LOCUS       sequence-487262-        7683 bp    DNA     circular SYN 26-SEP-2025
DEFINITION  High- copy episomal vector for galactose-inducible expression of 
            AtHKT1 protein in Saccharomyces cerevisiae..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7683)
  AUTHORS   Uozumi N, Kim EJ, Rubio F, Yamaguchi T, Muto S, Tsuboi A, Bakker EP,
            Nakamura T, Schroeder JI
  TITLE     The Arabidopsis HKT1 gene homolog mediates inward Na(+) currents in 
            xenopus laevis oocytes and Na(+) uptake in Saccharomyces cerevisiae.
  JOURNAL   Plant Physiol. 2000 Apr;122(4):1249-59. doi: 10.1104/pp.122.4.1249.
  PUBMED    10759522
REFERENCE   2  (bases 1 to 7683)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 26, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10.1104/pp.122.4"; 
            journalName: "Plant Physiol"; date: "2000-04"; volume: "122"; issue:
            "4"; pages: "1249-59"
COMMENT     Sequence Label: AtHKT1_pYES2
FEATURES             Location/Qualifiers
     source          1..7683
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(837..1084)
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     1050..1068
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(3022..3041)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3023..3041)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(3073..3514)
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     primer_bind     complement(3079..3102)
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     protein_bind    3397..3514
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     rep_origin      3536..4049
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3623..3642)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3833..3854
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      4060..4937
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     promoter        5538..5753
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             5754..6557
                     /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"
     CDS             join(6653..6721,6722..7513)
                     /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(6871..6890)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagggccg cagcttgcaa
      841 attaaagcct tcgagcgtcc caaaaccttc tcaagcaagg ttttcagtat aatgttacat
      901 gcgtacacgc gtctgtacag aaaaaaaaga aaaatttgaa atataaataa cgttcttaat
      961 actaacataa ctataaaaaa ataaataggg acctagactt caggttgtct aactccttcc
     1021 ttttcggtta gagcggatgt ggggggaggg cgtgaatgta agcgtgacat aactaattac
     1081 atgatgcggc cctctagatg catgctcgag cggccgctcg agctacgtca gggttttttt
     1141 tttttttttt tttttttttt tttttttttt tttttttttt tttttttttt tttttttttt
     1201 ttttttttct atgaaaccgg gtaaaaatac attatctgca ggccgccagt gtgagtaatt
     1261 tagttttttt tttttttttc attatacgcc ttatattata tcttcataca ttgtcataat
     1321 attaacgagg attaattaac gatgatgcaa actacataca aacttattgt acatcacaca
     1381 cagaaaagga atgctaaaag tgacatacga gactaatttt tatataacat ttatatatat
     1441 atgatcgata tgttaggaag acgaggggta aagtatccat gcgcggccag atttgactgt
     1501 gaactgctta aacctaccat aaaacattac tattattagt acgaattttc ccattggact
     1561 ccatcgtcct gcaaacccat aactcgcgtc tttgcagcca ccatcgctga tgtccagacg
     1621 ccgttcacag ctgtacccgg tagtgaaacc cacgtttcca tatgcactga taacttcgag
     1681 agtgatgtta aggacgttga aatttatcgg atcacgttgt agattttgcc tttcggtgat
     1741 tgaaatgaga aagatacata tcgtcaaaaa ggaaagttgt gacacgatga gtccactctt
     1801 tttaactttc tttccatttt cggaatcatc atctcctcct tctttctcta tcgtcttttg
     1861 ttccgtcaac ggcataaata aagtgtatgg aggaagatac atcataagaa taaagagtac
     1921 caagatagct ggggaaagtg tagagaggtc tactatagtt tctccggtgt gtcgcgaatt
     1981 caccacttga aacaacgatc caaccaactt ctcgtacgaa ctcattcctt ctagagactc
     2041 agaggtccat tcaaaggcgc agaagaaaag aagctgtatt atcagaaacc ctagcaccgt
     2101 cactccaaga agaacacata gacgaaccga gagtagatga gagtatccca tcttattgtg
     2161 gttcttgaga atgtaaccat actcgtcacg ctttgtgatc ttataaagtc cccatatgag
     2221 caaaaccaag aagcaaggga acaaagtgtt tcccatcagt acttgaggga ttaggagcca
     2281 gatgagacca gagtttttgc gaaagatgat catgttctca ttcgtgggga caaatccgca
     2341 gtttgcaaac gtggaaactg ttgtgaagac ggagaaagtg agaggtgaga tttctttgga
     2401 actaagaaca tctctcgccg ttttaacaaa atttacgtac acaagaagca acacagagcc
     2461 aactaggtta gtaacaagat ggtaactaag aaccaccgag tacaagcact tagatgccct
     2521 ttcatcgatc ttgataagac cctcacgata atcagtaaca gtctcaacgt cacagcgatc
     2581 ctcgatggaa ctgtccgaat tataagatcc caaaatatgt ctaatcttgt tatgagggaa
     2641 gacgaacttg gtgaaatagg agacgtagag gttgagaaag gaggtgaaga tttcgccacc
     2701 gaggaacatg aggatagtga ggaagataag ttgggtgttg gagaagactt ccatgtcgac
     2761 ggtagacatg gaagagacgg tgatggcaga gacagaagtg aagaaaaggt caaagtcatg
     2821 aggacgtgaa gtggttcttg gctttgtgat cttgagtgcc aaaaacccta aaaaggagaa
     2881 gaacaagaag tagatgaagt aaaggaagaa tagtgaacgt aatttagtaa gctgcgaacg
     2941 gatttttgct atttttgcca ccactctgtc cattttagtt ctcgagtaat cggatccgag
     3001 ctcggtacca agcttaatat tccctatagt gagtcgtatt acagctgcta gtagtccgat
     3061 ccggggtttt ttctccttga cgttaaagta tagaggtata ttaacaattt tttgttgata
     3121 cttttattac atttgaataa gaagtaatac aaaccgaaaa tgttgaaagt attagttaaa
     3181 gtggttatgc agtttttgca tttatatatc tgttaataga tcaaaaatca tcgcttcgct
     3241 gattaattac cccagaaata aggctaaaaa actaatcgca ttatcatcct atggttgtta
     3301 atttgattcg ttcatttgaa ggtttgtggg gccaggttac tgccaatttt tcctcttcat
     3361 aaccataaaa gctagtattg tagaatcttt attgttcgga gcagtgcggc gcgaggcaca
     3421 tctgcgtttc aggaacgcga ccggtgaaga cgaggacgca cggaggagag tcttccttcg
     3481 gagggctgtc acccgctcgg cggcttctaa tccgtggact agtggatcga tccccacgcg
     3541 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     3601 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     3661 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     3721 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     3781 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     3841 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     3901 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     3961 aattttaaca aaatattaac gcttacaatt taaatatttg cttatacaat cttcctgttt
     4021 ttggggcttt tctgattatc aaccggggtg gagcttccca ttgcgaatac cgcttccaca
     4081 aacattgctc aaaagtatct ctttgctata tatctctgtg ctatatccct atataaccta
     4141 cccatccacc tttcgctcct tgaacttgca tctaaactcg acctctacat tttttatgtt
     4201 tatctctagt attactcttt agacaaaaaa attgtagtaa gaactattca tagagtgaat
     4261 cgaaaacaat acgaaaatgt aaacatttcc tatacgtagt atatagagac aaaatagaag
     4321 aaaccgttca taattttctg accaatgaag aatcatcaac gctatcactt tctgttcaca
     4381 aagtatgcgc aatccacatc ggtatagaat ataatcgggg atgcctttat cttgaaaaaa
     4441 tgcacccgca gcttcgctag taatcagtaa acgcgggaag tggagtcagg ctttttttat
     4501 ggaagagaaa atagacacca aagtagcctt cttctaacct taacggacct acagtgcaaa
     4561 aagttatcaa gagactgcat tatagagcgc acaaaggaga aaaaaagtaa tctaagatgc
     4621 tttgttagaa aaatagcgct ctcgggatgc atttttgtag aacaaaaaag aagtatagat
     4681 tctttgttgg taaaatagcg ctctcgcgtt gcatttctgt tctgtaaaaa tgcagctcag
     4741 attctttgtt tgaaaaatta gcgctctcgc gttgcatttt tgttttacaa aaatgaagca
     4801 cagattcttc gttggtaaaa tagcgctttc gcgttgcatt tctgttctgt aaaaatgcag
     4861 ctcagattct ttgtttgaaa aattagcgct ctcgcgttgc atttttgttc tacaaaatga
     4921 agcacagatg cttcgttaac aaagatatgc tattgaagtg caagatggaa acgcagaaaa
     4981 tgaaccgggg atgcgacgtg caagattacc tatgcaatag atgcaatagt ttctccagga
     5041 accgaaatac atacattgtc ttccgtaaag cgctagacta tatattatta tacaggttca
     5101 aatatactat ctgtttcagg gaaaactccc aggttcggat gttcaaaatt caatgatggg
     5161 taacaagtac gatcgtaaat ctgtaaaaca gtttgtcgga tattaggctg tatctcctca
     5221 aagcgtattc gaatatcatt gagaagctgc agcgtcacat cggataataa tgatggcagc
     5281 cattgtagaa gtgccttttg catttctagt ctctttctcg gtctagctag ttttactaca
     5341 tcgcgaagat agaatcttag atcacactgc ctttgctgag ctggatcaat agagtaacaa
     5401 aagagtggta aggcctcgtt aaaggacaag gacctgagcg gaagtgtatc gtacagtaga
     5461 cggagtatac taggtatagt ctatagtccg tggaattaat tctcatgttt gacagcttat
     5521 catcgataag ctagcttttc aattcatcat ttttttttta ttcttttttt tgatttcggt
     5581 ttccttgaaa tttttttgat tcggtaatct ccgaacagaa ggaagaacga aggaaggagc
     5641 acagacttag attggtatat atacgcatat gtagtgttga agaaacatga aattgcccag
     5701 tattcttaac ccaactgcac agaacaaaaa cctgcaggaa acgaagataa atcatgtcga
     5761 aagctacata taaggaacgt gctgctactc atcctagtcc tgttgctgcc aagctattta
     5821 atatcatgca cgaaaagcaa acaaacttgt gtgcttcatt ggatgttcgt accaccaagg
     5881 aattactgga gttagttgaa gcattaggtc ccaaaatttg tttactaaaa acacatgtgg
     5941 atatcttgac tgatttttcc atggagggca cagttaagcc gctaaaggca ttatccgcca
     6001 agtacaattt tttactcttc gaagacagaa aatttgctga cattggtaat acagtcaaat
     6061 tgcagtactc tgcgggtgta tacagaatag cagaatgggc agacattacg aatgcacacg
     6121 gtgtggtggg cccaggtatt gttagcggtt tgaagcaggc ggcagaagaa gtaacaaagg
     6181 aacctagagg ccttttgatg ttagcagaat tgtcatgcaa gggctcccta tctactggag
     6241 aatatactaa gggtactgtt gacattgcga agagcgacaa agattttgtt atcggcttta
     6301 ttgctcaaag agacatgggt ggaagagatg aaggttacga ttggttgatt atgacacccg
     6361 gtgtgggttt agatgacaag ggagacgcat tgggtcaaca gtatagaacc gtggatgatg
     6421 tggtctctac aggatctgac attattattg ttggaagagg actatttgca aagggaaggg
     6481 atgctaaggt agagggtgaa cgttacagaa aagcaggctg ggaagcatat ttgagaagat
     6541 gcggccagca aaactaaaaa actgtattat aagtaaatgc atgtatacta aactcacaaa
     6601 ttagagcttc aatttaatta tatcagttat tacccattga aaaaggaaga gtatgagtat
     6661 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     6721 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     6781 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     6841 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     6901 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     6961 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     7021 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     7081 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     7141 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     7201 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     7261 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     7321 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     7381 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     7441 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     7501 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     7561 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     7621 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     7681 ttc
//
