LOCUS       sequence-328861-        7330 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human equilibrative nucleoside transporter 1 (hENT1).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7330)
  AUTHORS   Tominaga M, Nozaki K, Umeno D, Ishii J, Kondo A
  TITLE     Robust and flexible platform for directed evolution of yeast genetic
            switches.
  JOURNAL   Nat Commun. 2021 Mar 23;12(1):1846. doi: 10.1038/s41467-021-22134-y.
  PUBMED    33758180
REFERENCE   2  (bases 1 to 7330)
  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: "Nat 
            Commun."; date: "2021-03-23"; volume: "12(1):1846. doi"; pages: " 
            10.1038/s41467-021-22134-y"
COMMENT     Sequence Label: pGK401red-hENT1
FEATURES             Location/Qualifiers
     source          1..7330
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        897..1247
                     /gene="S. cerevisiae MET17"
                     /gene_synonym="MET15; MET25"
                     /label=MET17 promoter
                     /note="expression is repressed by methionine"
     CDS             1248..2582
                     /codon_start=1
                     /gene="S. cerevisiae MET17"
                     /gene_synonym="MET15; MET25"
                     /product="O-acetylserine and O-acetylhomoserine 
                     sulfhydrylase, required for sulfur amino acid synthesis"
                     /label=MET17
                     /translation="MPSHFDTVQLHAGQENPGDNAHRSRAVPIYATTSYVFENSKHGSQ
                     LFGLEVPGYVYSRFQNPTSNVLEERIAALEGGAAALAVSSGQAAQTLAIQGLAHTGDNI
                     VSTSYLYGGTYNQFKISFKRFGIEARFVEGDNPEEFEKVFDERTKAVYLETIGNPKYNV
                     PDFEKIVAIAHKHGIPVVVDNTFGAGGYFCQPIKYGADIVTHSATKWIGGHGTTIGGII
                     VDSGKFPWKDYPEKFPQFSQPAEGYHGTIYNEAYGNLAYIVHVRTELLRDLGPLMNPFA
                     SFLLLQGVETLSLRAERHGENALKLAKWLEQSPYVSWVSYPGLASHSHHENAKKYLSNG
                     FGGVLSFGVKDLPNADKETDPFKLSGAQVVDNLKLASNLANVGDAKTLVIAPYFTTHKQ
                     LNDKEKLASGVTKDLIRVSVGIEFIDDIIADFQQSFETVFAGQKP"
     terminator      4822..5092
                     /gene="S. cerevisae PGK1"
                     /label=PGK1 terminator
                     /note="transcription terminator for 3-phosphoglycerate 
                     kinase gene"
     primer_bind     complement(5139..5159)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5139..5157)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5178..5194)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5178..5194)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5191..5213)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5202..5218
                     /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)."
     promoter        complement(join(5226..5232,5233..5250,5251..5256))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5271..5292
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5409..5426)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5580..6168)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5660..5679)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6339..7130,7131..7199))
                     /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     6962..6981
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7200..7304)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cctcaaagtt actgacgaag cttatgatgg tttggttatt gttattggta gatggattca
       61 agggtttttg tcccaaaaga ataataacac tattgaaggc ttgaacttca tcagattact
      121 tgaaaaagat tcaggtaaat ctgagttctt attaagctac ggtaaggaag taaacaaaat
      181 tccacaaagc tacgaaaatt tgaagaaagg ttccactgta accagcaatg ggttgaactg
      241 ggaagttatt gaatatcacg cttaataaag gagaataaat cgttttctac tttcttctgc
      301 tgctataata agcacctatg ggatctatat agtattttta taacgataga ctttataaaa
      361 gaaaatacct aagtgaaaat ttggtgaatt ttgagataat tgttgggatt ccatttttaa
      421 taaggcaata atattaggta tgtagaatat actagaagtt ctcctcgagg atttaggaat
      481 ccataaaagg gaatctgcaa ttctacacaa ttctataaat attattatca tcgttttata
      541 tgttaatatt cattgatcct attacattat caatccttgc gtttcagctt ccactaattt
      601 agatgactat ttctcatcat ttgcgtcatc ttctaacacc gtatatgata atatactagt
      661 aacgtaaata ctagttagta gatgatagtt gatttttatt ccaacactaa gaaataattt
      721 cgccatttct tgaatgtatt taaagatatt taatgctata atagacattt aaatccaatt
      781 cttccaacat acaatgggag tttggccgag tggtttaagg cgtcagattt aggtggattt
      841 aacctctaaa atctctgata tcttcggatg caagggttcg aatcccttag ctctcattat
      901 tttttgcttt ttctcttgag gtcacatgat cgcaaaatgg caaatggcac gtgaagctgt
      961 cgatattggg gaactgtggt ggttggcaaa tgactaatta agttagtcaa ggcgccatcc
     1021 tcatgaaaac tgtgtaacat aataaccgaa gtgtcgaaaa ggtggcacct tgtccaattg
     1081 aacacgctcg atgaaaaaaa taagatatat ataaggttaa gtaaagcgtc tgttagaaag
     1141 gaagtttttc ctttttcttg ctctcttgtc ttttcatcta ctatttcctt cgtgtaatac
     1201 agggtcgtca gatacataga tacaattcta ttacccccat ccatacaatg ccatctcatt
     1261 tcgatactgt tcaactacac gccggccaag agaaccctgg tgacaatgct cacagatcca
     1321 gagctgtacc aatttacgcc accacttctt atgttttcga aaactctaag catggttcgc
     1381 aattgtttgg tctagaagtt ccaggttacg tctattcccg tttccaaaac ccaaccagta
     1441 atgttttgga agaaagaatt gctgctttag aaggtggtgc tgctgctttg gctgtttcct
     1501 ccggtcaagc cgctcaaacc cttgccatcc aaggtttggc acacactggt gacaacatcg
     1561 tttccacttc ttacttatac ggtggtactt ataaccagtt caaaatctcg ttcaaaagat
     1621 ttggtatcga ggctagattt gttgaaggtg acaatccaga agaattcgaa aaggtctttg
     1681 atgaaagaac caaggctgtt tatttggaaa ccattggtaa tccaaagtac aatgttccgg
     1741 attttgaaaa aattgttgca attgctcaca aacacggtat tccagttgtc gttgacaaca
     1801 catttggtgc cggtggttac ttctgtcagc caattaaata cggtgctgat attgtaacac
     1861 attctgctac caaatggatt ggtggtcatg gtactactat cggtggtatt attgttgact
     1921 ctggtaagtt cccatggaag gactacccag aaaagttccc tcaattctct caacctgccg
     1981 aaggatatca cggtactatc tacaatgaag cctacggtaa cttggcatac atcgttcatg
     2041 ttagaactga actattaaga gatttgggtc cattgatgaa cccatttgcc tctttcttgc
     2101 tactacaagg tgttgaaaca ttatctttga gagctgaaag acacggtgaa aatgcattga
     2161 agttagccaa atggttagaa caatccccat acgtatcttg ggtttcatac cctggtttag
     2221 catctcattc tcatcatgaa aatgctaaga agtatctatc taacggtttc ggtggtgtct
     2281 tatctttcgg tgtaaaagac ttaccaaatg ccgacaagga aactgaccca ttcaaacttt
     2341 ctggtgctca agttgttgac aatttaaagc ttgcctctaa cttggccaat gttggtgatg
     2401 ccaagacctt agtcattgct ccatacttca ctacccacaa acaattaaat gacaaagaaa
     2461 agttggcatc tggtgttacc aaggacttaa ttcgtgtctc tgttggtatc gaatttattg
     2521 atgacattat tgcagacttc cagcaatctt ttgaaactgt tttcgctggc caaaaaccat
     2581 gagtgtgcgt aatgagttgt aaaattatgt ataaacctac tttctctcac aagtactata
     2641 cttttataaa acgaacttta ttgaaatgaa tatccttttt ttcccttgtt acatgtcgtg
     2701 actcgtactt tgaacctaaa ttgttctaac atcaaagaac agtgttaatt cgcagtcgag
     2761 aagtcgagaa agatgccgat ttgggcgcga atcctttatt ttggcttcac cctcatacta
     2821 ttatcagggc cagaaaaagg aagtgtttcc ctccttcttg aattgatgtt accctcataa
     2881 agcacgtggc ctcttatcga gaaagaaatt accgtcgctc gtgatttgtt tgcaaaaaga
     2941 acaaaactga aaaaacccag acacgctcga cttcctgtct tcctattgat tgcagcttcc
     3001 aatttcgtca cacaacaagg tcctagcgac ggctcacagg ttttgtaaca agcaatcgaa
     3061 ggttctggaa tggcgggaaa gggtttagta ccacatgcta tgatgcccac tgtgatctcc
     3121 agagcaaagt tcgttcgatc gtactgttac tctctctctt tcaaacagaa ttgtccgaat
     3181 cgtgtgacaa caacagcctg ttctcacaca ctcttttctt ctaaccaagg gggtggttta
     3241 gtttagtaga acctcgtgaa acttacattt acatatatat aaacttgcat aaattggtca
     3301 atgcaagaaa tacatatttg gtcttttcta attcgtagtt tttcaagttc ttagatgctt
     3361 tctttttctc ttttttacag atcatcaagg aagtaattat ctacttttta caacaaatat
     3421 aaaacgctag cgtcgacatg acaaccagtc accagcctca ggacagatac aaagctgtct
     3481 ggcttatctt cttcatgctg ggtctgggaa cgctgctccc gtggaatttt ttcatgacgg
     3541 ccactcagta tttcacaaac cgcctggaca tgtcccagaa tgtgtccttg gtcactgctg
     3601 aactgagcaa ggacgcccag gcgtcagccg cccctgcagc acccttgcct gagcggaact
     3661 ctctcagtgc catcttcagc aatgtcgtga ccctatgtgc catgctgccc ctgctgttat
     3721 tcacctacct caactccttc ctgcatcaga ggatccccca gtccgtacgg atcctgggca
     3781 gcctggtggc catcctgctg gtgtttctga tcactgccat cctggtgaag gtgcagctgg
     3841 atgctctgcc cttctttgtc atcaccatga tcaagatcgt gctcattaat tcatttggtg
     3901 ccatcctgca gggcagcctg tttggtctgg ctggccttct gcctgccagc tacacggccc
     3961 ccatcatgag tggccagggc ctagcaggct tctttgcctc cgtggccatg atctgcgcta
     4021 ttgccagtgg ctcggagcta tcagaaagtg ccttcggcta ctttatcaca gcctgtgctg
     4081 ttatcatttt gaccatcatc tgttacctgg gcctgccccg cctggaattc taccgctact
     4141 accagcagct caagcttgaa ggacccgggg agcaggagac caagtcggac ctcattagca
     4201 aaggagagga gccaagagca ggcaaagagg aatctggagt ttcagtctcc aactctcagc
     4261 ccaccaatga aagccactct atcaaagcca tcctgaaaaa tatctcagtc ctggctttct
     4321 ctgtctgctt catcttcact atcaccattg ggatgtttcc agccgtgact gttgaggtca
     4381 agtccagcat cgcaggcagc agcacctggg aacgttactt cattcctgtg tcctgtttct
     4441 tgactttcaa tatctttgac tggttgggcc ggagcctcac agctgtattc atgtggcctg
     4501 ggaaggacag ccgctggctg ccaagcctgg tgctggcccg gctggtgttt gtgccactgc
     4561 tgctgctgtg caacattaag ccccgccgct acctgactgt ggtcttcgag cacgatgcct
     4621 ggttcatctt cttcatggct gcctttgcct tctccaacgg ctacctcgcc agcctctgca
     4681 tgtgcttcgg gcccaagaaa gtgaagccag ctgaggcagg gaccgcagga gccatcatgg
     4741 ccttcttcct gtgtctgggt ctggcactgg gggctgtttt ctccttcctg ttccgggcaa
     4801 ttgtgtaaag atctgaaata aattgaattg aattgaaatc gatagatcaa tttttttctt
     4861 ttctctttcc ccatccttta cgctaaaata atagtttatt ttattttttg aatatttttt
     4921 atttatatac gtatatatag actattattt atcttttaat gattattaag atttttatta
     4981 aaaaaaaatt cgctcctctt ttaatgcctt tatgcagttt ttttttccca ttcgatattt
     5041 ctatgttcgg gttcagcgta ttttaagttt aataactcga aaattctgcg ttcgttaaag
     5101 ctgcggccgc caccgcggtg gagctccagc ttttgttccc tttagtgagg gttaattgcg
     5161 cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     5221 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     5281 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     5341 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     5401 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     5461 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     5521 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     5581 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     5641 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     5701 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     5761 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     5821 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     5881 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     5941 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     6001 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     6061 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     6121 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     6181 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     6241 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     6301 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     6361 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     6421 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     6481 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     6541 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     6601 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     6661 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     6721 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     6781 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     6841 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     6901 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     6961 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     7021 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     7081 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     7141 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     7201 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     7261 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     7321 gtgccacctg
//
