LOCUS       sequence-334670-        8414 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of SweetTrac1 biosensor in yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8414)
  AUTHORS   Park J, Chavez TM, Guistwhite JA, Gwon S, Frommer WB, Cheung LS
  TITLE     Development and quantitative analysis of a biosensor based on the 
            Arabidopsis SWEET1 sugar transporter.
  JOURNAL   Proc Natl Acad Sci U S A. 2022 Jan 25;119(4). pii: 2119183119. doi: 
            10.1073/pnas.2119183119.
  PUBMED    35046045
REFERENCE   2  (bases 1 to 8414)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2022-01-25"; volume: "119(4). pii: 
            2119183119. doi"; pages: " 10.1073/pnas.2119183119"
COMMENT     Sequence Label: pDRf1-SweetTrac1
FEATURES             Location/Qualifiers
     source          1..8414
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    798..822
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2302..2326)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2388..2407)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2389..2407)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      2555..2742
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2610..2634)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(2754..2771)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2754..2770)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2763..2785)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3206..4552
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             complement(4663..5466)
                     /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(5467..5682)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(5707..5725)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5793..5897
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5898..5966,5967..6758)
                     /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(6116..6135)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6929..7517
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7418..7437
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7671..7688
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7805..7826
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7841..7846,7847..7864,7865..7871)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7879..7895
                     /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     7884..7906
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7903..7919
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7903..7919
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     rep_origin      complement(7949..8404)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8086..8107)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8298..8317
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 agcttcctga aacggagaaa cataaacagg cattgctggg atcacccata catcactctg
       61 ttttgcctga ccttttccgg taatttgaaa acaaacccgg tctcgaagcg gagatccggc
      121 gataattacc gcagaaataa acccatacac gagacgtaga accagccgca catggccgga
      181 gaaactcctg cgagaatttc gtaaactcgc gcgcattgca tctgtatttc ctaatgcggc
      241 acttccaggc ctcgatcgag accgtttatc cattgctttt ttgttgtctt tttccctcgt
      301 tcacagaaag tctgaagaag ctatagtaga actatgagct ttttttgttt ctgttttcct
      361 tttttttttt tttacctctg tggaaattgt tactctcaca ctctttagtt cgtttgtttg
      421 ttttgtttat tccaattatg accggtgacg aaacgtggtc gatggtgggt accgcttatg
      481 ctcccctcca ttagtttcga ttatataaaa aggccaaata ttgtattatt ttcaaatgtc
      541 ctatcattat cgtctaacat ctaatttctc ttaaattttt tctctttctt tcctataaca
      601 ccaatagtga aaatcttttt ttcttctata tctacaaaaa cttttttttt ctatcaacct
      661 cgttgataaa ttttttcttt aacaatcgtt aataattaat taattggaaa ataaccattt
      721 tttctctctt ttatacacac attcaaaaga aagaaaaaaa atatacccca gcctcgacta
      781 gtgaattcgc ggccatcaca agtttgtaca aaaaagcagg cttcatgaac atcgctcaca
      841 ctatcttcgg cgttttcgga aatgcaactg ctctgtttct tttcttggct ccttcgataa
      901 cattcaagag aatcatcaag aacaaatcca ctgaacaatt ctctggtatc ccttatccaa
      961 tgactctcct caactgtctc ctctctgctt ggtatggact tccctttgtg tcaaaagaca
     1021 acacacttgt gagcacaatc aatggcacag gagcagtgat cgaaacagtc tacgtgttaa
     1081 tctttctttt ctacgcacca aaggatggac agaacgtgta tattaccgcg gataaacaga
     1141 aaaacggcat taaagcgaac tttaccgtgc gccataacgt ggaagatggc agcgtgcagc
     1201 tggcggatca ttatcagcag aacaccccga ttggcgatgg cccggtgctg ctgccggata
     1261 accattatct gagcacccag accaagctga gcaaagatcc gaacgaaaaa cgcgatcaca
     1321 tggtgctgct ggaatttgtg accgcagcgg gcattacaca cggcatggat gaactgtatg
     1381 gcggcaccgg cggcagcgcg agccagggcg aagaactgtt taccggcgtg gtgccgattc
     1441 tggtggaact ggatggcgat gtgaacggcc ataaatttag cgtgcgcggc gaaggcgaag
     1501 gcgatgcgac cattggcaaa ctgaccctga aatttatttc caccaccggc aaactaccgg
     1561 tgccgtggcc gaccctggtg accaccttaa cctatggcgt gcagtgcttt agccgctatc
     1621 cggatcatat gaaacgccat gattttttta aaagcgcgat gccggaaggc tatgtgcagg
     1681 aacgcaccat tagctttaaa gatgatggca aatataaaac ccgcgcggtg gtgaaatttg
     1741 aaggcgatac cctggtgaac cgcattgaac tgaaaggcac cgattttaaa gaagatggca
     1801 acattctggg gcataaactg gaatataacc ttactagaaa ggagaaaatt aagatctttg
     1861 gtatcttctc ttgcgttttg gctgtattcg caacagtagc tctcgtctct ctctttgctc
     1921 ttcaaggaaa cggtagaaaa ctcttttgtg gtctcgcagc tactgttttc tccatcatca
     1981 tgtacgcttc tccactctcc atcatgagat tggtggtaaa gacgaagagt gtagagttta
     2041 tgccattctt tttgtcactc tttgtcttcc tctgtggtac ttcgtggttc gtctatggtc
     2101 taatcggtcg tgaccctttt gttgcaatcc caaatgggtt tggatgtgca ttagggacac
     2161 tgcaattgat actatacttc atctactgtg gaaacaaagg agagaaatct gcagatgctc
     2221 agaaagatga gaagtcagtg gagatgaaag atgatgagaa gaagcagaat gtggttaatg
     2281 gaaagcaaga ccttcaagtt gacccagctt tcttgtacaa agtggtgtag ttaattcatg
     2341 atggccgctg caggtcgacc tcgagggggg gcccggtacc caattcgccc tatagtgagt
     2401 cgtattacgc gcggatccag ctttggactt cttcgccaga ggtttggtca agtctccaat
     2461 caaggttgtc ggcttgtcta ccttgccaga aatttacgaa aagatggaaa agggtcaaat
     2521 cgttggtaga tacgttgttg acacttctaa ataagcgaat ttcttatgat ttatgatttt
     2581 tattattaaa taagttataa aaaaaataag tgtatacaaa ttttaaagtg actcttaggt
     2641 tttaaaacga aaattcttat tcttgagtaa ctctttcctg taggtcaggt tgctttctca
     2701 ggtatagcat gaggtcgctc ttattgacca cacctctacc ggcatgccaa ttcactggcc
     2761 gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca
     2821 gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
     2881 caacagttgc gcagcctgaa tggcgaatgg cgcctgatgc ggtattttct ccttacgcat
     2941 ctgtgcggta tttcacaccg cataatcgga tcgtacttgt tacccatcat tgaattttga
     3001 acatccgaac ctgggagttt tccctgaaac agatagtata tttgaacctg tataataata
     3061 tatagtctag cgctttacgg aagacaatgt atgtatttcg gttcctggag aaactattgc
     3121 atctattgca taggtaatct tgcacgtcgc atccccggtt cattttctgc gtttccatct
     3181 tgcacttcaa tagcatatct ttgttaacga agcatctgtg cttcattttg tagaacaaaa
     3241 atgcaacgcg agagcgctaa tttttcaaac aaagaatctg agctgcattt ttacagaaca
     3301 gaaatgcaac gcgaaagcgc tattttacca acgaagaatc tgtgcttcat ttttgtaaaa
     3361 caaaaatgca acgcgagagc gctaattttt caaacaaaga atctgagctg catttttaca
     3421 gaacagaaat gcaacgcgag agcgctattt taccaacaaa gaatctatac ttcttttttg
     3481 ttctacaaaa atgcatcccg agagcgctat ttttctaaca aagcatctta gattactttt
     3541 tttctccttt gtgcgctcta taatgcagtc tcttgataac tttttgcact gtaggtccgt
     3601 taaggttaga agaaggctac tttggtgtct attttctctt ccataaaaaa agcctgactc
     3661 cacttcccgc gtttactgat tactagcgaa gctgcgggtg cattttttca agataaaggc
     3721 atccccgatt atattctata ccgatgtgga ttgcgcatac tttgtgaaca gaaagtgata
     3781 gcgttgatga ttcttcattg gtcagaaaat tatgaacggt ttcttctatt ttgtctctat
     3841 atactacgta taggaaatgt ttacattttc gtattgtttt cgattcactc tatgaatagt
     3901 tcttactaca atttttttgt ctaaagagta atactagaga taaacataaa aaatgtagag
     3961 gtcgagttta gatgcaagtt caaggagcga aaggtggatg ggtaggttat atagggatat
     4021 agcacagaga tatatagcaa agagatactt ttgagcaatg tttgtggaag cggtattcgc
     4081 aatattttag tagctcgtta cagtccggtg cgtttttggt tttttgaaag tgcgtcttca
     4141 gagcgctttt ggttttcaaa agcgctctga agttcctata ctttctagct agagaatagg
     4201 aacttcggaa taggaacttc aaagcgtttc cgaaaacgag cgcttccgaa aatgcaacgc
     4261 gagctgcgca catacagctc actgttcacg tcgcacctat atctgcgtgt tgcctgtata
     4321 tatatataca tgagaagaac ggcatagtgc gtgtttatgc ttaaatgcgt acttatatgc
     4381 gtctatttat gtaggatgaa aggtagtcta gtacctcctg tgatattatc ccattccatg
     4441 cggggtatcg tatgcttcct tcagcactac cctttagctg ttctatatgc tgccactcct
     4501 caattggatt agtctcatcc ttcaatgcta tcatttcctt tgatattgga tcgatccgat
     4561 gataagctgt caaacatgag aattgggtaa taactgatat aattaaattg aagctctaat
     4621 ttgtgagttt agtatacatg catttactta taatacagtt ttttagtttt gctggccgca
     4681 tcttctcaaa tatgcttccc agcctgcttt tctgtaacgt tcaccctcta ccttagcatc
     4741 ccttcccttt gcaaatagtc ctcttccaac aataataatg tcagatcctg tagagaccac
     4801 atcatccacg gttctatact gttgacccaa tgcgtctccc ttgtcatcta aacccacacc
     4861 gggtgtcata atcaaccaat cgtaaccttc atctcttcca cccatgtctc tttgagcaat
     4921 aaagccgata acaaaatctt tgtcgctctt cgcaatgtca acagtaccct tagtatattc
     4981 tccagtagat agggagccct tgcatgacaa ttctgctaac atcaaaaggc ctctaggttc
     5041 ctttgttact tcttctgccg cctgcttcaa accgctaaca atacctgggc ccaccacacc
     5101 gtgtgcattc gtaatgtctg cccattctgc tattctgtat acacccgcag agtactgcaa
     5161 tttgactgta ttaccaatgt cagcaaattt tctgtcttcg aagagtaaaa aattgtactt
     5221 ggcggataat gcctttagcg gcttaactgt gccctccatg gaaaaatcag tcaagatatc
     5281 cacatgtgtt tttagtaaac aaattttggg acctaatgct tcaactaact ccagtaattc
     5341 cttggtggta cgaacatcca atgaagcaca caagtttgtt tgcttttcgt gcatgatatt
     5401 aaatagcttg gcagcaacag gactaggatg agtagcagca cgttccttat atgtagcttt
     5461 cgacatgatt tatcttcgtt tcctgcatgt ttttgttctg tgcagttggg ttaagaatac
     5521 tgggcaattt catgtttctt caacactaca tatgcgtata tataccaatc taagtctgtg
     5581 ctccttcctt cgttcttcct tctgttcgga gattaccgaa tcaaaaaaat ttcaaggaaa
     5641 ccgaaatcaa aaaaaagaat aaaaaaaaaa tgatgaattg aaaagctaat tcttgaagac
     5701 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     5761 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     5821 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     5881 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     5941 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     6001 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     6061 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     6121 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     6181 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     6241 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     6301 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     6361 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     6421 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     6481 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     6541 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     6601 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     6661 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     6721 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     6781 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     6841 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     6901 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     6961 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     7021 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     7081 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     7141 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     7201 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     7261 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     7321 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     7381 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     7441 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     7501 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     7561 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     7621 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     7681 tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
     7741 ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg
     7801 caattaatgt gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg
     7861 ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc
     7921 atgattacgc caagcttacc gcatcaggaa attgtaagcg ttaatatttt gttaaaattc
     7981 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
     8041 ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag
     8101 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
     8161 gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa
     8221 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
     8281 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
     8341 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
     8401 gcgtccattc gcca
//
