LOCUS       sequence-404927-        8227 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To express yeast Hxt1 hexose transporter in yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8227)
  AUTHORS   Fuhrmeister R, Streubel J
  TITLE     Functional Analysis of Plant Monosaccharide Transporters Using a 
            Simple Growth Complementation Assay in Yeast.
  JOURNAL   Bio Protoc. 2023 Aug 5;13(15):e4733. doi: 10.21769/BioProtoc.4733. 
            eCollection 2023 Aug 5.
  PUBMED    37575400
REFERENCE   2  (bases 1 to 8227)
  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: "Bio 
            Protoc."; date: "2023-08-5"; volume: "13(15):e4733. doi"; pages: " 
            10.21769/BioProtoc.4733. eCollection 2023 Aug 5"
COMMENT     Sequence Label: pJS288
FEATURES             Location/Qualifiers
     source          1..8227
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             2759..3469
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2904..2922)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     terminator      3611..3798
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(3666..3690)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     rep_origin      4424..5301
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     promoter        5310..5414
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5415..5483,5484..6275)
                     /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(5633..5652)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6487..7075
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6976..6995
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7198..8001)
                     /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(8002..8222)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
ORIGIN
        1 tggtcactct gtgaagacaa tgccgaattc ggatccggag ctcagctttg ctaaagtgca
       61 aaaagtcgtt tacgtcacca tcgctgacgg ggaaaaaaca aaacaagctt cctgaaacgg
      121 agaaacataa acaggcattg ctgggatcac ccatacatca ctctgttttg cctgaccttt
      181 tccggtaatt tgaaaacaaa cccggtttcg aagcggagat ccggcgataa ttaccgcaga
      241 aataaaccca tacacgagac gtagaaccag ccgcacatgg ccggagaaac tcctgcgaga
      301 atttcgtaaa ctcgcgcgca ttgcatctgt atttcctaat gcggcacttc caggcctcga
      361 aacctctgac atgcttttga caggaataga cattttcaga atgttatcca tatgcctttc
      421 gggttttttt ccttcctttt ccatcatgaa aaatctctca agaccgttta tccattgctt
      481 ttttgttgtc tttttccctc gttcacagaa agtctgaaga agctatagta gaactatgag
      541 ctttttttgt ttctgttttc cttttttttt ttttttacct ctgtggaaat tgttactctc
      601 acactcttta gttcgtttgt ttgttttgtt tattccaatt atgaccggtg acgaaacgtg
      661 gtcgatggtg ggtaccgctt atgctcccct ccattagttt cgattatata aaaaggccaa
      721 atattgtatt attttcaaat gtcctatcat tatcgtctaa catctaattt ctcttaaatt
      781 ttttctcttt ctttcctata acaccaatag tgaaaatctt tttttcttct atatctacaa
      841 aaactttttt tttctatcaa cctcgttgat aaattttttc tttaacaatc gttaataatt
      901 aattaattgg aaaataacca ttttttctct cttttataca cacattcaaa agaaagaaaa
      961 aaaatatacc ccagctagtt aaagaaaatc attgaaaaga ataagaagat aagaaagatt
     1021 taattatcaa acaatatcaa taatgaattc aactcccgat ctaatatctc ctcagaaatc
     1081 caattcatcc aactcatatg aattggaatc tggtcgttca aaggccatga atactccaga
     1141 aggtaaaaat gaaagttttc acgacaactt aagtgaaagt caagtgcaac ccgccgttgc
     1201 ccctccaaac accggaaaag gtgtctacgt aacggtttct atctgttgtg ttatggttgc
     1261 tttcggtggt ttcatatttg gatgggatac tggtaccatt tctggttttg ttgctcaaac
     1321 tgattttcta agaagatttg gtatgaagca ccacgacggt agtcattact tgtccaaggt
     1381 gagaactggt ttaattgtct ctatttttaa cattggttgt gccattggtg gtatcgtctt
     1441 agccaagcta ggtgatatgt atggtcgtag aatcggtttg attgtcgttg tagtaatcta
     1501 cactatcggt atcattattc aaatagcctc gatcaacaag tggtaccaat atttcattgg
     1561 tagaattatc tctggtttag gtgtcggtgg tatcacagtt ttatctccca tgctaatatc
     1621 tgaggtcgcc cccagtgaaa tgagaggcac cttggtttca tgttaccaag tcatgattac
     1681 tttaggtatt ttcttaggtt actgtaccaa ttttggtacc aagaattact caaactctgt
     1741 ccaatggaga gttccattag gtttgtgttt cgcctgggcc ttatttatga ttggtggtat
     1801 gatgtttgtt cctgaatctc cacgttattt ggttgaagct ggcagaatcg acgaagccag
     1861 ggcttcttta gctaaagtta acaaatgccc acctgaccat ccatacattc aatatgagtt
     1921 ggaaactatc gaagccagtg tcgaagaaat gagagccgct ggtactgcat cttggggcga
     1981 attattcact ggtaaaccag ccatgtttca acgtactatg atgggtatca tgattcaatc
     2041 tctacaacaa ttaactggtg ataactattt cttctactac ggtaccattg ttttccaggc
     2101 tgtcggttta agtgactctt ttgaaacttc tattgtcttt ggtgtcgtca acttcttctc
     2161 cacttgttgt tctctgtaca ccgttgaccg ttttggccgt cgtaactgtt tgatgtgggg
     2221 tgctgtcggt atggtctgct gttatgttgt ctatgcctct gttggtgtta ccagattatg
     2281 gccaaacggt caagatcaac catcttcaaa gggtgctggt aactgtatga ttgttttcgc
     2341 atgtttctac attttctgtt tcgctactac ctgggcccca attgcttacg ttgttatttc
     2401 agaatgtttc ccattaagag tcaaatccaa gtgtatgtct attgccagtg ctgctaactg
     2461 gatctggggt ttcttgatta gtttcttcac cccatttatt actggtgcca tcaacttcta
     2521 ctacggttac gttttcatgg gctgtatggt tttcgcttac ttttacgtct ttttcttcgt
     2581 tccagaaact aaaggtttat cattagaaga agttaatgat atgtacgccg aaggtgttct
     2641 accatggaaa tcagcttcct gggttccagt atccaagaga ggcgctgact acaacgctga
     2701 tgacctaatg catgatgacc aaccatttta caagagtttg tttagcagga aattcggcat
     2761 ggtgagcaag ggcgaggagg ataacatggc catcatcaag gagttcatgc gcttcaaggt
     2821 gcacatggag ggctccgtga acggccacga gttcgagatc gagggcgagg gcgagggccg
     2881 cccctacgag ggcacccaga ccgccaagct gaaggtgacc aagggtggcc ccctgccctt
     2941 cgcctgggac atcctgtccc ctcagttcat gtacggctcc aaggcctacg tgaagcaccc
     3001 cgccgacatc cccgactact tgaagctgtc cttccccgag ggcttcaagt gggagcgcgt
     3061 gatgaacttc gaggacggcg gcgtggtgac cgtgacccag gactcctccc tgcaggacgg
     3121 cgagttcatc tacaaggtga agctgcgcgg caccaacttc ccctccgacg gccccgtaat
     3181 gcagaagaaa accatgggct gggaggcctc ctccgagcgg atgtaccccg aggacggcgc
     3241 cctgaagggc gagatcaagc agaggctgaa gctgaaggac ggcggccact acgacgctga
     3301 ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg ccaggcgcct acaacgtcaa
     3361 catcaagttg gacatcacct cccacaacga ggactacacc atcgtggaac agtacgaacg
     3421 cgccgagggc cgccactcca ccggcggcat ggacgagctg tacaagtaag cttaagcttt
     3481 ggacttcttc gccagaggtt tggtcaagtc tccaatcaag gttgtcggct tgtctacctt
     3541 gccagaaatt tacgaaaaga tggaaaaggg tcaaatcgtt ggtagatacg ttgttgacac
     3601 ttctaaataa gcgaatttct tatgatttat gatttttatt attaaataag ttataaaaaa
     3661 aataagtgta tacaaatttt aaagtgactc ttaggtttta aaacgaaaat tcttattctt
     3721 gagtaactct ttcctgtagg tcaggttgct ttctcaggta tagcatgagg tcgctcttat
     3781 tgaccacacc tctaccggcc cgctgcaatt gtcttctgca cgaagtggcc gttatcgatg
     3841 ataagctgtc aaacatgaga attaattcca cggactatag actataccta gtatactccg
     3901 tctactgtac gatacacttc cgctcaggtc cttgtccttt aacgaggcct taccactctt
     3961 ttgttactct attgatccag ctcagcaaag gcagtgtgat ctaagattct atcttcgcga
     4021 tgtagtaaaa ctagctagac cgagaaagag actagaaatg caaaaggcac ttctacaatg
     4081 gctgccatca ttattatgcg atgtgacgct gcagcttctc aatgatattc gaatacgctt
     4141 tgaggagata cagcctaata tccgacaaac tgttttacag atttacgatc gtacttgtta
     4201 cccatcattg aattttgaac atccgaacct gggagttttc cctgaaacag atagtatatt
     4261 tgaacctgta taataatata tagtctagcg ctttacggaa gagaatgtat gtatttcggt
     4321 tcctggagaa actattgcat ctattgcata ggtaatcttg cacgtcgcat ccccggttca
     4381 ttttctgcgt ttccatcttg cacttcaata gcatatcttt gttaacgaag catctgtgct
     4441 tcattttgta gaacaaaaat gcaacgcgag agcgctaatt tttcaaacaa agaatctgag
     4501 ctgcattttt acagaacaga aatgcaacgc gaaagcgcta ttttaccaac gaagaatctg
     4561 tgcttcattt ttgtaaaaca aaaatgcaac gcgagagcgc taatttttca aacaaagaat
     4621 ctgagctgca tttttacaga acagaaatgc aacgcgagag cgctatttta ccaacaaaga
     4681 atctatactt cttttttgtt ctacaaaaat gcatcccgag agcgctattt ttctaacaaa
     4741 gcatcttaga ttactttttt tctcctttgt gcgctctata atgcagtctc ttgataactt
     4801 tttgcactgt aggtccgtta aggttagaag aaggctactt tggtgtctat tttctcttcc
     4861 ataaaaaaag cctgactcca cttcccgcgt ttactgatta ctagcgaagc tgcgggtgca
     4921 ttttttcaag ataaaggcat ccccgattat attctatacc gatgtggatt gcgcatactt
     4981 tgtgaacaga aagtgatagc gttgatgatt cttcattggt cagaaaatta tgaacggttt
     5041 cttctatttt gtctctatat actacgtata ggaaatgttt acattttcgt attgttttcg
     5101 attcactcta tgaatagttc ttactacaat ttttttgtct aaagagtaat actagagata
     5161 aacataaaaa atgtagaggt cgagtttaga tgcaagttca aggagcgaaa ggtggatggg
     5221 taggttatat agggatatag cacagagata tatagcaaag agatactttt gagcaatgtt
     5281 tgtggaagcg gtattcgcaa tgccgagtgc gcggaacccc tatttgttta tttttctaaa
     5341 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     5401 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     5461 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     5521 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     5581 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     5641 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     5701 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     5761 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     5821 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     5881 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     5941 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     6001 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     6061 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     6121 gtgagcgtgg ttctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     6181 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     6241 ctgagatagg tgcctcactg attaagcatt ggtaatttcg cgtatccaac ggctgcccag
     6301 gcagccgttt ttattcctgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     6361 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     6421 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     6481 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     6541 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     6601 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     6661 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     6721 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     6781 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     6841 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     6901 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     6961 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     7021 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     7081 cagcaacgcg gcctttttac ggttcctgcc tttttcaatg ggtaataact gatataatta
     7141 aattgaagct ctaatttgtg agtttagtat acatgcattt acttataata cagtttttta
     7201 gttttgctgg ccgcatcttc tcaaatatgc ttcccagcct gcttttctgt aacgttcacc
     7261 ctctacctta gcatcccttc cctttgcaaa tagtcctctt ccaacaataa taatgtcaga
     7321 tcctgtagat accacatcat ccacggttct atactgttga cccaatgcgt cacccttgtc
     7381 atctaaaccc actccgggtg tcataatcaa ccaatcgtaa ccttcatctc ttccacccat
     7441 gtctctttga gcaataaagc cgataacaaa atctttgtcg ctcttggcaa tgtcaacagt
     7501 acccttagta tattctccag tagataggga gcccttgcat gacaattctg ctaacatcaa
     7561 aaggcctcta ggttcctttg ttacttcttc tgccgcctgc ttcaaaccgc taacaatacc
     7621 tgggcccacg acaccgtgag cattcgtaat gtctgcccat tctgctattc tgtatacacc
     7681 cgcagagtac tgcaatttga ctgtattacc aatgtcagca aattttctgt cctcgaagag
     7741 taaaaaattg tacttggcgg ataatgcctt tagcggctta actgtgccct ccatggaaaa
     7801 atcagtcaag atatccacat gtgtttttag taaacaaatt ttgggaccta atgcttcaac
     7861 taactccagt aattccttgg tggtacgaac atccaatgaa gcacacaagt ttgtttgctt
     7921 ttcgtgcatg atattaaata gcttggcagc aacaggacta ggatgagtag cagcacgttc
     7981 cttatatgta gctttcgaca tgatttatct tcgtttcctg caggtttttg ttctgtgcag
     8041 ttgggttaag aatactgggc aatttcatgt ttcttcaaca ctacatatgc gtatatatac
     8101 caatctaagt ctgtgctcct tccttcgttc ttccttctgt tcggagatta ccgaatcaaa
     8161 aaaatttcaa ggaaaccgaa atcaaaaaaa agaataaaaa aaaaatgatg aattgaattg
     8221 aaaagct
//
