatct ttgtcgctct tcgcaatgtc
     4981 aacagtaccc ttagtatatt ctccagtaga tagggagccc ttgcatgaca attctgctaa
     5041 catcaaaagg cctctaggtt cctttgttac ttcttctgcc gcctgcttca aaccgctaac
     5101 aatacctggg cccaccacac cgtgtgcatt cgtaatgtct gcccattctg ctattctgta
     5161 tacacccgca gagtactgca atttgactgt attaccaatg tcagcaaatt ttctgtcttc
     5221 gaagagtaaa aaattgtact tggcggataa tgcctttagc ggcttaactg tgccctccat
     5281 ggaaaaatca gtcaagatat ccacatgtgt ttttagtaaa caaattttgg gacctaatgc
     5341 ttcaactaac tccagtaatt ccttggtggt acgaacatcc aatgaagcac acaagtttgt
     5401 ttgcttttcg tgcatgatat taaatagctt ggcagcaaca ggactaggat gagtagcagc
     5461 acgttcctta tatgtagctt tcgacatgat ttatcttcgt ttcctgcatg tttttgttct
     5521 gtgcagttgg gttaagaata ctgggcaatt tcatgtttct tcaacactac atatgcgtat
     5581 atataccaat ctaagtctgt gctccttcct tcgttcttcc ttctgttcgg agattaccga
     5641 atcaaaaaaa tttcaaggaa accgaaatca aaaaaaagaa taaaaaaaaa atgatgaatt
     5701 gaaaagctaa ttcttgaaga cgaaagggcc tcgtgatacg cctattttta taggttaatg
     5761 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     5821 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     5881 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     5941 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     6001 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     6061 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     6121 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     6181 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     6241 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     6301 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     6361 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     6421 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     6481 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     6541 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     6601 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     6661 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     6721 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     6781 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     6841 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     6901 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     6961 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     7021 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     7081 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     7141 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     7201 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     7261 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     7321 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     7381 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     7441 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     7501 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     7561 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     7621 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     7681 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc
     7741 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga
     7801 aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg
     7861 ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc
     7921 acacaggaaa cagctatgac catgattacg ccaagcttac cgcatcagga aattgtaagc
     7981 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     8041 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
     8101 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
     8161 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
     8221 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
     8281 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
     8341 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
     8401 cttaatgcgc cgctacaggg cgcgtccatt cgc
//
